Tuberculosis Laboratory Diagnosis Quality Assurance among Public Health Facilities in West Amhara Region, Ethiopia.

PubMed

Shiferaw, Melashu Balew; Hailu, Hiwot Amare; Fola, Abebe Alemu; Derebe, Mulatu Melese; Kebede, Aimro Tadese; Kebede, Abayneh Admas; Emiru, Manamnot Agegne; Gelaw, Zelalem Dessie

2015-01-01

Reliable smear microscopy is an important component of Directly Observed Treatment Scheme (DOTS) strategy for TB control program in countries with limited resources. Despite external quality assessment is established in Ethiopia, there is lower TB detection rate (48%) in Amhara region compared to the World Health Organization (WHO) estimate (70%). This highlights the quality of smear microscopy needs to be evaluated. Therefore, the aim of this study was to assess the quality of sputum smear microscopy performance among health center laboratories in West Amhara region, Ethiopia. A cross sectional study was conducted from July 08, 2013 to July 07, 2014. Data were collected from 201 public health center laboratories using a structured questionnaire. Slides were collected based on Lot Quality Assurance Sampling (LQAS) method and rechecked blindly by trained laboratory technologists. The data were entered into EPI info V.7 and smear quality indicators and AFB results were analyzed by SPSS version 20. Among 201 laboratories enrolled in this study, 47 (23.4%) laboratories had major errors. Forty one (20.4%) laboratories had a total of 67 false negative and 29 (14.4%) laboratories had a total of 68 false positive results. Specimen quality, smear thickness and evenness were found poor in 134 (66.7%), 133 (66.2%) and 126 (62.7%) laboratories, respectively. Unavailability of microscope lens cleaning solution (AOR: 2.90; 95% CI: 1.25-6.75; P: 0.013) and dirty smears (AOR: 2.65; 95% CI: 1.14-6.18; P: 0.024) were correlated with false negative results whereas no previous EQA participation (AOR: 3.43; 95% CI: 1. 39-8.45; P: 0.007) was associated with false positive results. The performance of health facilities for sputum smear microscopy was relatively poor in West Amhara region. Hence, strengthening the EQA program and technical support on sputum smear microscopy are recommended to ensure quality tuberculosis diagnostic service.

The Sensitivity and Specificity of Loop-Mediated Isothermal Amplification (LAMP) Assay for Tuberculosis Diagnosis in Adults with Chronic Cough in Malawi.

PubMed

Nliwasa, Marriott; MacPherson, Peter; Chisala, Palesa; Kamdolozi, Mercy; Khundi, McEwen; Kaswaswa, Kruger; Mwapasa, Mphatso; Msefula, Chisomo; Sohn, Hojoon; Flach, Clare; Corbett, Elizabeth L

2016-01-01

Current tuberculosis diagnostics lack sensitivity, and are expensive. Highly accurate, rapid and cheaper diagnostic tests are required for point of care use in low resource settings with high HIV prevalence. To investigate the sensitivity and specificity, and cost of loop-mediated isothermal amplification (LAMP) assay for tuberculosis diagnosis in adults with chronic cough compared to Xpert® MTB/RIF, fluorescence smear microscopy. Between October 2013 and March 2014, consecutive adults at a primary care clinic were screened for cough, offered HIV testing and assessed for tuberculosis using LAMP, Xpert® MTB/RIF and fluorescence smear microscopy. Sensitivity and specificity (with culture as reference standard), and costs were estimated. Of 273 adults recruited, 44.3% (121/273) were HIV-positive and 19.4% (53/273) had bacteriogically confirmed tuberculosis. The sensitivity of LAMP compared to culture was 65.0% (95% CI: 48.3% to 79.4%) with 100% (95% CI: 98.0% to 100%) specificity. The sensitivity of Xpert® MTB/RIF (77.5%, 95% CI: 61.5% to 89.2%) was similar to that of LAMP, p = 0.132. The sensitivity of concentrated fluorescence smear microscopy with routine double reading (87.5%, 95% CI: 73.2% to 95.8%) was higher than that of LAMP, p = 0.020. All three tests had high specificity. The lowest cost per test of LAMP was at batch size of 14 samples (US$ 9.98); this was lower than Xpert® MTB/RIF (US$ 13.38) but higher than fluorescence smear microscopy (US$ 0.65). The sensitivity of LAMP was similar to Xpert® MTB/RIF but lower than fluorescence smear microscopy; all three tests had high specificity. These findings support the Malawi policy that recommends a combination of fluorescence smear microscopy and Xpert® MTB/RIF prioritised for people living with HIV, already found to be smear-negative, or being considered for retreatment of tuberculosis.

The Sensitivity and Specificity of Loop-Mediated Isothermal Amplification (LAMP) Assay for Tuberculosis Diagnosis in Adults with Chronic Cough in Malawi

PubMed Central

Nliwasa, Marriott; MacPherson, Peter; Chisala, Palesa; Kamdolozi, Mercy; Khundi, McEwen; Kaswaswa, Kruger; Mwapasa, Mphatso; Msefula, Chisomo; Sohn, Hojoon; Flach, Clare; Corbett, Elizabeth L.

2016-01-01

Background Current tuberculosis diagnostics lack sensitivity, and are expensive. Highly accurate, rapid and cheaper diagnostic tests are required for point of care use in low resource settings with high HIV prevalence. Objective To investigate the sensitivity and specificity, and cost of loop-mediated isothermal amplification (LAMP) assay for tuberculosis diagnosis in adults with chronic cough compared to Xpert® MTB/RIF, fluorescence smear microscopy. Methods Between October 2013 and March 2014, consecutive adults at a primary care clinic were screened for cough, offered HIV testing and assessed for tuberculosis using LAMP, Xpert® MTB/RIF and fluorescence smear microscopy. Sensitivity and specificity (with culture as reference standard), and costs were estimated. Results Of 273 adults recruited, 44.3% (121/273) were HIV-positive and 19.4% (53/273) had bacteriogically confirmed tuberculosis. The sensitivity of LAMP compared to culture was 65.0% (95% CI: 48.3% to 79.4%) with 100% (95% CI: 98.0% to 100%) specificity. The sensitivity of Xpert® MTB/RIF (77.5%, 95% CI: 61.5% to 89.2%) was similar to that of LAMP, p = 0.132. The sensitivity of concentrated fluorescence smear microscopy with routine double reading (87.5%, 95% CI: 73.2% to 95.8%) was higher than that of LAMP, p = 0.020. All three tests had high specificity. The lowest cost per test of LAMP was at batch size of 14 samples (US$ 9.98); this was lower than Xpert® MTB/RIF (US$ 13.38) but higher than fluorescence smear microscopy (US$ 0.65). Conclusion The sensitivity of LAMP was similar to Xpert® MTB/RIF but lower than fluorescence smear microscopy; all three tests had high specificity. These findings support the Malawi policy that recommends a combination of fluorescence smear microscopy and Xpert® MTB/RIF prioritised for people living with HIV, already found to be smear-negative, or being considered for retreatment of tuberculosis. PMID:27171380

[Assessment of a rapid diagnostic test and portable fluorescent microscopy for malaria diagnosis in Cotonou (Bénin)].

PubMed

Ogouyèmi-Hounto, A; Kinde-Gazard, D; Keke, C; Gonçalves, E; Alapini, N; Adjovi, F; Adisso, L; Bossou, C; Denon, Y V; Massougbodji, A

2013-02-01

The aim of the study was to determine the accuracy of a rapid diagnostic test (SD Bioline Malaria Ag P.f/ Pan®) and fluorescent microscopy (CyScope®) in confirming presumptive malaria diagnosis in Cotonou. Thick blood smear was used as the reference technique for comparison. Testing was conducted on persons between the ages of 6 months and 70 years at two hospitals from June to October 2010. If malaria was suspected in the sample by the nurse based on clinical findings and sent to laboratory for confirmation, one thick smear, one rapid diagnostic test and one slide for the fluorescent microscopy were performed. All tests were read in hospital laboratories involved with the quality control of thick blood smear in the parasitology laboratory of National University Hospital of Cotonou. A total of 354 patients with clinical diagnosis of malaria were included. Malaria prevalence determined by thick smear, rapid diagnostic test and fluorescent microscopy was 22.8%, 25.4%, and 25.1% respectively. The sensitivity, specificity, positive and negative predictive values compared to the thick smears were 96.3, 95.6, 86.7, and 98.9% for rapid diagnostic test; and 97.5, 96.7, 89.8, and 99.27% for fluorescent microscopy. With these performances, these tests meet acceptability standards recommended by WHO for rapid tests (sensitivity > 95%). These two methods have advantages for the confirmation of malaria diagnosis in peripheral health structures that lack the resources to conduct diagnosis confirmation by the thick blood smear.

Sputum smear microscopy: evaluation of impact of training, microscope distribution, and use of external quality assessment guidelines for resource-poor settings.

PubMed

Van Rie, A; Fitzgerald, D; Kabuya, G; Van Deun, A; Tabala, M; Jarret, N; Behets, F; Bahati, E

2008-03-01

Sputum smear microscopy is the main and often only laboratory technique used for the diagnosis of tuberculosis in resource-poor countries, making quality assurance (QA) of smear microscopy an important activity. We evaluated the effects of a 5-day refresher training course for laboratory technicians and the distribution of new microscopes on the quality of smear microscopy in 13 primary health care laboratories in Kinshasa, Democratic Republic of Congo. The 2002 external QA guidelines for acid-fast bacillus smear microscopy were implemented, and blinded rechecking of the slides was performed before and 9 months after the training course and microscope distribution. We observed that the on-site checklist was highly time-consuming but could be tailored to capture frequent problems. Random blinded rechecking by the lot QA system method decreased the number of slides to be reviewed. Most laboratories needed further investigation for possible unacceptable performance, even according to the least-stringent interpretation. We conclude that the 2002 external QA guidelines are feasible for implementation in resource-poor settings, that the efficiency of external QA can be increased by selecting sample size parameters and interpretation criteria that take into account the local working conditions, and that greater attention should be paid to the provision of timely feedback and correction of the causes of substandard performance at poorly performing laboratories.

Comparative Analysis Reveals Potential Utility of Digital Microscopy in the Evaluation of Peripheral Blood Smears With Some Barriers to Implementation.

PubMed

Gomez-Gelvez, Juan C; Kryvenko, Oleksandr N; Chabot-Richards, Devon S; Foucar, Kathryn; Inamdar, Kedar V; Karner, Kristin H

2015-07-01

Evaluation of the peripheral blood smear (PBS) is an essential diagnostic test in current medical practice. We aimed to evaluate the use of digital microscopy for the examination of PBS as an option to provide expert interpretation to remote sites and in "on-call" situations. We collected 100 Wright-Giemsa-stained PBS slides representing normal and abnormal findings seen at a community-based hospital. Four hematopathologists independently evaluated the cases using conventional light and digital microscopy. When comparing digital vs light microscopy, most of the cellular features evaluated showed at least a moderate degree of agreement in at least three of the reviewers. Discrepancies in final diagnosis were identified in a minority of the cases, most of which were attributed to the poorer resolution of digital microscopy at high magnification (×400). These results support the limited use of digital microscopy for evaluation and triage of peripheral blood smears as a practical option to obtain expert opinion in locations where experienced staff is not available on site. Our results indicate that while digital microscopy is well suited for basic triage of these blood smears, limitations in quality of imaging at higher magnification as well as large file size may limit its utility in certain settings and situations. Copyright© by the American Society for Clinical Pathology.

[Automated procedures for microscopic analyses of blood smears: medical testing a MECOS-Ts2 complex].

PubMed

Pliasunova, S A; Balugian, R Sh; Khmel'nitskiĭ, K E; Medovyĭ, V S; Parpara, A A; Piatnitskiĭ, A M; Sokolinskiĭ, B Z; Dem'ianov, V L; Nikolaenko, D S

2006-10-01

The paper presents the results of medical tests of a group of computer-aided procedures for microscopic analysis by means of a MECOS-Ts2 complex (ZAO "MECOS", Russia), which have been conducted at the Republican Children's Clinical Hospital, the Research Institute of Emergency Pediatric Surgery and Traumatology, and Moscow City Clinical Hospital No. 23. Computer-aided procedures for calculating the differential count and for analyzing the morphology of red blood cells were tested on blood smears from a total of 443 patients and donors, computer-aided calculation of the count of reticulocytes was tested on 318 smears. The tests were carried out under the US standard NCCLS-H20A. Manual microscopy (443 smears) and flow blood analysis on a Coulter GEN*S (125 smears) were used as reference methods. The quality of collection of samples and laboriousness were additionally assessed. The certified MECOS-Ts2 subsystems were additionally used as reference tools. The tests indicated the advantage of computer-aided MECOS-Tsl2 complex microscopy over manual microscopy.

Do adjunct tuberculosis tests, when combined with Xpert MTB/RIF, improve accuracy and the cost of diagnosis in a resource-poor setting?

PubMed Central

Theron, Grant; Pooran, Anil; Peter, Jonny; van Zyl-Smit, Richard; Mishra, Hridesh Kumar; Meldau, Richard; Calligaro, Greg; Allwood, Brian; Sharma, Surendra Kumar; Dawson, Rod; Dheda, Keertan

2017-01-01

Information regarding the utility of adjunct diagnostic tests in combination with Xpert MTB/RIF (Cepheid, Sunnyvale, CA, USA) is limited. We hypothesised adjunct tests could enhance accuracy and/or reduce the cost of tuberculosis (TB) diagnosis prior to MTB/RIF testing, and rule-in or rule-out TB in MTB/RIF-negative individuals. We assessed the accuracy and/or laboratory-associated cost of diagnosis of smear microscopy, chest radiography (CXR) and interferon-γ release assays (IGRAs; T-SPOT-TB (Oxford Immunotec, Oxford, UK) and QuantiFERON-TB Gold In-Tube (Cellestis, Chadstone, Australia)) combined with MTB/RIF for TB in 480 patients in South Africa. When conducted prior to MTB/RIF: 1) smear microscopy followed by MTB/RIF (if smear negative) had the lowest cost of diagnosis of any strategy investigated; 2) a combination of smear microscopy, CXR (if smear negative) and MTB/RIF (if imaging compatible with active TB) did not further reduce the cost per TB case diagnosed; and 3) a normal CXR ruled out TB in 18% of patients (57 out of 324; negative predictive value (NPV) 100%). When downstream adjunct tests were applied to MTB/RIF-negative individuals, radiology ruled out TB in 24% (56 out of 234; NPV 100%), smear microscopy ruled in TB in 21% (seven out of 24) of culture-positive individuals and IGRAs were not useful in either context. In resource-poor settings, smear microscopy combined with MTB/RIF had the highest accuracy and lowest cost of diagnosis compared to either technique alone. In MTB/RIF-negative individuals, CXR has poor rule-in value but can reliably rule out TB in approximately one in four cases. These data inform upon the programmatic utility of MTB/RIF in high-burden settings. PMID:22075479

High throughput, detailed, cell-specific neuroanatomy of dendritic spines using microinjection and confocal microscopy

PubMed Central

Dumitriu, Dani; Rodriguez, Alfredo; Morrison, John H.

2012-01-01

Morphological features such as size, shape and density of dendritic spines have been shown to reflect important synaptic functional attributes and potential for plasticity. Here we describe in detail a protocol for obtaining detailed morphometric analysis of spines using microinjection of fluorescent dyes, high resolution confocal microscopy, deconvolution and image analysis using NeuronStudio. Recent technical advancements include better preservation of tissue resulting in prolonged ability to microinject, and algorithmic improvements that compensate for the residual Z-smear inherent in all optical imaging. Confocal imaging parameters were probed systematically for the identification of both optimal resolution as well as highest efficiency. When combined, our methods yield size and density measurements comparable to serial section transmission electron microscopy in a fraction of the time. An experiment containing 3 experimental groups with 8 subjects in each can take as little as one month if optimized for speed, or approximately 4 to 5 months if the highest resolution and morphometric detail is sought. PMID:21886104

Study on validity of a rapid diagnostic test kit versus light microscopy for malaria diagnosis in Ahmedabad city, India.

PubMed

Vyas, S; Puwar, B; Patel, V; Bhatt, G; Kulkarni, S; Fancy, M

2014-05-01

Light microscopy of blood smears for diagnosis of malaria in the field has several limitations, notably delays in diagnosis. This study in Ahmedabad in Gujarat State, India, evaluated the diagnostic performance of a rapid diagnostic test for malaria (SD Bioline Malaria Ag P.f/Pan) versus blood smear examination as the gold standard. All fever cases presenting at 13 urban health centres were subjected to rapid diagnostic testing and thick and thin blood smears. A total of 677 cases with fever were examined; 135 (20.0%) tested positive by rapid diagnostic test and 86 (12.7%) by blood smear. The sensitivity of the rapid diagnostic test for malaria was 98.8%, specificity was 91.5%, positive predictive value 63.0% and negative predictive value 99.8%. For detection of Plasmodium falciparum the sensitivity of rapid diagnostic test was 100% and specificity was 97.3%. The results show the acceptability of the rapid test as an alternative to light microscopy in the field setting.

The quality of sputum smear microscopy in public-private mix directly observed treatment laboratories in West Amhara region, Ethiopia.

PubMed

Manalebh, Almaw; Demissie, Meaza; Mekonnen, Daniel; Abera, Bayeh

2015-01-01

Ethiopia adopted Public-Private Mix Directly Observed Treatment Short Course Chemotherapy (PPM-DOTS) strategy for tuberculosis (TB) control program. Quality of sputum smear microscopy has paramount importance for tuberculosis control program in resource-poor countries like Ethiopia. A cross-sectional study was conducted to assess the quality of sputum smear microscopy in 37 Public-Private Mix laboratories in West Amhara, Ethiopia. The three external quality assessment methods (onsite evaluation, panel testing and blind rechecking) were employed. Onsite assessment revealed that 67.6% of PPM-DOTS laboratories were below the standard physical space (5 X 6) m2. The average monthly workload per laboratory technician was 19.5 (SD±2.9) slides with 12.8% positivity rate. The quality of Acid Fast Bacilli (AFB) staining reagents was sub-standard. The overall agreement for blind rechecking of 1,123 AFB slides was 99.4% (Kappa = 0.97). Reading of 370 AFB panel slides showed 3.5% false reading (Kappa = 0.92). Moreover, the consistency of reading scanty bacilli slides was lower (93%) compared to 1+, 2+ and 3+ bacilli. Based on blind rechecking and panel testing results, PPM-DOTS site laboratories showed good agreement with the reference laboratory. Physical space and qualities of AFB reagents would be areas of intervention to sustain the quality of sputum smear microscopy. Therefore, regular external quality assessment and provision of basic laboratory supplies for TB diagnosis would be the way forward to improve the quality of sputum smear microscopy services in PPM-DOTS laboratories.

The Quality of Sputum Smear Microscopy in Public-Private Mix Directly Observed Treatment Laboratories in West Amhara Region, Ethiopia

PubMed Central

Manalebh, Almaw; Demissie, Meaza; Mekonnen, Daniel; Abera, Bayeh

2015-01-01

Ethiopia adopted Public-Private Mix Directly Observed Treatment Short Course Chemotherapy (PPM-DOTS) strategy for tuberculosis (TB) control program. Quality of sputum smear microscopy has paramount importance for tuberculosis control program in resource-poor countries like Ethiopia. A cross-sectional study was conducted to assess the quality of sputum smear microscopy in 37 Public-Private Mix laboratories in West Amhara, Ethiopia. The three external quality assessment methods (onsite evaluation, panel testing and blind rechecking) were employed. Onsite assessment revealed that 67.6% of PPM-DOTS laboratories were below the standard physical space (5 X 6) m2. The average monthly workload per laboratory technician was 19.5 (SD±2.9) slides with 12.8% positivity rate. The quality of Acid Fast Bacilli (AFB) staining reagents was sub-standard. The overall agreement for blind rechecking of 1,123 AFB slides was 99.4% (Kappa = 0.97). Reading of 370 AFB panel slides showed 3.5% false reading (Kappa = 0.92). Moreover, the consistency of reading scanty bacilli slides was lower (93%) compared to 1+, 2+ and 3+ bacilli. Based on blind rechecking and panel testing results, PPM-DOTS site laboratories showed good agreement with the reference laboratory. Physical space and qualities of AFB reagents would be areas of intervention to sustain the quality of sputum smear microscopy. Therefore, regular external quality assessment and provision of basic laboratory supplies for TB diagnosis would be the way forward to improve the quality of sputum smear microscopy services in PPM-DOTS laboratories. PMID:25849516

Cost analysis of nucleic acid amplification for diagnosing pulmonary tuberculosis, within the context of the Brazilian Unified Health Care System.

PubMed

Pinto, Márcia; Entringer, Aline Piovezan; Steffen, Ricardo; Trajman, Anete

2015-01-01

We estimated the costs of a molecular test for Mycobacterium tuberculosis and resistance to rifampin (Xpert MTB/RIF) and of smear microscopy, within the Brazilian Sistema Único de Saúde (SUS, Unified Health Care System). In SUS laboratories in the cities of Rio de Janeiro and Manaus, we performed activity-based costing and micro-costing. The mean unit costs for Xpert MTB/RIF and smear microscopy were R$35.57 and R$14.16, respectively. The major cost drivers for Xpert MTB/RIF and smear microscopy were consumables/reagents and staff, respectively. These results might facilitate future cost-effectiveness studies and inform the decision-making process regarding the expansion of Xpert MTB/RIF use in Brazil.

Cost analysis of nucleic acid amplification for diagnosing pulmonary tuberculosis, within the context of the Brazilian Unified Health Care System

PubMed Central

Pinto, Márcia; Entringer, Aline Piovezan; Steffen, Ricardo; Trajman, Anete

2015-01-01

ABSTRACT We estimated the costs of a molecular test for Mycobacterium tuberculosis and resistance to rifampin (Xpert MTB/RIF) and of smear microscopy, within the Brazilian Sistema Único de Saúde (SUS, Unified Health Care System). In SUS laboratories in the cities of Rio de Janeiro and Manaus, we performed activity-based costing and micro-costing. The mean unit costs for Xpert MTB/RIF and smear microscopy were R$35.57 and R$14.16, respectively. The major cost drivers for Xpert MTB/RIF and smear microscopy were consumables/reagents and staff, respectively. These results might facilitate future cost-effectiveness studies and inform the decision-making process regarding the expansion of Xpert MTB/RIF use in Brazil. PMID:26785963

SIMPLE MEASURES ARE AS EFFECTIVE AS INVASIVE TECHNIQUES IN THE DIAGNOSIS OF PULMONARY TUBERCULOSIS IN MALAWI

PubMed Central

Bell, David J; Dacombe, Russell; Graham, Stephen M; Hicks, Alexander; Cohen, Danielle; Chikaonda, Tarsizio; French, Neil; Molyneux, Malcolm E; Zijlstra, Ed E; Squire, S Bertel; Gordon, Stephen B

2010-01-01

Setting Detection of smear-positive pulmonary tuberculosis (PTB) cases is vital for tuberculosis control. Methods to augment sputum collection are available but their additional benefit is uncertain in resource-limited settings. Objective To compare the diagnostic yields using five methods to obtain sputum from adults diagnosed with smear-negative PTB in Malawi. Design Self-expectorated sputum was collected under supervision for microscopy and mycobacterial culture in the study laboratory. Confirmed smear-negative patients, provided physiotherapy-assisted sputum and induced sputum followed, the next morning, by gastric washing and bronchoalveolar-lavage samples. Results 150 patients, diagnosed with smear-negative PTB by the hospital service, were screened. 39 (26%) were smear-positive from supervised self-expectorated sputum examined in the study laboratory. The remaining 111 confirmed smear-negative patients were enrolled; 89% were HIV positive. Seven additional smear-positive cases were diagnosed using the augmented sputum collection techniques. No differences were observed in the numbers of cases detected using the different methods. 44 (95.6%) of the 46 smear-positive cases could be detected from self-expectorated and physiotherapy-assisted samples Conclusions For countries like Malawi, the best use of limited resources to detect smear-positive PTB cases would be to improve the quality of self-expectorated sputum collection and microscopy. The additional diagnostic yield using bronchoalveolar-lavage after induced sputum is limited. PMID:19105886

Smear-positive pulmonary tuberculosis and AFB examination practices according to the standard checklist of WHO's tuberculosis laboratory assessment tool in three governmental hospitals, Eastern Ethiopia.

PubMed

Mekonnen, Abiyu

2014-05-13

Using the Directly Observed Treatment-Short course (DOTS) program the World Health Organization's global target was to detect 70% of new sputum-smear positive PTB cases. Smear positive PTB cases are more infectious than the smear negative cases. The TB case detection rate remains very low in Ethiopia, but there are increases in smear-negative PTB diagnosis which could be attributed to several factors including poor quality of sputum smear-microscopy. A five years retrospective record review of data between September, 2007 and August, 2012 and an in-depth assessment of AFB staining practices of sputum smear using a standard checklist were made. The proportion of smear positive cases relative to overall Acid Fast Bacilli (AFB) screened was determined over a five year period to indicate the overall prevalence and the trend. Odds ratio with 95 percent confidence interval was calculated for categorical variables using multivariate Logistic Regression model to assess the strength of association. A total of 1266 individuals' data were reviewed. The majority of the study participants were male, 704 (55.6%), and rural residents, 690 (54.5%). The overall prevalence rate of smear positive PTB was 21.6%. Age categories between 15-24 and 25-34 years were independent predictors of smear positive PTB with adjusted odds ratio of 2.246 [95% CI (1.098-4.597)] and 2.267 [95% CI (1.107-4.642)], respectively. More males were affected by PTB than females with an adjusted odds ratio of 1.426 [95% CI (1.083-1.879)]. An in-depth interview with the respective laboratory chiefs showed that quality control measures for sputum smear microscopy were used at different levels of the testing activities; however, equipment function verification as a quality control measure was not accomplished regularly in all of the study hospital laboratories. The smear positive PTB case detection rate indicated in this study is significantly lower than the countries which met the 70% target of the World Health Organization. Lack of feedback mechanisms in the External Quality Assurance schemes of sputum smear microscopy render the opportunity for improvement difficult; Serial sputum examination showed a considerable rate of positivity in the second sputum sample when compared with the others.

Evaluation of bleach-sedimentation for sterilising and concentrating Mycobacterium tuberculosis in sputum specimens

PubMed Central

2011-01-01

Background Bleach-sedimentation may improve microscopy for diagnosing tuberculosis by sterilising sputum and concentrating Mycobacterium tuberculosis. We studied gravity bleach-sedimentation effects on safety, sensitivity, speed and reliability of smear-microscopy. Methods This blinded, controlled study used sputum specimens (n = 72) from tuberculosis patients. Bleach concentrations and exposure times required to sterilise sputum (n = 31) were determined. In the light of these results, the performance of 5 gravity bleach-sedimentation techniques that sterilise sputum specimens (n = 16) were compared. The best-performing of these bleach-sedimentation techniques involved adding 1 volume of 5% bleach to 1 volume of sputum, shaking for 10-minutes, diluting in 8 volumes distilled water and sedimenting overnight before microscopy. This technique was further evaluated by comparing numbers of visible acid-fast bacilli, slide-reading speed and reliability for triplicate smears before versus after bleach-sedimentation of sputum specimens (n = 25). Triplicate smears were made to increase precision and were stained using the Ziehl-Neelsen method. Results M. tuberculosis in sputum was successfully sterilised by adding equal volumes of 15% bleach for 1-minute, 6% for 5-minutes or 3% for 20-minutes. Bleach-sedimentation significantly decreased the number of acid-fast bacilli visualised compared with conventional smears (geometric mean of acid-fast bacilli per 100 microscopy fields 166, 95%CI 68-406, versus 346, 95%CI 139-862, respectively; p = 0.02). Bleach-sedimentation diluted paucibacillary specimens less than specimens with higher concentrations of visible acid-fast bacilli (p = 0.02). Smears made from bleach-sedimented sputum were read more rapidly than conventional smears (9.6 versus 11.2 minutes, respectively, p = 0.03). Counting conventional acid-fast bacilli had high reliability (inter-observer agreement, r = 0.991) that was significantly reduced (p = 0.03) by bleach-sedimentation (to r = 0.707) because occasional strongly positive bleach-sedimented smears were misread as negative. Conclusions Gravity bleach-sedimentation improved laboratory safety by sterilising sputum but decreased the concentration of acid-fast bacilli visible on microscopy, especially for sputum specimens containing high concentrations of M. tuberculosis. Bleach-sedimentation allowed examination of more of each specimen in the time available but decreased the inter-observer reliability with which slides were read. Thus bleach-sedimentation effects vary depending upon specimen characteristics and whether microscopy was done for a specified time, or until a specified number of microscopy fields had been read. These findings provide an explanation for the contradictory results of previous studies. PMID:21985457

Xpert MTB/RIF assay for diagnosis of pulmonary tuberculosis in sputum specimens in remote health care facility.

PubMed

Geleta, Dereje Assefa; Megerssa, Yoseph Cherinet; Gudeta, Adugna Negussie; Akalu, Gizachew Taddesse; Debele, Melaku Tesfaye; Tulu, Kassu Desta

2015-10-19

Xpert MTB/RIF assay is considered as a great advance over conventional smear and culture in the diagnosis of TB and MDR-TB by simultaneously detecting M.tuberculosis and rifampicin resistance bacilli. However, very little information regarding the performance characteristics of Xpert MTB/RIF assay is available in Ethiopia. Therefore, the purpose of this study was to evaluate the performance of Xpert MTB/RIF assay compared to conventional sputum smear and culture methods for the diagnosis of pulmonary tuberculosis in remote health care facility. A paired expectorated sputum samples were obtained from 227 consecutively recruited patients with signs and symptoms suggestive of tuberculosis at Karamara hospital during December 2013 to May 2014. One of the sputum specimen was tested directly by Ziehl-Neelsen staining and Xpert MTB/RIF assay without NALC-NaOH decontamination. The other of pair of sputa specimen was cultured for isolation of TB bacilli by conventional methods. Diagnostic performance of Xpert MTB/RIF assay and AFB smear microscopy were calculated against culture as the gold standard. Overall 25.5% (58/227) samples were positive for Mycobacterium tuberculosis complex (MTBC) by MGIT and/or LJ media of which 36.2% (21/58) and 65.5% (35/58) were positive by AFB smear microscopy and Xpert MTB/RIF respectively. The sensitivity, specificity, as well as the positive and negative predictive value of Xpert MTB/RIF assay were 65.5% (95% CI: 53.3-77.7%), 96.3% (95% CI: 93.4-99.2%), 86.4% (95% CI: 76.2-96.5%), and 88.6% (95% CI: 83.9-93.3%) respectively. Eighteen of 58 (31%) cases that were smear microscopy negative, were positive by Xpert MTB/RIF assay. Although Xpert MTB/RIF assay demonstrated high sensitivity in detecting MTBC in sputum specimens compared with conventional AFB smear microscopy, it demonstrated suboptimal sensitivity in smear negative patients compared to conventional culture.

Impact of GeneXpert MTB/RIF assay on triage of respiratory isolation rooms for inpatients with presumed tuberculosis: a hypothetical trial.

PubMed

Chaisson, Lelia H; Roemer, Marguerite; Cantu, David; Haller, Barbara; Millman, Alexander J; Cattamanchi, Adithya; Davis, J Lucian

2014-11-15

Placing inpatients with presumed active pulmonary tuberculosis in respiratory isolation pending results of serial sputum acid-fast bacilli (AFB) smear microscopy is standard practice in high-income countries. However, this diagnostic strategy is slow and yields few tuberculosis diagnoses. We sought to determine if replacing microscopy with the GeneXpert MTB/RIF (Xpert) nucleic acid amplification assay could reduce testing time and usage of isolation rooms. We prospectively followed inpatients at San Francisco General Hospital undergoing tuberculosis evaluation. We performed smear microscopy and Xpert testing on concentrated sputum, and calculated diagnostic accuracy for both strategies in reference to serial sputum mycobacterial culture. We measured turnaround time for microscopy and estimated hypothetical turnaround times for Xpert on concentrated and unconcentrated sputum. We compared median and total isolation times for microscopy to those estimated for the 2 Xpert strategies. Among 139 patients with 142 admissions, median age was 54 years (interquartile range [IQR], 43-60 years); 32 (23%) patients were female, and 42 (30%) were HIV seropositive. Serial sputum smear microscopy and a single concentrated sputum Xpert had identical sensitivity (89%; 95% confidence interval [CI], 52%-100%) and similar specificity (99% [95% CI, 96%-100%] vs 100% [95% CI, 97%-100%]). A single concentrated sputum Xpert could have saved a median of 35 hours (IQR, 24-36 hours) in unnecessary isolation compared with microscopy, and a single unconcentrated sputum Xpert, 45 hours (IQR, 35-46 hours). Replacing serial sputum smear microscopy with a single sputum Xpert could eliminate most unnecessary isolation for inpatients with presumed tuberculosis, greatly benefiting patients and hospitals. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Does the practice of blood film microscopy for detection and quantification of malaria parasites in northwest Ethiopia fit the standard?

PubMed

Biadglegne, Fantahun; Belyhun, Yeshambel; Ali, Jemal; Walle, Fisha; Gudeta, Nigussu; Kassu, Afework; Mulu, Andargachew

2014-11-01

The diagnosis of malaria in clinical laboratories mainly depends on blood smear microscopy and this technique remains the most widely used in Ethiopia. Despite the importance of blood smear microscopy for patient's diagnosis and treatment, little effort has been made to precisely determine and identify sources of error in malaria smear microscopic diagnosis and quantification of parasitaemia. The main objective of the present study was to assess the laboratory practices of health care laboratories carrying out blood films microscopy. A cross sectional study was conducted in northwestern Ethiopia involving 29 health care institutes. A structured and pretested questionnaire were used to collect relevant information on the physical conditions, laboratory logistics and laboratory practices carrying out blood smear microscopy. There was inadequacy of laboratory reagents, guidelines and materials. Most of the health institutes have been practicing re-utilization of microscope slides for malaria microscopy. The technical procedure (preparing of reagents, making of blood films and staining of the slides) were found to be below the standard in 50% of the health institutes. Refresher training and quality assessment has been done only in two and six of the health institutes in the past five years, respectively. In most of the health care laboratories studied, availability of laboratory logistics and technical practices for malaria microscopy were found to be below the standard set by World Health Organization. Improving logistics access for malaria microscopy at all level of health care is important to increase accuracy of diagnosis and quantification of malaria parasites. Moreover, continued training and regular supervision of the staff and implementation of quality control program in the area is also crucial.

Implementation of LED fluorescence microscopy for diagnosis of pulmonary and HIV-associated tuberculosis in a hospital setting in Indonesia.

PubMed

Chaidir, Lidya; Parwati, Ida; Annisa, Jessi; Muhsinin, Soni; Meilana, Intan; Alisjahbana, Bachti; van Crevel, Reinout

2013-01-01

Fluorescence microscopy (FM) has not been implemented widely in TB endemic settings and little evaluation has been done in HIV-infected patients. We evaluated diagnostic performance, time and costs of FM with light-emitting diodes technology (LED-FM), compared with conventional (Zieh-Neelsen) microscopy in a hospital in Indonesia which acts as referral centre for HIV-infected patients. We included pulmonary tuberculosis suspects from the outpatient and HIV clinic. Direct and concentrated sputum smears were examined using LED-FM and ZN microscopy by two technicians who were blinded for the HIV-status and the result of the comparative test. Mean reading time per slide was recorded and cost of each slide was calculated. Mycobacteria culture served as the reference standard. Among 404 tuberculosis suspects from the outpatient clinic and 256 from the HIV clinic, mycobacteria culture was positive in 12.6% and 27%, respectively. The optimal sensitivity of LED-FM was achieved by using a threshold of ≥2 AFB/length. LED-FM had a higher sensitivity (75.5% vs. 54.9%, P<0.01) but lower specificity (90.0% vs 96.6%, P<0.01) compared to ZN microscopy. HIV was associated with a lower sensitivity but similar specificity. The average reading time using LED-FM was significantly shorter (2.23±0.78 vs 5.82±1.60 minutes, P<0.01), while costs per slide were similar. High sensitivity of LED-FM combined with shorter reading time of sputum smear slides make this method a potential alternative to ZN microscopy. Additional data on specificity are needed for effective implementation of this technique in high burden TB laboratories.

Added value of molecular assay Xpert MTB/RIF compared to sputum smear microscopy to assess the risk of tuberculosis transmission in a low-prevalence country.

PubMed

Opota, O; Senn, L; Prod'hom, G; Mazza-Stalder, J; Tissot, F; Greub, G; Jaton, K

2016-07-01

Airborne precautions are required at hospital admission for patients with suspected pulmonary tuberculosis. The isolation is maintained until 3 serially collected sputum smears are acid-fast bacilli negative, a time- and labor-intensive method with limited sensitivity and specificity, which has a great impact on patient flow management. We evaluated the possibility of replacing the result of microscopy by the semiquantitative result of the molecular point-of-care test Xpert MTB/RIF to assess patients' transmission risk to quickly guide airborne isolation decisions in low-endemic countries. The performance of the Xpert MTB/RIF, used as a first-line test, was compared to the results of microscopy for specimens (n=242) collected from May 2010 to December 2014 in Lausanne, Switzerland. The sensitivity and specificity of Xpert MTB/RIF were 91.5% (65/71) and 99.6% (170/171), respectively, vs. 64.8% (46/71) and 94.2% (161/171) for microscopy. Samples with negative Xpert MTB/RIF were all smear negative for Mycobacterium tuberculosis (negative predictive value, 100%). The semiquantitative results of Xpert MTB/RIF-high, medium, low or very low-were found to correlate with acid-fast bacilli detection: positive predictive value of 100% (6/6), 96.5% (27/28), 52.2% (12/23) and 11.1% (1/9) respectively. Finally, when including clinical criteria, we identified 11 smear-negative but Xpert MTB/RIF-positive patients with a significant transmission potential. In conclusion, our data support the introduction of an Xpert MTB/RIF-based strategy as a replacement of smear microscopy for a faster and more accurate management of tuberculosis patients' transmission risk in a low-prevalence country. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Role of PCR method using IS6110 primer in detecting Mycobacterium tuberculosis among the clinically diagnosed childhood tuberculosis patients at an urban hospital in Dhaka, Bangladesh.

PubMed

Kabir, Senjuti; Uddin, Mohammad Khaja Mafij; Chisti, Mohammod Jobayer; Fannana, Tilka; Haque, Mohammad Enamul; Uddin, Muhammad Reaj; Banu, Sayera; Ahmed, Tahmeed

2018-03-01

Better methods are needed for the accurate detection of child tuberculosis (TB). This study compared different laboratory tests and evaluated IS6110 PCR for the detection of Mycobacterium tuberculosis (MTB) among clinically diagnosed child TB patients. A total of 102 paediatric patients (<15 years old) with clinically diagnosed TB were enrolled in this study. The patients were admitted to the icddr,b hospital in Dhaka between 2003 and 2005. Sputum/gastric lavage samples were collected for smear microscopy, culture (solid/Lowenstein-Jensen medium and liquid/MGIT), and IS6110 PCR testing. The sensitivity, specificity, and positive and negative predictive values (PPV, NPV) of smear microscopy and PCR were compared to the two culture methods. Three patients were positive on smear microscopy (2.9%). MTB was detected by conventional culture in 15.7% (16/102), liquid culture in 14% (14/100), and IS6110 PCR in 61.8% (63/102). PCR detected an additional 45 patients who were undetected with the three other tests. Compared to conventional and liquid culture, respectively, smear microscopy showed sensitivity of 18.8% and 21.4%, specificity of 100% individually, PPV of 100% individually, and NPV of 86.9% and 88.7%, whereas PCR had sensitivity of 87.5% and 92.9%, specificity of 43% individually, PPV of 22.2% and 21%, and NPV of 94.9% and 97.4%. PCR can be useful compared to smear microscopy and culture methods and is applicable as a rapid screening test for child TB. A larger scale study is required to determine its diagnostic efficacy in improving the detection of child TB in the presence and absence of severe malnutrition. Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.

Front-Loading Sputum Microscopy Services: An Opportunity to Optimise Smear-Based Case Detection of Tuberculosis in High Prevalence Countries

PubMed Central

Ramsay, Andy; Yassin, Mohammed Ahmed; Cambanis, Alexis; Hirao, Susumu; Almotawa, Ahmad; Gammo, Mohamed; Lawson, Lovett; Arbide, Izabel; Al-Aghbari, Nasher; Al-Sonboli, Najla; Sherchand, Jeevan Bahadur; Gauchan, Punita; Cuevas, Luis Eduardo

2009-01-01

Setting. Ethiopia, Nepal, Nigeria, and Yemen. Objective. To reduce the time to complete sputum microscopy. Design. Cross-sectional surveys enrolling 923 patients with chronic cough in the 4 countries and using similar protocols. Spot-morning-spot sputum specimens were collected. An additional sputum specimen (Xspot) was collected one hour after the first, and the yields of the first two or the three specimens collected as spot-morning-spot or spot-Xspot-morning were compared. Results. 216 patients had ≥ one positive smear. 210 (97%) were identified by the spot-morning-spot, and 210 (97%) were identified by the spot-Xspot-morning specimens, with 203 and 200 identified by the first 2 specimens of each approach, respectively. Neither difference was significant. Conclusions. The time to complete smear microscopy could be reduced. PMID:20309419

A scanning electron microscopy study of diseased root surfaces conditioned with EDTA gel plus Cetavlon after scaling and root planing.

PubMed

Martins Júnior, Walter; De Rossi, Andiara; Samih Georges Abi Rached, Ricardo; Rossi, Marcos Antonio

2011-01-01

In the present investigation, a scanning electron microscopy analysis was performed to evaluate the effects of the topical application of ethylenediaminetetraacetic acid (EDTA) gel associated with Cetavlon (EDTAC) in removing the smear layer and exposing collagen fibers following root surface instrumentation. Twenty-eight teeth from adult humans, single rooted and scheduled for extraction due to periodontal reasons, were selected. Each tooth was submitted to manual (scaling and root planing) instrumentation alone or combined with ultrasonic instruments, with or without etching using a 24% EDTAC gel. Following extraction, specimens were processed and examined under a scanning electron microscope. A comparative morphological semi-quantitative analysis was performed; the intensity of the smear layer and the decalcification of cementum and dentinal surfaces were graded in 12 sets using an arbitrary scale ranging from 1 (area covered by a smear layer) to 4 (no smear layer). Root debridement with hand instruments alone or combined with ultrasonic instruments resulted in a similar smear layer covering the root surfaces. The smear layer was successfully removed from the surfaces treated with EDTAC, which exhibited numerous exposed dentinal tubules and collagen fibers. This study supports the hypothesis that manual instrumentation alone or instrumentation combined with ultrasonic instrumentation is unable to remove the smear layer, whereas the subsequent topical application of EDTAC gel effectively removes the smear layer, uncovers dentinal openings and exposes collagen fibers.

Molecular identification of leishmania species using samples obtained from negative stained smears.

PubMed

Mohaghegh, Ma; Fata, A; Salehi, Gh; Berenji, F; Bazzaz, M Mousavi; Rafatpanah, H; Parian, M; Movahedi, A

2013-04-01

Cutaneous Leishmaniasis (CL) is a parasitic skin disease. Diagnosis primarily is based on clinical signs and microscopic observation of parasite on direct stained smears or tissue sections. Sensitivity of direct smear is not as high as molecular methods. The aim of this study was to identify and characterize Leishmania species among the negative direct smears obtained from skin ulcers suspected to CL by PCR method. Among 81 patients with suspicious skin lesions to CL referred to the Parasitology lab, negative Giemsa stained smears were collected. DNA extraction performed by scraping stained smears, then PCR was performed. Among the DNA extracted from smears, L. tropica was isolated from 9 (11.1%) of the smears and L.major was not isolated from any samples. Direct microscopy on stained smears for diagnosis of leishmaniasis is not enough accurate. PCR is recommended for clinically suspected lesions with negative result of direct smear.

Serological testing versus other strategies for diagnosis of active tuberculosis in India: a cost-effectiveness analysis.

PubMed

Dowdy, David W; Steingart, Karen R; Pai, Madhukar

2011-08-01

Undiagnosed and misdiagnosed tuberculosis (TB) drives the epidemic in India. Serological (antibody detection) TB tests are not recommended by any agency, but widely used in many countries, including the Indian private sector. The cost and impact of using serology compared with other diagnostic techniques is unknown. Taking a patient cohort conservatively equal to the annual number of serological tests done in India (1.5 million adults suspected of having active TB), we used decision analysis to estimate costs and effectiveness of sputum smear microscopy (US$3.62 for two smears), microscopy plus automated liquid culture (mycobacterium growth indicator tube [MGIT], US$20/test), and serological testing (anda-tb ELISA, US$20/test). Data on test accuracy and costs were obtained from published literature. We adopted the perspective of the Indian TB control sector and an analysis frame of 1 year. Our primary outcome was the incremental cost per disability-adjusted life year (DALY) averted. We performed one-way sensitivity analysis on all model parameters, with multiway sensitivity analysis on variables to which the model was most sensitive. If used instead of sputum microscopy, serology generated an estimated 14,000 more TB diagnoses, but also 121,000 more false-positive diagnoses, 102,000 fewer DALYs averted, and 32,000 more secondary TB cases than microscopy, at approximately four times the incremental cost (US$47.5 million versus US$11.9 million). When added to high-quality sputum smears, MGIT culture was estimated to avert 130,000 incremental DALYs at an incremental cost of US$213 per DALY averted. Serology was dominated by (i.e., more costly and less effective than) MGIT culture and remained less economically favorable than sputum smear or TB culture in one-way and multiway sensitivity analyses. In India, sputum smear microscopy remains the most cost-effective diagnostic test available for active TB; efforts to increase access to quality-assured microscopy should take priority. In areas where high-quality microscopy exists and resources are sufficient, MGIT culture is more cost-effective than serology as an additional diagnostic test for TB. These data informed a recently published World Health Organization policy statement against serological tests.

Rapid Diagnosis of Tuberculosis with the Xpert MTB/RIF Assay in High Burden Countries: A Cost-Effectiveness Analysis

PubMed Central

Vassall, Anna; van Kampen, Sanne; Sohn, Hojoon; Michael, Joy S.; John, K. R.; den Boon, Saskia; Davis, J. Lucian; Whitelaw, Andrew; Nicol, Mark P.; Gler, Maria Tarcela; Khaliqov, Anar; Zamudio, Carlos; Perkins, Mark D.; Boehme, Catharina C.; Cobelens, Frank

2011-01-01

Background Xpert MTB/RIF (Xpert) is a promising new rapid diagnostic technology for tuberculosis (TB) that has characteristics that suggest large-scale roll-out. However, because the test is expensive, there are concerns among TB program managers and policy makers regarding its affordability for low- and middle-income settings. Methods and Findings We estimate the impact of the introduction of Xpert on the costs and cost-effectiveness of TB care using decision analytic modelling, comparing the introduction of Xpert to a base case of smear microscopy and clinical diagnosis in India, South Africa, and Uganda. The introduction of Xpert increases TB case finding in all three settings; from 72%–85% to 95%–99% of the cohort of individuals with suspected TB, compared to the base case. Diagnostic costs (including the costs of testing all individuals with suspected TB) also increase: from US$28–US$49 to US$133–US$146 and US$137–US$151 per TB case detected when Xpert is used “in addition to” and “as a replacement of” smear microscopy, respectively. The incremental cost effectiveness ratios (ICERs) for using Xpert “in addition to” smear microscopy, compared to the base case, range from US$41–$110 per disability adjusted life year (DALY) averted. Likewise the ICERS for using Xpert “as a replacement of” smear microscopy range from US$52–$138 per DALY averted. These ICERs are below the World Health Organization (WHO) willingness to pay threshold. Conclusions Our results suggest that Xpert is a cost-effective method of TB diagnosis, compared to a base case of smear microscopy and clinical diagnosis of smear-negative TB in low- and middle-income settings where, with its ability to substantially increase case finding, it has important potential for improving TB diagnosis and control. The extent of cost-effectiveness gain to TB programmes from deploying Xpert is primarily dependent on current TB diagnostic practices. Further work is required during scale-up to validate these findings. Please see later in the article for the Editors' Summary PMID:22087078

Cost-benefit analysis of Xpert MTB/RIF for tuberculosis suspects in German hospitals.

PubMed

Diel, Roland; Nienhaus, Albert; Hillemann, Doris; Richter, Elvira

2016-02-01

Our objective was to assess the cost-benefit of enhancing or replacing the conventional sputum smear with the real-time PCR Xpert MTB/RIF method in the inpatient diagnostic schema for tuberculosis (TB).Recent data from published per-case cost studies for TB/multidrug-resistant (MDR)-TB and from comparative analyses of sputum microscopy, mycobacterial culture, Xpert MTB/RIF and drug susceptibility testing, performed at the German National Reference Center for Mycobacteria, were used. Potential cost savings of Xpert MTB/RIF, based on test accuracy and multiple cost drivers, were calculated for diagnosing TB/MDR-TB suspects from the hospital perspective.Implementing Xpert MTB/RIF as an add-on in smear-positive and smear-negative TB suspects saves on average €48.72 and €503, respectively, per admitted patient as compared with the conventional approach. In smear-positive and smear-negative MDR-TB suspects, cost savings amount to €189.56 and €515.25 per person, respectively. Full replacement of microscopy by Xpert MTB/RIF saves €449.98. In probabilistic Monte-Carlo simulation, adding Xpert MTB/RIF is less costly in 46.4% and 76.2% of smear-positive TB and MDR-TB suspects, respectively, but 100% less expensive in all smear-negative suspects. Full replacement by Xpert MTB/RIF is also consistently cost-saving.Using Xpert MTB/RIF as an add-on to and even as a replacement for sputum smear examination may significantly reduce expenditures in TB suspects. Copyright ©ERS 2016.

Polymerase Chain Reaction in the Diagnosis of Visceral Leishmaniasis Recurrence in the Setting of Negative Splenic Smears

PubMed Central

Hasnain, Golam; Basher, Ariful; Nath, Proggananda; Ghosh, Prakash; Hossain, Faria; Hossain, Shakhawat; Mondal, Dinesh

2016-01-01

This report presents two cases of visceral leishmaniasis (VL) recurrence where the microscopy of the splenic smear failed in diagnosis. However, a strong clinical suspicion compelled further evaluation by polymerase chain reaction (PCR), which validated the etiology. This short report highlights the usefulness of PCR in diagnosing cases of suspected smear-negative VL recurrence. PMID:26556834

Low cost automated whole smear microscopy screening system for detection of acid fast bacilli.

PubMed

Law, Yan Nei; Jian, Hanbin; Lo, Norman W S; Ip, Margaret; Chan, Mia Mei Yuk; Kam, Kai Man; Wu, Xiaohua

2018-01-01

In countries with high tuberculosis (TB) burden, there is urgent need for rapid, large-scale screening to detect smear-positive patients. We developed a computer-aided whole smear screening system that focuses in real-time, captures images and provides diagnostic grading, for both bright-field and fluorescence microscopy for detection of acid-fast-bacilli (AFB) from respiratory specimens. To evaluate the performance of dual-mode screening system in AFB diagnostic algorithms on concentrated smears with auramine O (AO) staining, as well as direct smears with AO and Ziehl-Neelsen (ZN) staining, using mycobacterial culture results as gold standard. Adult patient sputum samples requesting for M. tuberculosis cultures were divided into three batches for staining: direct AO-stained, direct ZN-stained and concentrated smears AO-stained. All slides were graded by an experienced microscopist, in parallel with the automated whole smear screening system. Sensitivity and specificity of a TB diagnostic algorithm in using the screening system alone, and in combination with a microscopist, were evaluated. Of 488 direct AO-stained smears, 228 were culture positive. These yielded a sensitivity of 81.6% and specificity of 74.2%. Of 334 direct smears with ZN staining, 142 were culture positive, which gave a sensitivity of 70.4% and specificity of 76.6%. Of 505 concentrated smears with AO staining, 250 were culture positive, giving a sensitivity of 86.4% and specificity of 71.0%. To further improve performance, machine grading was confirmed by manual smear grading when the number of AFBs detected fell within an uncertainty range. These combined results gave significant improvement in specificity (AO-direct:85.4%; ZN-direct:85.4%; AO-concentrated:92.5%) and slight improvement in sensitivity while requiring only limited manual workload. Our system achieved high sensitivity without substantially compromising specificity when compared to culture results. Significant improvement in specificity was obtained when uncertain results were confirmed by manual smear grading. This approach had potential to substantially reduce workload of microscopists in high burden countries.

Miniature objective lens for array digital pathology: design improvement based on clinical evaluation

NASA Astrophysics Data System (ADS)

McCall, Brian; Pierce, Mark; Graviss, Edward A.; Richards-Kortum, Rebecca R.; Tkaczyk, Tomasz S.

2016-03-01

A miniature objective designed for digital detection of Mycobacterium tuberculosis (MTB) was evaluated for diagnostic accuracy. The objective was designed for array microscopy, but fabricated and evaluated at this stage of development as a single objective. The counts and diagnoses of patient samples were directly compared for digital detection and standard microscopy. The results were found to be correlated and highly concordant. The evaluation of this lens by direct comparison to standard fluorescence sputum smear microscopy presented unique challenges and led to some new insights in the role played by the system parameters of the microscope. The design parameters and how they were developed are reviewed in light of these results. New system parameters are proposed with the goal of easing the challenges of evaluating the miniature objective and maintaining the optical performance that produced the agreeable results presented without over-optimizing. A new design is presented that meets and exceeds these criteria.

A continuous quality improvement project to improve the quality of cervical Papanicolaou smears.

PubMed

Burkman, R T; Ward, R; Balchandani, K; Kini, S

1994-09-01

To improve the quality of cervical Papanicolaou smears by continuous quality improvement techniques. The study used a Papanicolaou smear data base of over 200,000 specimens collected between June 1988 and December 1992. A team approach employing techniques such as process flow-charting, cause and effect diagrams, run charts, and a randomized trial of collection methods was used to evaluate potential causes of Papanicolaou smear reports with the notation "inadequate" or "less than optimal" due to too few or absent endocervical cells. Once a key process variable (method of collection) was identified, the proportion of Papanicolaou smears with inadequate or absent endocervical cells was determined before and after employment of a collection technique using a spatula and Cytobrush. We measured the rate of less than optimal Papanicolaou smears due to too few or absent endocervical cells. Before implementing the new collection technique fully by June 1990, the overall rate of less than optimal cervical Papanicolaou smears ranged from 20-25%; by December 1993, it had stabilized at about 10%. Continuous quality improvement can be used successfully to study a clinical process and implement change that will lead to improvement.

A Comprehensive Evaluation of Xpert MTB/RIF Assay With Bronchoalveolar Lavage Fluid as a Single Test or Combined With Conventional Assays for Diagnosis of Pulmonary Tuberculosis in China: A Two-Center Prospective Study

PubMed Central

Pan, Xiaofu; Yang, Shoufeng; Deighton, Margaret A.; Qu, Yue; Hong, Liang; Su, Feifei

2018-01-01

Introduction: The Xpert MTB/RIF is recommended by the World Health Organization as a first line rapid test for the diagnosis of pulmonary tuberculosis (TB); however, China does not routinely use this test, partially due to the lack of a sufficient number of systematic evaluations of this assay in local patients. The aims of this study were to comprehensively assess the diagnostic performance of Xpert MTB/RIF, either alone or in combination with conventional assays for the diagnosis of pulmonary TB in adult Chinese patients. Methods: Xpert MTB/RIF tests were performed in 190 adult patients with suspected pulmonary TB, using bronchoalveolar lavage fluid (BALF) as test specimens. In parallel, conventional tests were carried out using the same BALF samples. Using two different reference standards, the performance of Xpert MTB/RIF, conventional assays and their combinations were evaluated. Results: Using mycobacterial culture as the reference comparator, Xpert MTB/RIF was found to be superior to smear-microscopy in detecting Mycobacterium tuberculosis. When final diagnosis, based on clinical criteria, was employed as the reference standard, Xpert MTB/RIF showed an even higher accuracy of 72.1%, supported by a sensitivity of 61.1% and specificity of 96.6%. Xpert MTB/RIF also demonstrated a powerful capability to identify pulmonary TB cases undetected by culture or smear-microscopy. Combining smear-microscopy and Xpert MTB/RIF was found to be the most accurate early predictor for pulmonary TB. Rifampicin resistance reported by Xpert MTB/RIF slightly deviated from that by phenotypic antibiotic susceptibility testing and requires further study with a larger sample size. Conclusion: This two-center prospective study highlights the value of Xpert MTB/RIF with BALF in diagnosing pulmonary TB in adult Chinese patients. These findings might contribute to the optimization of current diagnostic algorithms for pulmonary TB in China. PMID:29593688

A Comprehensive Evaluation of Xpert MTB/RIF Assay With Bronchoalveolar Lavage Fluid as a Single Test or Combined With Conventional Assays for Diagnosis of Pulmonary Tuberculosis in China: A Two-Center Prospective Study.

PubMed

Pan, Xiaofu; Yang, Shoufeng; Deighton, Margaret A; Qu, Yue; Hong, Liang; Su, Feifei

2018-01-01

Introduction: The Xpert MTB/RIF is recommended by the World Health Organization as a first line rapid test for the diagnosis of pulmonary tuberculosis (TB); however, China does not routinely use this test, partially due to the lack of a sufficient number of systematic evaluations of this assay in local patients. The aims of this study were to comprehensively assess the diagnostic performance of Xpert MTB/RIF, either alone or in combination with conventional assays for the diagnosis of pulmonary TB in adult Chinese patients. Methods: Xpert MTB/RIF tests were performed in 190 adult patients with suspected pulmonary TB, using bronchoalveolar lavage fluid (BALF) as test specimens. In parallel, conventional tests were carried out using the same BALF samples. Using two different reference standards, the performance of Xpert MTB/RIF, conventional assays and their combinations were evaluated. Results: Using mycobacterial culture as the reference comparator, Xpert MTB/RIF was found to be superior to smear-microscopy in detecting Mycobacterium tuberculosis . When final diagnosis, based on clinical criteria, was employed as the reference standard, Xpert MTB/RIF showed an even higher accuracy of 72.1%, supported by a sensitivity of 61.1% and specificity of 96.6%. Xpert MTB/RIF also demonstrated a powerful capability to identify pulmonary TB cases undetected by culture or smear-microscopy. Combining smear-microscopy and Xpert MTB/RIF was found to be the most accurate early predictor for pulmonary TB. Rifampicin resistance reported by Xpert MTB/RIF slightly deviated from that by phenotypic antibiotic susceptibility testing and requires further study with a larger sample size. Conclusion: This two-center prospective study highlights the value of Xpert MTB/RIF with BALF in diagnosing pulmonary TB in adult Chinese patients. These findings might contribute to the optimization of current diagnostic algorithms for pulmonary TB in China.

Influence of Smear Layer on the Antimicrobial Activity of a Sodium Hypochlorite/Etidronic Acid Irrigating Solution in Infected Dentin.

PubMed

Morago, Ana; Ordinola-Zapata, Ronald; Ferrer-Luque, Carmen María; Baca, Pilar; Ruiz-Linares, Matilde; Arias-Moliz, María Teresa

2016-11-01

The aim of this study was to evaluate the influence of the smear layer on the antimicrobial activity of a 2.5% sodium hypochlorite (NaOCl)/9% etidronic acid (HEBP) irrigating solution against bacteria growing inside dentin tubules. Dentin tubules were infected with Enterococcus faecalis by centrifugation. After 5 days of incubation, the smear layer had formed in half of the samples, which were then treated with 2.5% NaOCl either alone or combined with 9% HEBP for 3 minutes. The percentage of dead cells in infected dentinal tubules was measured using confocal laser scanning microscopy and the live/dead technique. The smear layer on the surface of the root canal wall was also observed by scanning electron microscopy. Results of the percentage of dead cells were compared using parametric tests after subjecting data to the normalized Anscombe transformation. The level of significance was P < .05. In the absence of the smear layer, 2.5% NaOCl alone and combined with 9% HEBP showed high antimicrobial activity without significant differences between the 2. The smear layer reduced the antimicrobial activity of 2.5% NaOCl significantly, whereas the solution with HEBP was not affected. No dentin tubules free of the smear layer were obtained in the 2.5% NaOCl group. In the case of 2.5% NaOCl/9% HEBP, 95.40% ± 3.63% of dentin tubules were cleaned. The presence of the smear layer reduced the antimicrobial activity of 2.5% NaOCl. The combination of 2.5% NaOCl/9% HEBP exerted antimicrobial activity that was not reduced by the smear layer. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

A multisite assessment of the quantitative capabilities of the Xpert MTB/RIF assay.

PubMed

Blakemore, Robert; Nabeta, Pamela; Davidow, Amy L; Vadwai, Viral; Tahirli, Rasim; Munsamy, Vanisha; Nicol, Mark; Jones, Martin; Persing, David H; Hillemann, Doris; Ruesch-Gerdes, Sabine; Leisegang, Felicity; Zamudio, Carlos; Rodrigues, Camilla; Boehme, Catharina C; Perkins, Mark D; Alland, David

2011-11-01

The Xpert MTB/RIF is an automated molecular test for Mycobacterium tuberculosis that estimates bacterial burden by measuring the threshold-cycle (Ct) of its M. tuberculosis-specific real-time polymerase chain reaction. Bacterial burden is an important biomarker for disease severity, infection control risk, and response to therapy. Evaluate bacterial load quantitation by Xpert MTB/RIF compared with conventional quantitative methods. Xpert MTB/RIF results were compared with smear-microscopy, semiquantiative solid culture, and time-to-detection in liquid culture for 741 patients and 2,008 samples tested in a multisite clinical trial. An internal control real-time polymerase chain reaction was evaluated for its ability to identify inaccurate quantitative Xpert MTB/RIF results. Assays with an internal control Ct greater than 34 were likely to be inaccurately quantitated; this represented 15% of M. tuberculosis-positive tests. Excluding these, decreasing M. tuberculosis Ct was associated with increasing smear microscopy grade for smears of concentrated sputum pellets (r(s) = -0.77) and directly from sputum (r(s) =-0.71). A Ct cutoff of approximately 27.7 best predicted smear-positive status. The association between M. tuberculosis Ct and time-to-detection in liquid culture (r(s) = 0.68) and semiquantitative colony counts (r(s) = -0.56) was weaker than smear. Tests of paired same-patient sputum showed that high viscosity sputum samples contained ×32 more M. tuberculosis than nonviscous samples. Comparisons between the grade of the acid-fast bacilli smear and Xpert MTB/RIF quantitative data across study sites enabled us to identify a site outlier in microscopy. Xpert MTB/RIF quantitation offers a new, standardized approach to measuring bacterial burden in the sputum of patients with tuberculosis.

Factors which contributed for low quality sputum smears for the detection of acid fast bacilli (AFB) at selected health centers in Ethiopia: A quality control perspective.

PubMed

Mekonen, Ayehu; Ayele, Yeshi; Berhan, Yifru; Woldeyohannes, Desalegn; Erku, Woldaregay; Sisay, Solomon

2018-01-01

Quality of tuberculosis (TB) microscopy diagnosis is not a guarantee despite implementation of External Quality Assurance (EQA) service in all laboratories of health facilities. Hence, we aimed at evaluating the technical quality and the findings of sputum smear microscopy for acid fast bacilli (AFB) at health centers in Hararge Zone, Oromia Region, Ethiopia. A cross-sectional study was carried out between July 8, 2014 and July 7, 2015.A pre-tested structured questionnaire was used to collect data. Lot Quality Assurance Sampling (LQAS) method was put into practice for collecting all necessary sample slides. Data were analyzed by using SPSS (Statistical Package for Social Sciences) version 20 software. P-value < 0.05 was considered as statistically significant. Of the total55 health center laboratories which had been assessed during the study period, 20 (36.4%) had major technical errors; 13 (23.6%) had 15 false negative results and 17 (30.9%) had 22 false positive results. Moreover, poor specimen quality, smear size, smear thickness, staining and evenness were indicated in 40 (72.7%), 39 (70.9%), 37 (67.3%), 27(49.1%) and 37 (67.3%) of the collected samples, respectively. False negative AFB findings were significantly associated with lack of Internal Quality Control (IQC) measures (AOR (Adjusted Odds Ratio): 2.90 (95% CI (Confidence Interval): 1.25,6.75) and poor staining procedures (AOR: 2.16(95% CI: 1.01, 5.11). The qualities of AFB smear microscopy reading and smearing were low in most of the laboratories of the health centers. Therefore, it is essential to strength EQA program through building the capacity of laboratory professionals.

Investigation of OMNIgene·SPUTUM performance in delayed tuberculosis testing by smear, culture, and Xpert MTB/RIF assays in Uganda.

PubMed

Kelly-Cirino, C D; Musisi, E; Byanyima, P; Kaswabuli, S; Andama, A; Sessolo, A; Sanyu, I; Zawedde, J; Curry, P S; Huang, L

2017-06-01

OMNIgene·SPUTUM (OM-S) is a sample transport reagent designed to work with all tuberculosis diagnostics while eliminating the need for cold chain. OM-S-treated sputum samples were assayed in several tests after multiday holds. Raw sputa from 100 patients underwent direct smear microscopy, were manually split and assigned to the OM-S group [OM-S added at collection (no other processing required) and tested after 0- to 5-day holds at room temperature] or standard-of-care (SOC) group (NaOH/N-acetyl l-cysteine decontamination, all tested on day of collection). Concentrated smear microscopy, Lowenstein Jensen (LJ) culture, and mycobacteria growth indicator tube (MGIT) culture were performed. For patients with negative direct smear, a second sample was split, with SOC (raw sputum) and OM-S portions (sediment) tested in the Xpert MTB/RIF (Xpert) assay. OM-S group and SOC group results were strongly concordant on all four tests [range, 89% (MGIT)-97% (Xpert)]. OM-S MGIT, LJ, and Xpert tests were in statistical agreement with SOC MGIT as reference. OM-S specimens had lower culture contamination rates (3% vs. 10% LJ; 2% vs. 5% MGIT) but required, on average, 5.6 additional days to become MGIT-positive. The findings suggest that samples held/transported in OM-S are compatible with smear microscopy, LJ or MGIT culture, and Xpert, and perform comparably to fresh sputum samples. Larger feasibility studies are warranted. Copyright © 2017. Published by Elsevier Ltd.

Polymerase Chain Reaction in the Diagnosis of Visceral Leishmaniasis Recurrence in the Setting of Negative Splenic Smears.

PubMed

Hasnain, Golam; Basher, Ariful; Nath, Proggananda; Ghosh, Prakash; Hossain, Faria; Hossain, Shakhawat; Mondal, Dinesh

2016-01-01

This report presents two cases of visceral leishmaniasis (VL) recurrence where the microscopy of the splenic smear failed in diagnosis. However, a strong clinical suspicion compelled further evaluation by polymerase chain reaction (PCR), which validated the etiology. This short report highlights the usefulness of PCR in diagnosing cases of suspected smear-negative VL recurrence. © The American Society of Tropical Medicine and Hygiene.

[Adhesion of sealer cements to dentin with and without smear layer].

PubMed

Gettleman, B H; Messer, H H; ElDeeb, M E

1991-01-01

The influence of a smear layer on the adhesion of sealer cements to dentin was assessed in recently extracted human anterior teeth. A total of 120 samples was tested, 40 per sealer; 20 each with and without the smear layer. The teeth were split longitudinally, and the internal surfaces were ground flat. One-half of each tooth was left with the smear layer intact, while the other half had the smear removed by washing for 3 min with 17% EDTA followed by 5.25% NaOCI. Evidence of the ability to remove the smear layer was verified by scanning electron microscopy. Using a specially designed jig, the sealer was placed into a 4-mm wide x 4 mm deep well which was then set onto the tooth.

Evaluation of GeneXpert MTB/RIF for detection of Mycobacterium tuberculosis complex and rpo B gene in respiratory and non-respiratory clinical specimens at a tertiary care teaching hospital in Saudi Arabia.

PubMed

Somily, Ali M; Barry, Mazin A; Habib, Hanan A; Alotaibi, Fawzia E; Al-Zamil, Fahad A; Khan, Mohammed A; Sarwar, Mohammed S; Bakhash, Nawab D; Alrabiaah, Abdulkarim A; Shakoor, Zahid A; Senok, Abiola C

2016-12-01

To assess the performance of Xpert MTB/RIF, an automated molecular test for Mycobacterium tuberculosis (MTB) and resistance to rifampin (RIF), against smear microscopy and culture method for diagnosis of MTB infection. Methods: This is a retrospective analysis of 103 respiratory and 137 non-respiratory patient specimens suspected of tuberculosis at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia performed between April 2014 and March 2015. Each sample underwent smear microscopy, mycobacterial culture, and GeneXpert MTB/RIF test. Results: Fifteen out of 103 respiratory samples were smear and culture positive, whereas 9 out of 137 non-respiratory samples were smear positive. Out of 9 smear positive specimens, 8 were also culture positive. All 15 culture positive respiratory samples were detected by Xpert MTB/RIF (sensitivity  and positive predictive value [PPV]=100%). Similarly, all 8 culture positive non-respiratory specimens were identified by Xpert MTB/RIF (sensitivity 100%; PPV 88.8%). The Xpert MTB/RIF detected only one false positive result in 88 smear negative respiratory specimens (specificity 98.9%; negative predictive value [NPV]= 100%). All 125 smear negative non-respiratory specimens tested negative by culture and Xpert MTB/RIF (sensitivity, specificity, PPV, NPV= 100%). Conclusion: The performance of Xpert MTB/RIF was comparable to the gold standard culture method for identification of MTB in both respiratory and non-respiratory clinical specimens.

Evaluation of GeneXpert MTB/RIF for detection of Mycobacterium tuberculosis complex and rpo B gene in respiratory and non-respiratory clinical specimens at a tertiary care teaching hospital in Saudi Arabia

PubMed Central

Somily, Ali M.; Barry, Mazin A.; Habib, Hanan A.; Alotaibi, Fawzia E.; Al-Zamil, Fahad A.; Khan, Mohammed A.; Sarwar, Mohammed S.; Bakhash, Nawab D.; Alrabiaah, Abdulkarim A.; Shakoor, Zahid A.; Senok, Abiola C.

2016-01-01

Objectives To assess the performance of Xpert MTB/RIF, an automated molecular test for Mycobacterium tuberculosis (MTB) and resistance to rifampin (RIF), against smear microscopy and culture method for diagnosis of MTB infection. Methods This is a retrospective analysis of 103 respiratory and 137 non-respiratory patient specimens suspected of tuberculosis at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia performed between April 2014 and March 2015. Each sample underwent smear microscopy, mycobacterial culture, and GeneXpert MTB/RIF test. Results Fifteen out of 103 respiratory samples were smear and culture positive, whereas 9 out of 137 non-respiratory samples were smear positive. Out of 9 smear positive specimens, 8 were also culture positive. All 15 culture positive respiratory samples were detected by Xpert MTB/RIF (sensitivity and positive predictive value [PPV]=100%). Similarly, all 8 culture positive non-respiratory specimens were identified by Xpert MTB/RIF (sensitivity 100%; PPV 88.8%). The Xpert MTB/RIF detected only one false positive result in 88 smear negative respiratory specimens (specificity 98.9%; negative predictive value [NPV]= 100%). All 125 smear negative non-respiratory specimens tested negative by culture and Xpert MTB/RIF (sensitivity, specificity, PPV, NPV= 100%). Conclusion The performance of Xpert MTB/RIF was comparable to the gold standard culture method for identification of MTB in both respiratory and non-respiratory clinical specimens. PMID:27874159

Value of light microscopy to diagnose urogenital gonorrhoea: a diagnostic test study in Indonesian clinic-based and outreach sexually transmitted infections services.

PubMed

Hananta, I Putu Yuda; van Dam, Alje P; Bruisten, Sylvia Maria; van der Loeff, Maarten Franciscus Schim; Soebono, Hardyanto; Christiaan de Vries, Henry John

2017-08-11

Gonorrhoea is a common sexually transmitted disease caused by Neisseria gonorrhoeae (Ng) infection. Light microscopy of urogenital smears is used as a simple tool to diagnose urogenital gonorrhoea in many resource-limited settings. We aimed to evaluate the accuracy of light microscopy to diagnose urogenital gonorrhoea as compared with a PCR-based test. In 2014, we examined 632 male urethral and 360 endocervical smears in clinic-based and outreach settings in Jakarta, Yogyakarta and Denpasar, Indonesia. Using the detection of Ng DNA by a validated PCR as reference test, we evaluated the accuracy of two light microscopic criteria to diagnose urogenital gonorrhoea in genital smears: (1) the presence of intracellular Gram-negative diplococci (IGND) and (2) ≥5 polymorphonuclear leucocytes (PMNL)/oil-immersion field (oif) in urethral or ≥20 PMNL/oif in endocervical smears. In male urethral smears, IGND testing had a sensitivity (95% CI), specificity (95% CI) and kappa±SE of 59.0% (50.1 to 67.4), 89.4% (86.3 to 91.9) and 0.49±0.04, respectively. For PMNL count, these were 59.0% (50.1 to 67.4), 83.7% (80.2 to 86.9) and 0.40±0.04, respectively. The accuracy of IGND in the clinic-based settings (72.0% (57.5 to 83.3), 95.2% (91.8 to 97.5) and 0.68±0.06, respectively) was better than in the outreach settings (51.2% (40.0 to 62.3), 83.4% (78.2 to 87.8) and 0.35±0.06, respectively). In endocervical smears, light microscopy performed poorly regardless of the setting or symptomatology, with kappas ranging from -0.09 to 0.24. Light microscopy using IGND and PMNL criteria can be an option with moderate accuracy to diagnose urethral gonorrhoea among males in a clinic-based setting. The poor accuracy in detecting endocervical infections indicates an urgent need to implement advanced methods, such as PCR. Further investigations are needed to identify the poor diagnostic outcome in outreach services. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

Optimization of FNAC findings as a preoperative diagnostic aid for odontogenic cysts.

PubMed

Jain, Garima; Shetty, Pushparaja

2015-01-01

Fine-needle aspiration cytology (FNAC) is not a definitive preoperative diagnostic procedure done for all cases of odontogenic cysts. This is because of the inconsistent results obtained with it. This study was done to optimize FNAC findings and help in preoperative characterization of odontogenic cysts. Cystic fluid was collected and centrifuged from 50 odontogenic cysts that were planned for excision. Three smears were prepared from the cell sediment obtained after centrifugation and stained. The stained sections were examined for presence and type of epithelial cells, to formulate a preopererative diagnosis. Epithelial cells were detected in 46% cases in smear 1, 48% cases in smear 2, and 52% cases in smear 3. When all three smears from one case were studied, 86% cases showed epithelial cells for evaluation. Cystic aspirate should be centrifuged and the entire cell sediment should be examined by making multiple smears for evaluation of cystic epithelial lining cells.

Performance of the Xpert MTB/RIF assay for the diagnosis of pulmonary tuberculosis and rifampin resistance in a low-incidence, high-resource setting.

PubMed

Rice, Jason P; Seifert, Marva; Moser, Kathleen S; Rodwell, Timothy C

2017-01-01

Performance of the Xpert MTB/RIF assay, designed to simultaneously detect Mycobacterium tuberculosis complex (MTBC) and rifampin (RIF) resistance, has been well documented in low-resource settings with high TB-incidence. However, few studies have assessed its accuracy in low TB incidence settings. We evaluated the performance of Xpert MTB/RIF using clinical sputum specimens routinely collected from suspect pulmonary TB patients over a 4-year time period in San Diego County, California. Xpert MTB/RIF results were compared to acid-fast bacilli (AFB) smear microscopy, mycobacterial culture, and phenotypic drug susceptibility testing (DST). Of 751 sputum specimens, 134 (17.8%) were MTBC culture-positive and 2 (1.5%) were multidrug-resistant (MDR). For the detection of MTBC, Xpert MTB/RIF sensitivity was 89.6% (97.7% and 74.5% in smear-positive and -negative sputa, respectively) and specificity was 97.2%; while AFB smear sensitivity and specificity were 64.9% and 77.8%, respectively. Xpert MTB/RIF detected 35 of 47 smear-negative culture-positive specimens, and excluded 124 of 137 smear-positive culture-negative specimens. Xpert MTB/RIF also correctly excluded 99.2% (121/122) of nontuberculous mycobacteria (NTM) specimens, including all 33 NTM false-positives by smear microscopy. For the detection of RIF resistance, Xpert MTB/RIF sensitivity and specificity were 100% and 98.3%, respectively. Our findings demonstrate that Xpert MTB/RIF is able to accurately detect MTBC and RIF resistance in routinely collected respiratory specimens in a low TB-incidence setting, with comparable performance to that achieved in high-incidence settings; and suggest that under these conditions the assay has particular utility in detecting smear-negative TB cases, excluding smear-positive patients without MTBC disease, and differentiating MTBC from NTM.

Performance of the Xpert MTB/RIF assay for the diagnosis of pulmonary tuberculosis and rifampin resistance in a low-incidence, high-resource setting

PubMed Central

Rice, Jason P.; Moser, Kathleen S.; Rodwell, Timothy C.

2017-01-01

Performance of the Xpert MTB/RIF assay, designed to simultaneously detect Mycobacterium tuberculosis complex (MTBC) and rifampin (RIF) resistance, has been well documented in low-resource settings with high TB-incidence. However, few studies have assessed its accuracy in low TB incidence settings. We evaluated the performance of Xpert MTB/RIF using clinical sputum specimens routinely collected from suspect pulmonary TB patients over a 4-year time period in San Diego County, California. Xpert MTB/RIF results were compared to acid-fast bacilli (AFB) smear microscopy, mycobacterial culture, and phenotypic drug susceptibility testing (DST). Of 751 sputum specimens, 134 (17.8%) were MTBC culture-positive and 2 (1.5%) were multidrug-resistant (MDR). For the detection of MTBC, Xpert MTB/RIF sensitivity was 89.6% (97.7% and 74.5% in smear-positive and -negative sputa, respectively) and specificity was 97.2%; while AFB smear sensitivity and specificity were 64.9% and 77.8%, respectively. Xpert MTB/RIF detected 35 of 47 smear-negative culture-positive specimens, and excluded 124 of 137 smear-positive culture-negative specimens. Xpert MTB/RIF also correctly excluded 99.2% (121/122) of nontuberculous mycobacteria (NTM) specimens, including all 33 NTM false-positives by smear microscopy. For the detection of RIF resistance, Xpert MTB/RIF sensitivity and specificity were 100% and 98.3%, respectively. Our findings demonstrate that Xpert MTB/RIF is able to accurately detect MTBC and RIF resistance in routinely collected respiratory specimens in a low TB-incidence setting, with comparable performance to that achieved in high-incidence settings; and suggest that under these conditions the assay has particular utility in detecting smear-negative TB cases, excluding smear-positive patients without MTBC disease, and differentiating MTBC from NTM. PMID:29016684

Cost analysis of different diagnostic algorithms for pulmonary tuberculosis varying in placement of Xpert MTB/RIF.

PubMed

Chadha, V K; Sebastian, George; Kumar, P

2016-01-01

We undertook cost analysis for diagnosis of pulmonary tuberculosis (PTB) using present algorithm under Revised National Tuberculosis Control programme and using Xpert MTB/RIF (Xpert) as frontline test or in conjunction with smear microscopy and/or chest radiography. Costs were estimated for different strategies: (A) present algorithm involving sputum smear examination followed by antibiotic trial in smear negative patients, repeat smear examination (RE) if symptoms continue and chest radiography if RE negative; (B) direct Xpert; (C) smear microscopy followed by Xpert in smear negative patients; (D) radiography followed by Xpert in those having abnormal pulmonary shadows; and (E) smear examination followed by radiography among smear negative patients and Xpert in presence of abnormal pulmonary shadow. Cost to program was estimated lowest with Strategy A and highest with Strategy B. Compared to the latter, program cost reduces by 7%, 4.5%, and 17.4% by strategies C, D, and E, respectively. Cost to the group of individuals with presumptive PTB and their attendants is significantly higher for Strategy A compared to other four strategies. Among the latter, the patients' cost was minimum with Strategy B and maximum with Strategy C. Program cost per case diagnosed was lowest by Strategy A and highest by Strategy B. Patient cost per case diagnosed was highest by Strategy A and lowest by Strategy B. Using Xpert, Strategy E had the lowest program as well as overall cost per case diagnosed. Strategy E may be chosen for diagnosis of PTB. When resources would no longer be a constraint, direct Xpert would reduce costs incurred by the patients. Copyright © 2016 Tuberculosis Association of India. Published by Elsevier B.V. All rights reserved.

Xpert® MTB/RIF assay for pulmonary tuberculosis and rifampicin resistance in adults.

PubMed

Steingart, Karen R; Schiller, Ian; Horne, David J; Pai, Madhukar; Boehme, Catharina C; Dendukuri, Nandini

2014-01-21

Accurate, rapid detection of tuberculosis (TB) and TB drug resistance is critical for improving patient care and decreasing TB transmission. Xpert® MTB/RIF assay is an automated test that can detect both TB and rifampicin resistance, generally within two hours after starting the test, with minimal hands-on technical time. The World Health Organization (WHO) issued initial recommendations on Xpert® MTB/RIF in early 2011. A Cochrane Review on the diagnostic accuracy of Xpert® MTB/RIF for pulmonary TB and rifampicin resistance was published January 2013. We performed this updated Cochrane Review as part of a WHO process to develop updated guidelines on the use of the test. To assess the diagnostic accuracy of Xpert® MTB/RIF for pulmonary TB (TB detection), where Xpert® MTB/RIF was used as both an initial test replacing microscopy and an add-on test following a negative smear microscopy result.To assess the diagnostic accuracy of Xpert® MTB/RIF for rifampicin resistance detection, where Xpert® MTB/RIF was used as the initial test replacing culture-based drug susceptibility testing (DST).The populations of interest were adults presumed to have pulmonary, rifampicin-resistant or multidrug-resistant TB (MDR-TB), with or without HIV infection. The settings of interest were intermediate- and peripheral-level laboratories. The latter may be associated with primary health care facilities. We searched for publications in any language up to 7 February 2013 in the following databases: Cochrane Infectious Diseases Group Specialized Register; MEDLINE; EMBASE; ISI Web of Knowledge; MEDION; LILACS; BIOSIS; and SCOPUS. We also searched the metaRegister of Controlled Trials (mRCT) and the search portal of the WHO International Clinical Trials Registry Platform to identify ongoing trials. We included randomized controlled trials, cross-sectional studies, and cohort studies using respiratory specimens that allowed for extraction of data evaluating Xpert® MTB/RIF against the reference standard. We excluded gastric fluid specimens. The reference standard for TB was culture and for rifampicin resistance was phenotypic culture-based DST. For each study, two review authors independently extracted data using a standardized form. When possible, we extracted data for subgroups by smear and HIV status. We assessed the quality of studies using QUADAS-2 and carried out meta-analyses to estimate pooled sensitivity and specificity of Xpert® MTB/RIF separately for TB detection and rifampicin resistance detection. For TB detection, we performed the majority of analyses using a bivariate random-effects model and compared the sensitivity of Xpert® MTB/RIF and smear microscopy against culture as reference standard. For rifampicin resistance detection, we undertook univariate meta-analyses for sensitivity and specificity separately to include studies in which no rifampicin resistance was detected. We included 27 unique studies (integrating nine new studies) involving 9557 participants. Sixteen studies (59%) were performed in low- or middle-income countries. For all QUADAS-2 domains, most studies were at low risk of bias and low concern regarding applicability.As an initial test replacing smear microscopy, Xpert® MTB/RIF pooled sensitivity was 89% [95% Credible Interval (CrI) 85% to 92%] and pooled specificity 99% (95% CrI 98% to 99%), (22 studies, 8998 participants: 2953 confirmed TB, 6045 non-TB).As an add-on test following a negative smear microscopy result, Xpert®MTB/RIF pooled sensitivity was 67% (95% CrI 60% to 74%) and pooled specificity 99% (95% CrI 98% to 99%; 21 studies, 6950 participants).For smear-positive, culture-positive TB, Xpert® MTB/RIF pooled sensitivity was 98% (95% CrI 97% to 99%; 21 studies, 1936 participants).For people with HIV infection, Xpert® MTB/RIF pooled sensitivity was 79% (95% CrI 70% to 86%; 7 studies, 1789 participants), and for people without HIV infection, it was 86% (95% CrI 76% to 92%; 7 studies, 1470 participants). Comparison with smear microscopy In comparison with smear microscopy, Xpert® MTB/RIF increased TB detection among culture-confirmed cases by 23% (95% CrI 15% to 32%; 21 studies, 8880 participants).For TB detection, if pooled sensitivity estimates for Xpert® MTB/RIF and smear microscopy are applied to a hypothetical cohort of 1000 patients where 10% of those with symptoms have TB, Xpert® MTB/RIF will diagnose 88 cases and miss 12 cases, whereas sputum microscopy will diagnose 65 cases and miss 35 cases. Rifampicin resistance For rifampicin resistance detection, Xpert® MTB/RIF pooled sensitivity was 95% (95% CrI 90% to 97%; 17 studies, 555 rifampicin resistance positives) and pooled specificity was 98% (95% CrI 97% to 99%; 24 studies, 2411 rifampicin resistance negatives). Among 180 specimens with nontuberculous mycobacteria (NTM), Xpert® MTB/RIF was positive in only one specimen that grew NTM (14 studies, 2626 participants).For rifampicin resistance detection, if the pooled accuracy estimates for Xpert® MTB/RIF are applied to a hypothetical cohort of 1000 individuals where 15% of those with symptoms are rifampicin resistant, Xpert® MTB/RIF would correctly identify 143 individuals as rifampicin resistant and miss eight cases, and correctly identify 833 individuals as rifampicin susceptible and misclassify 17 individuals as resistant. Where 5% of those with symptoms are rifampicin resistant, Xpert® MTB/RIF would correctly identify 48 individuals as rifampicin resistant and miss three cases and correctly identify 931 individuals as rifampicin susceptible and misclassify 19 individuals as resistant. In adults thought to have TB, with or without HIV infection, Xpert® MTB/RIF is sensitive and specific. Compared with smear microscopy, Xpert® MTB/RIF substantially increases TB detection among culture-confirmed cases. Xpert® MTB/RIF has higher sensitivity for TB detection in smear-positive than smear-negative patients. Nonetheless, this test may be valuable as an add-on test following smear microscopy in patients previously found to be smear-negative. For rifampicin resistance detection, Xpert® MTB/RIF provides accurate results and can allow rapid initiation of MDR-TB treatment, pending results from conventional culture and DST. The tests are expensive, so current research evaluating the use of Xpert® MTB/RIF in TB programmes in high TB burden settings will help evaluate how this investment may help start treatment promptly and improve outcomes.

Commercial Serological Antibody Detection Tests for the Diagnosis of Pulmonary Tuberculosis: A Systematic Review

PubMed Central

Steingart, Karen R; Henry, Megan; Laal, Suman; Hopewell, Philip C; Ramsay, Andrew; Menzies, Dick; Cunningham, Jane; Weldingh, Karin; Pai, Madhukar

2007-01-01

Background The global tuberculosis epidemic results in nearly 2 million deaths and 9 million new cases of the disease a year. The vast majority of tuberculosis patients live in developing countries, where the diagnosis of tuberculosis relies on the identification of acid-fast bacilli on unprocessed sputum smears using conventional light microscopy. Microscopy has high specificity in tuberculosis-endemic countries, but modest sensitivity which varies among laboratories (range 20% to 80%). Moreover, the sensitivity is poor for paucibacillary disease (e.g., pediatric and HIV-associated tuberculosis). Thus, the development of rapid and accurate new diagnostic tools is imperative. Immune-based tests are potentially suitable for use in low-income countries as some test formats can be performed at the point of care without laboratory equipment. Currently, dozens of distinct commercial antibody detection tests are sold in developing countries. The question is “do they work?” Methods and Findings We conducted a systematic review to assess the accuracy of commercial antibody detection tests for the diagnosis of pulmonary tuberculosis. Studies from all countries using culture and/or microscopy smear for confirmation of pulmonary tuberculosis were eligible. Studies with fewer than 50 participants (25 patients and 25 control participants) were excluded. In a comprehensive search, we identified 68 studies. The results demonstrate that (1) overall, commercial tests vary widely in performance; (2) sensitivity is higher in smear-positive than smear-negative samples; (3) in studies of smear-positive patients, Anda-TB IgG by enzyme-linked immunosorbent assay shows limited sensitivity (range 63% to 85%) and inconsistent specificity (range 73% to 100%); (4) specificity is higher in healthy volunteers than in patients in whom tuberculosis disease is initially suspected and subsequently ruled out; and (5) there are insufficient data to determine the accuracy of most commercial tests in smear microscopy–negative patients, as well as their performance in children or persons with HIV infection. Conclusions None of the commercial tests evaluated perform well enough to replace sputum smear microscopy. Thus, these tests have little or no role in the diagnosis of pulmonary tuberculosis. Lack of methodological rigor in these studies was identified as a concern. It will be important to review the basic science literature evaluating serological tests for the diagnosis of pulmonary tuberculosis to determine whether useful antigens have been described but their potential has not been fully exploited. Activities leading to the discovery of new antigens with immunodiagnostic potential need to be intensified. PMID:17564490

A first insight into high prevalence of undiagnosed smear-negative pulmonary tuberculosis in Northern Ethiopian prisons: implications for greater investment and quality control.

PubMed

Biadglegne, Fantahun; Rodloff, Arne C; Sack, Ulrich

2014-01-01

Tuberculosis (TB) transmission in prisons poses significant risks to inmates as well as the general population. Currently, there are no data on smear-negative pulmonary TB cases in prisons and by extension no data on the impact such cases have on TB incidence. This study was designed to obtain initial data on the prevalence of smear-negative cases of TB in prisons as well as preliminary risk factor analysis for such TB cases. This cross-sectional survey was conducted in November 2013 at eight main prisons located in the state of Amhara, Ethiopia. Interviews using a structured and pretested questionnaire were done first to identify symptomatic prisoners. Three consecutive sputum samples were collected and examined using acid fast bacilli (AFB) microscopy at the point of care. All smear-negative sputum samples were taken for culture and Xpert testing. Descriptive and multivariate analysis was done using SPSS version 16. Overall the prevalence of smear-negative pulmonary TB cases in the study prisons was 8% (16/200). Using multivariate analysis, a contact history to TB patients in prison, educational level, cough and night sweating were found to be predictors of TB positivity among smear-negative pulmonary TB cases (p ≤ 0.05). In the studied prisons, high prevalence of undiagnosed TB cases using AFB microscopy was documented, which is an important public health concern that urgently needs to be addressed. Furthermore, patients with night sweating, non-productive cough, a contact history with TB patients and who are illiterate merit special attention, larger studies are warranted in the future to assess the associations more precisely. Further studies are also needed to examine TB transmission dynamics by patients with smear-negative pulmonary TB in a prison setting.

Efficacy of Two Irrigants Used with Self-Adjusting File System on Smear Layer: A Scanning Electron Microscopy Study.

PubMed

Genç Şen, Özgür; Kaya, Sadullah; Er, Özgür; Alaçam, Tayfun

2014-01-01

Mechanical instrumentation of root canals produces a smear layer that adversely affects the root canal seal. The aim of this study was to evaluate efficacy of MTAD and citric acid solutions used with self-adjusting file (SAF) system on smear layer. Twenty-three single-rooted human teeth were used for the study. Canals were instrumented manually up to a number 20 K file size. SAF was used to prepare the root canals. The following groups were studied: Group 1: MTAD + 5.25% NaOCl, Group 2: 20% citric acid + 5.25% NaOCl, and Group 3: Control (5.25% NaOCl). All roots were split longitudinally and subjected to scanning electron microscopy. The presence of smear layer in the coronal, middle, and apical thirds was evaluated using a five-score evaluation system. Kruskal-Wallis and Mann-Whitney U tests were used for statistical analysis. In the coronal third, Group 2 exhibited the best results and was statistically different froms the other groups (P < 0.05). There was not a significant difference among the three thirds of groups according to in-group comparisons (P > 0.05). The solutions used in Group 1 and 2 could effectively remove smear layer in most of the specimens. However, citric acid was more effective than MTAD in the three thirds of the canal.

Xpert® Mtb/Rif assay for pulmonary tuberculosis and rifampicin resistance in adults

PubMed Central

Steingart, Karen R; Schiller, Ian; Horne, David J; Pai, Madhukar; Boehme, Catharina C; Dendukuri, Nandini

2014-01-01

Background Accurate, rapid detection of tuberculosis (TB) and TB drug resistance is critical for improving patient care and decreasing TB transmission. Xpert® MTB/RIF assay is an automated test that can detect both TB and rifampicin resistance, generally within two hours after starting the test, with minimal hands-on technical time. The World Health Organization (WHO) issued initial recommendations on Xpert® MTB/RIF in early 2011. A Cochrane Review on the diagnostic accuracy of Xpert® MTB/RIF for pulmonary TB and rifampicin resistance was published January 2013. We performed this updated Cochrane Review as part of a WHO process to develop updated guidelines on the use of the test. Objectives To assess the diagnostic accuracy of Xpert® MTB/RIF for pulmonary TB (TB detection), where Xpert® MTB/RIF was used as both an initial test replacing microscopy and an add-on test following a negative smear microscopy result. To assess the diagnostic accuracy of Xpert® MTB/RIF for rifampicin resistance detection, where Xpert® MTB/RIF was used as the initial test replacing culture-based drug susceptibility testing (DST). The populations of interest were adults presumed to have pulmonary, rifampicin-resistant or multidrug-resistant TB (MDR-TB), with or without HIV infection. The settings of interest were intermediate- and peripheral-level laboratories. The latter may be associated with primary health care facilities. Search methods We searched for publications in any language up to 7 February 2013 in the following databases: Cochrane Infectious Diseases Group Specialized Register; MEDLINE; EMBASE; ISI Web of Knowledge; MEDION; LILACS; BIOSIS; and SCOPUS. We also searched the metaRegister of Controlled Trials (mRCT) and the search portal of the WHO International Clinical Trials Registry Platform to identify ongoing trials. Selection criteria We included randomized controlled trials, cross-sectional studies, and cohort studies using respiratory specimens that allowed for extraction of data evaluating Xpert® MTB/RIF against the reference standard. We excluded gastric fluid specimens. The reference standard for TB was culture and for rifampicin resistance was phenotypic culture-based DST. Data collection and analysis For each study, two review authors independently extracted data using a standardized form. When possible, we extracted data for subgroups by smear and HIV status. We assessed the quality of studies using QUADAS-2 and carried out meta-analyses to estimate pooled sensitivity and specificity of Xpert® MTB/RIF separately for TB detection and rifampicin resistance detection. For TB detection, we performed the majority of analyses using a bivariate random-effects model and compared the sensitivity of Xpert® MTB/RIF and smear microscopy against culture as reference standard. For rifampicin resistance detection, we undertook univariate meta-analyses for sensitivity and specificity separately to include studies in which no rifampicin resistance was detected. Main results We included 27 unique studies (integrating nine new studies) involving 9557 participants. Sixteen studies (59%) were performed in low- or middle-income countries. For all QUADAS-2 domains, most studies were at low risk of bias and low concern regarding applicability. As an initial test replacing smear microscopy, Xpert® MTB/RIF pooled sensitivity was 89% [95% Credible Interval (CrI) 85% to 92%] and pooled specificity 99% (95% CrI 98% to 99%), (22 studies, 8998 participants: 2953 confirmed TB, 6045 non-TB).As an add-on test following a negative smear microscopy result, Xpert®MTB/RIF pooled sensitivity was 67% (95% CrI 60% to 74%) and pooled specificity 99% (95% CrI 98% to 99%; 21 studies, 6950 participants). For smear-positive, culture-positive TB, Xpert® MTB/RIF pooled sensitivity was 98% (95% CrI 97% to 99%; 21 studies, 1936 participants). For people with HIV infection, Xpert® MTB/RIF pooled sensitivity was 79% (95% CrI 70% to 86%; 7 studies, 1789 participants), and for people without HIV infection, it was 86% (95% CrI 76% to 92%; 7 studies, 1470 participants). Comparison with smear microscopy In comparison with smear microscopy, Xpert® MTB/RIF increased TB detection among culture-confirmed cases by 23% (95% CrI 15% to 32%; 21 studies, 8880 participants). For TB detection, if pooled sensitivity estimates for Xpert® MTB/RIF and smear microscopy are applied to a hypothetical cohort of 1000 patients where 10% of those with symptoms have TB, Xpert® MTB/RIF will diagnose 88 cases and miss 12 cases, whereas sputum microscopy will diagnose 65 cases and miss 35 cases. Rifampicin resistance For rifampicin resistance detection, Xpert® MTB/RIF pooled sensitivity was 95% (95% CrI 90% to 97%; 17 studies, 555 rifampicin resistance positives) and pooled specificity was 98% (95% CrI 97% to 99%; 24 studies, 2411 rifampicin resistance negatives). Among 180 specimens with nontuberculous mycobacteria (NTM), Xpert® MTB/RIF was positive in only one specimen that grew NTM (14 studies, 2626 participants). For rifampicin resistance detection, if the pooled accuracy estimates for Xpert® MTB/RIF are applied to a hypothetical cohort of 1000 individuals where 15% of those with symptoms are rifampicin resistant, Xpert® MTB/RIF would correctly identify 143 individuals as rifampicin resistant and miss eight cases, and correctly identify 833 individuals as rifampicin susceptible and misclassify 17 individuals as resistant. Where 5% of those with symptoms are rifampicin resistant, Xpert® MTB/RIF would correctly identify 48 individuals as rifampicin resistant and miss three cases and correctly identify 931 individuals as rifampicin susceptible and misclassify 19 individuals as resistant. Authors' conclusions In adults thought to have TB, with or without HIV infection, Xpert® MTB/RIF is sensitive and specific. Compared with smear microscopy, Xpert® MTB/RIF substantially increases TB detection among culture-confirmed cases. Xpert® MTB/RIF has higher sensitivity for TB detection in smear-positive than smear-negative patients. Nonetheless, this test may be valuable as an add-on test following smear microscopy in patients previously found to be smear-negative. For rifampicin resistance detection, Xpert® MTB/RIF provides accurate results and can allow rapid initiation of MDR-TB treatment, pending results from conventional culture and DST. The tests are expensive, so current research evaluating the use of Xpert® MTB/RIF in TB programmes in high TB burden settings will help evaluate how this investment may help start treatment promptly and improve outcomes. PMID:24448973

Accuracy of giant African pouched rats for diagnosing tuberculosis: comparison with culture and Xpert® MTB/RIF.

PubMed

Mulder, C; Mgode, G F; Ellis, H; Valverde, E; Beyene, N; Cox, C; Reid, S E; Van't Hoog, A H; Edwards, T L

2017-11-01

Enhanced tuberculosis (TB) case finding using detection rats in Tanzania. To assess the diagnostic accuracy of detection rats compared with culture and Xpert® MTB/RIF, and to compare enhanced case-finding algorithms using rats in smear-negative presumptive TB patients. A fully paired diagnostic accuracy study in which sputum of new adult presumptive TB patients in Tanzania was tested using smear microscopy, 11 detection rats, culture and Xpert. Of 771 eligible participants, 345 (45%) were culture-positive for Mycobacterium tuberculosis, and 264 (34%) were human immunodeficiency virus (HIV) positive. The sensitivity of the detection rats was up to 75.1% (95%CI 70.1-79.5) when compared with culture, and up to 81.8% (95%CI 76.0-86.5) when compared with Xpert, which was statistically significantly higher than the sensitivity of smear microscopy. Corresponding specificity was 40.6% (95%CI 35.9-45.5) compared with culture. The accuracy of rat detection was independent of HIV status. Using rats for triage, followed by Xpert, would result in a statistically higher yield than rats followed by light-emitting diode fluorescence microscopy, whereas the number of false-positives would be significantly lower than when using Xpert alone. Although detection rats did not meet the accuracy criteria as standalone diagnostic or triage testing for presumptive TB, they have additive value as a triage test for enhanced case finding among smear-negative TB patients if more advanced diagnostics are not available.

Microscopic diagnosis of vulvovaginal candidiasis in stained vaginal smears by Dutch general practitioners.

PubMed

Engberts, Marian K; Goedbloed, Annelize F; van Haaften, Maarten; van Haaften, Mathilde; Boon, Mathilde E; Boon, Maarten E; Heintz, Peter M

2007-01-01

To determine the accuracy of the microscopic diagnosis of vulvovaginal candidiasis (presence of [pseudo] hyphae and blastospores) in stained vaginal smears in clinical practice. General practitioners trained in diagnosing vulvovaginal candidiasis performed microscopy of 324 stained vaginal smears. These smears were sent to the pathologist for confirmation of the microscopic diagnosis of the clinician; cytologic diagnosis by the pathologist was considered the gold standard. In 104 of the 342 cases Candida was established by the pathologist. The clinicians made 24 false positive and 50 false negative diagnoses of Candida. Sensitivity and specificity of the microscopic diagnoses of the clinicians were 52% and 89%, respectively. The most frequent reason for a false positive diagnosis was presence of hairs, whereas the most frequent reason for a false negative diagnosis was understaining of the smear. This study shows that even in stained smears it is difficult for clinicians to recognize blastospores and (pseudo)hyphae. Efforts are clearly needed to improve the quality of the clinical diagnosis of vulvovaginal candidiasis.

Simplified Pan-species Real-time PCR-based Detection of Plasmodium Spp. in Blood Smear.

PubMed

Hassanpour, Gholamreza; Mirhendi, Hossein; Mohebali, Mehdi; Raeisi, Ahmad; Zeraati, Hojjat; Keshavarz, Hossein

2016-01-01

We aimed to quicken and simplify the detection of Plasmodium in blood samples by developing and testing a pan- Plasmodium real-time PCR for accurate screening of individuals suspected of malaria. A single primer/probe set for pan-species Plasmodium -specific real time PCR targeting a conserved region of the small subunit 18S ribosomal DNA was designed and evaluated for rapid diagnosis and screening of malaria infections using dried blood smears. FTA cards were used for rapid and simple DNA extraction. The primers and probes showed a positive response with the DNA extracted from bloods infected with P. falciparum and P. vivax but not with DNA extracted from various smears from uninfected blood samples. Seven positive cases positive by both microscopy and nested PCR were found among 280 blood samples taken from in South and Southeast Iran. Five samples were identified as positive for P. vivax and two as positive for P. falciparum . All positive samples were positive by real-time PCR. Furthermore, all 38-blood samples positive by microscopy were positive by real-time PCR. No microscopy-negative samples were positive by real-time PCR. By using a simple FTA card for DNA extraction and by application of the real-time PCR developed in this study, sensitivity similar to nested-PCR and microscopy was achieved. This format simplifies the detection of Plasmodium in large numbers of samples.

The use of light-emitting diode fluorescence to diagnose mycobacterial lymphadenitis in fine-needle aspirates from children

PubMed Central

van Wyk, A. C.; Marais, B. J.; Warren, R. M.; van Wyk, S. S.; Wright, C. A.

2011-01-01

SUMMARY BACKGROUND Fine-needle aspiration biopsy (FNAB) is a simple, safe and effective method for investigating suspected mycobacterial lymphadenitis in children. Fluorescence microscopy can provide rapid mycobacterial confirmation. Light-emitting diodes (LEDs) provide a cheap and robust excitation light source, making fluorescence microscopy feasible in resource-limited settings. OBJECTIVE To compare the diagnostic performance of LED fluorescence microscopy on Papanicolaou (PAP) stained smears with the conventional mercury vapour lamp (MVL). METHODS FNAB smears routinely collected from palpable lymph nodes in children with suspected mycobacterial disease were PAP-stained and evaluated by two independent microscopists using different excitatory light sources (MVL and LED). Mycobacterial culture results provided the reference standard. A manually rechargeable battery-powered LED power source was evaluated in a random subset. RESULTS We evaluated 182 FNAB smears from 121 children (median age 31 months, interquartile range 10–67). Mycobacterial cultures were positive in 84 of 121 (69%) children. The mean sensitivity with LED (mains-powered), LED (rechargeable battery-powered) and MVL was respectively 48.2%, 50.0% and 51.8% (specificity 78.4%, 86.7% and 78.4%). Inter-observer variation was similar for LED and MVL (κ = 0.5). CONCLUSION LED fluorescence microscopy provides a reliable alternative to conventional methods and has many favourable attributes that would facilitate improved, decentralised diagnostic services. PMID:21276297

Evaluation of pulmonary tuberculosis case detection improvement with the deployment of XpertMTB/Rif in the tuberculosis control program of cross River State, Nigeria.

PubMed

Ochang, Ernest Afu; Emanghe, Ubleni E; Ewa, Atana; Otu, Akaninyene; Offor, Jonah B; Odo, Micheal; Etokidem, Aniekan; Afirima, Barinadaa; Owuna, Oju Eni; Obeten, Sunday M; Meremikwu, Martin M

2017-01-01

Global indices show that Nigeria has the highest tuberculosis (TB)-related mortality rate. Overdependence on Ziehl-Neelsen (ZN) smear microscopy for diagnosis and human immunodeficiency virus (HIV)/AIDS has limited control efforts. The new polymerase chain reaction-based XpertMTB/Rif (Cepheid Inc., CA, USA), which detects Mycobacterium tuberculosis and rifampicin resistance, was introduced in Cross River State in 2014. We evaluated the increment in pulmonary TB case detection following introduction of XpertMTB/Rif into the Cross River State TB control program. Data from three XpertMTB/Rif centers in Cross River were prospectively collected from June 2014 to December 2015. One spot specimen and one early morning sputum specimen were collected from each patient and tested using microscopy while one specimen was used for XpertMTB/Rif. A total of 2326 patients comprising 47.4.0% (1103) males and 52.6% (1223) females were evaluated. Their mean age was 38.8 years (range 4-89 years); 42.6% (991) were HIV positive and 50.9% (1183) HIV negative, and for 6.5% (158) HIV status was unknown. XpertMTB/Rif detected M. tuberculosis in 22.9% (534) of patients, while 16.8% (391) were ZN smear positive. Smear microscopy missed 24.5% (131/534) of cases (P < 0.0001). When patients where categorized according to HIV status, XpertMTB/Rif detected 23.7% (280/1183) and ZN smear microscopy detected 18.5% (219/1183) of HIV-negative patients. XpertMTB/Rif detected 21.5% (213/991) and ZN smear 14.1% (140/991) of HIV-positive patients. TB case detection was significantly higher in HIV-negative patients than in HIV-positive patients when either XpertMTB/Rif and/or ZN was used (P = 0.018 and 0.012, respectively). The use of XpertMTB/Rif has significantly increased TB case detection and data in Cross River State. Scale-up of additional strategies such as culture is still required to improve TB detection in HIV patients.

External quality assessment of AFB smear microscopy performances and its associated factors in selected private health facilities in Addis Ababa, Ethiopia.

PubMed

Mosissa, Lemi; Kebede, Abebaw; Mindaye, Tedla; Getahun, Muluwork; Tulu, Sisay; Desta, Kassu

2016-01-01

Tuberculosis (TB) is still a public health problem in sub Saharan African countries. In resource-limited settings, TB diagnosis relies on sputum smear microscopy, with low and variable sensitivities, especially in paucibacillary pediatric and HIV-associated TB patients. Tuberculosis microscopy centers have several weaknesses like overworking, insufficiently trained personnel, inconsistent reagent supplies, and poorly maintained equipments; thus, there is a critical need for investments in laboratory infrastructure, capacity building, and quality assurance schemes. The performance of TB microscopy centers in the private health facilities in Addis Ababa is not known so far. The main objective of the study was to assess laboratory performance of acid fast bacilli (AFB) smear microscopy and its associated factors in selected private health facilities in Addis Ababa, Ethiopia. A cross-sectional study was conducted in 33 selected private health facilities of Addis Ababa, Ethiopia comprising 7 hospitals, 2 NGO health centers, 23 higher clinics and 1 diagnostic laboratory that provide AFB smear microscopy services. The study was conducted from January to April 2014. A total of 283 stained sputum smears were randomly collected from participant laboratories for blinded rechecking, 320 panel slides were sent to 32 microscopy centers to evaluate their performance on AFB reading, staining and reporting. Checklists were used to assess quality issues of laboratories. Data were captured, cleaned, and analyzed using SPSS version 16.0; χ(2) tests, kappa statistics were used for comparison purpose. P value < 0.05 considered statistically significant. Among the 32 participant laboratories, 2-scored 100%, 15 scored 80-95% & the remaining 15 scored 50-75% for overall proficiency test performance. There were 10 (3.15%) major errors and 121 (37.8%) minor errors. The sensitivity, specificity, PPV and NPV of panel reading by microscopy centers were 89%, 96%, 96%, and 90% respectively. Out of 283 randomly selected slides for blind rechecking, 11 (3.9%) slides interpreted falsely for AFB, with overall agreement of 97.5%, sensitivity of 88.4% and specificity of 99.3%. In terms of slide quality assessment, 71.6% of AFB slides were graded as good for evenness, cleanness, thickness, size, staining and labeling. The performance score for AFB slide evenness was 56.9% (161 slides) and for labeling quality was 90.8% (257 slides); having significant difference in slide quality (p value < 0.05). On-site evaluation indicated problems in terms of infrastructure, standard operating procedure, reagent quality; equipment maintenance, data management and training issues. Most of the health facilities had poor maintenance scheme for microscope (53.5%) and poor inventory management (25.0%) system. Microscopy centers that scored a proficiency of 75.5%; which is below the acceptable minimum score of 80% and an overall error rate of 3.9% for blinded rechecking needs attention. Moreover, there are gaps identified through on site assessment including poor SOP, reagent quality, equipment maintenance, data management & lack of updated training on AFB microscopy techniques, requiring a concerted effort to alleviate the bottle neck problems and strengthening the public private partnership to control TB.

External quality assessment of AFB smear microscopy performances and its associated factors in selected private health facilities in Addis Ababa, Ethiopia

PubMed Central

Mosissa, Lemi; Kebede, Abebaw; Mindaye, Tedla; Getahun, Muluwork; Tulu, Sisay; Desta, Kassu

2016-01-01

Tuberculosis (TB) is still a public health problem in sub Saharan African countries. In resource-limited settings, TB diagnosis relies on sputum smear microscopy, with low and variable sensitivities, especially in paucibacillary pediatric and HIV-associated TB patients. Tuberculosis microscopy centers have several weaknesses like overworking, insufficiently trained personnel, inconsistent reagent supplies, and poorly maintained equipments; thus, there is a critical need for investments in laboratory infrastructure, capacity building, and quality assurance schemes. The performance of TB microscopy centers in the private health facilities in Addis Ababa is not known so far. The main objective of the study was to assess laboratory performance of acid fast bacilli (AFB) smear microscopy and its associated factors in selected private health facilities in Addis Ababa, Ethiopia. A cross-sectional study was conducted in 33 selected private health facilities of Addis Ababa, Ethiopia comprising 7 hospitals, 2 NGO health centers, 23 higher clinics and 1 diagnostic laboratory that provide AFB smear microscopy services. The study was conducted from January to April 2014. A total of 283 stained sputum smears were randomly collected from participant laboratories for blinded rechecking, 320 panel slides were sent to 32 microscopy centers to evaluate their performance on AFB reading, staining and reporting. Checklists were used to assess quality issues of laboratories. Data were captured, cleaned, and analyzed using SPSS version 16.0; χ2 tests, kappa statistics were used for comparison purpose. P value < 0.05 considered statistically significant. Among the 32 participant laboratories, 2-scored 100%, 15 scored 80-95% & the remaining 15 scored 50-75% for overall proficiency test performance. There were 10 (3.15%) major errors and 121 (37.8%) minor errors. The sensitivity, specificity, PPV and NPV of panel reading by microscopy centers were 89%, 96%, 96%, and 90% respectively. Out of 283 randomly selected slides for blind rechecking, 11 (3.9%) slides interpreted falsely for AFB, with overall agreement of 97.5%, sensitivity of 88.4% and specificity of 99.3%. In terms of slide quality assessment, 71.6% of AFB slides were graded as good for evenness, cleanness, thickness, size, staining and labeling. The performance score for AFB slide evenness was 56.9% (161 slides) and for labeling quality was 90.8% (257 slides); having significant difference in slide quality (p value < 0.05). On-site evaluation indicated problems in terms of infrastructure, standard operating procedure, reagent quality; equipment maintenance, data management and training issues. Most of the health facilities had poor maintenance scheme for microscope (53.5%) and poor inventory management (25.0%) system. Microscopy centers that scored a proficiency of 75.5%; which is below the acceptable minimum score of 80% and an overall error rate of 3.9% for blinded rechecking needs attention. Moreover, there are gaps identified through on site assessment including poor SOP, reagent quality, equipment maintenance, data management & lack of updated training on AFB microscopy techniques, requiring a concerted effort to alleviate the bottle neck problems and strengthening the public private partnership to control TB. PMID:27642463

Effects of different peracetic acid formulations on post space radicular dentin.

PubMed

Belizário, Lauriê Garcia; Kuga, Milton Carlos; Castro-Núñez, Gabriela Mariana; Escalante-Otárola, Wilfredo Gustavo; Só, Marcus Vinicius Reis; Pereira, Jefferson Ricardo

2018-01-05

The optimal irrigating solution with antimicrobial and dentin cleansing properties for post space preparation for fiber posts is unclear. Peracetic acid is one option but is available in various chemical formulations that require evaluation. The purpose of this in vitro study was to evaluate dentin surface cleanliness based on the presence of a smear layer and the number of open dentin tubules. It also investigates the chemical composition of residues after canal irrigation with a 1% peracetic acid solution (PA) at low or high concentration of hydrogen peroxide during the preparation of intracanal fiber posts. After filling the root canals of 40 mandibular incisors, a rotary instrument was used for intracanal preparation to place fiber posts. The teeth were divided into 4 groups (n=10) according to the post space irrigation protocol as follows: CG (control): distilled water; NA (NaOCl): 2.5% sodium hypochlorite; LH: PA with low concentration of hydrogen peroxide; and HH: PA with high concentrations of hydrogen peroxide. After irrigation, the teeth were sectioned, and the intracanal dentin surface was subjected to analysis using energy dispersive spectroscopy to evaluate chemical composition and to scanning electron microscopy (×500) to evaluate the presence of the smear layer. The number of open dentin tubules was measured by scanning electron microscopy analysis (×2000) using photo-editing software. ANOVA and the Tukey test (α=.05) were used to evaluate the data, except for the presence of a smear layer, for which the Kruskal-Wallis and Dunn tests were used (α=.05). The highest concentrations of oxygen in the dentin residues were detected in LH and HH (P<.05); CG and NA showed similar oxygen concentrations (P>.05). NA had a higher concentration of chlorine (P<.05), whereas LH had a lower amount of smear layer and a larger number of open dentin tubules than the other groups (P<.05). These were equivalent to each other (P>.05), except for HH, which also had a larger number of open dentin tubules than CG and NA (P<.05). PA 1% with a low concentration of hydrogen peroxide yielded a lower amount of smear layer and a larger number of open dentin tubules in the dentin of the post space when compared with PA 1% with a high concentration of hydrogen peroxide, despite maintaining a similar oxygen concentration in these dentin residues. Copyright © 2017 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

Quality Assurance of Rapid Diagnostic Tests for Malaria in Routine Patient Care in Rural Tanzania

PubMed Central

McMorrow, Meredith L.; Masanja, M. Irene; Kahigwa, Elizeus; Abdulla, Salim M. K.; Kachur, S. Patrick

2010-01-01

Histidine-rich protein II (HRP2)-based malaria rapid diagnostic tests (RDTs) have shown high sensitivity and specificity for detecting Plasmodium falciparum malaria in a variety of study settings. However, RDTs are susceptible to heat and humidity and variation in individual performance, which may affect their use in field settings. We evaluated sensitivity and specificity of RDTs during routine use for malaria case management in peripheral health facilities. From December 2007 to October 2008, HRP2-based ParaHIT-f RDTs were introduced in 12 facilities without available microscopy in Rufiji District, Tanzania. Health workers received a single day of instruction on how to perform an RDT and thick blood smear. Job aids, Integrated Management of Childhood Illness guidelines, and national malaria treatment algorithms were reviewed. For quality assurance (QA), thick blood smears for reference microscopy were collected for 2 to 3 days per week from patients receiving RDTs; microscopy was not routinely performed at the health facilities. Slides were stained and read centrally within 72 hours of collection by a reference microscopist. When RDT and blood smear results were discordant, blood smears were read by additional reference microscopists blinded to earlier results. Facilities were supervised monthly by the district laboratory supervisor or a member of the study team. Ten thousand six hundred fifty (10,650) patients were tested with RDTs, and 51.5% (5,488/10,650) had a positive test result. Blood smear results were available for 3,914 patients, of whom 40.1% (1,577/3,914) were positive for P. falciparum malaria. Overall RDT sensitivity was 90.7% (range by facility 85.7–96.5%) and specificity was 73.5% (range 50.0–84.3%). Sensitivity increased with increasing parasite density. Successful implementation of RDTs was achieved in peripheral health facilities with adequate training and supervision. Quality assurance is essential to the adequate performance of any laboratory test. Centralized staining and reading of blood smears provided useful monitoring of RDT performance. However, this level of QA may not be sustainable nationwide. PMID:20065013

Active case-finding for tuberculosis by mobile teams in Myanmar: yield and treatment outcomes.

PubMed

Myint, Ohnmar; Saw, Saw; Isaakidis, Petros; Khogali, Mohammed; Reid, Anthony; Hoa, Nguyen Binh; Kyaw, Thi Thi; Zaw, Ko Ko; Khaing, Tin Mi Mi; Aung, Si Thu

2017-06-02

Since 2005, the Myanmar National Tuberculosis Programme (NTP) has been implementing active case finding (ACF) activities involving mobile teams in hard-to-reach areas. This study revealed the contribution of mobile team activities to total tuberculosis (TB) case detection, characteristics of TB patients detected by mobile teams and their treatment outcomes. This was a descriptive study using routine programme data between October 2014 and December 2014. Mobile team activities were a one-stop service and included portable digital chest radiography (CXR) and microscopy of two sputum samples. The algorithm of the case detection included screening patients by symptoms, then by CXR followed by sputum microscopy for confirmation. Diagnosed patients were started on treatment and followed until a final outcome was ascertained. A total of 9 349 people with symptoms suggestive of TB were screened by CXR, with an uptake of 96.6%. Of those who were meant to undergo sputum smear microscopy, 51.4% had sputum examinations. Finally, 504 TB patients were identified by the mobile teams and the overall contribution to total TB case detection in the respective townships was 25.3%. Among total cases examined by microscopy, 6.4% were sputum smear positive TB. Treatment success rate was high as 91.8% in study townships compared to national rate 85% (2014 cohort). This study confirmed the feasibility and acceptability of ACF by mobile teams in hard-to-reach contexts, especially when equipped with portable, digital CXR machines that provided immediate results. However, the follow-up process of sputum examination created a significant barrier to confirmation of the diagnosis. In order to optimize the ACF through mobile team activity, future ACF activities were needed to be strengthened one stop service including molecular diagnostics or provision of sputum cups to all presumptive TB cases prior to CXR and testing if CXR suggestive of TB.

A comparative scanning electron microscopy evaluation of smear layer removal with apple vinegar and sodium hypochlorite associated with EDTA

PubMed Central

CANDEIRO, George Táccio de Miranda; de MATOS, Isabela Barbosa; da COSTA, Clarice Fernandes Eloy; FONTELES, Cristiane Sá Roriz; do VALE, Mônica Sampaio

2011-01-01

Objective The purpose of this study was to evaluate by scanning electron microscopy (SEM) the removal of smear layer from the middle and apical root thirds after use of different irrigating solutions. Material and Methods Forty roots of permanent human teeth had their canals instrumented and were randomly assigned to 4 groups (n=10), according to the irrigating solution: apple vinegar (group A), apple vinegar finished with 17% ethylenediaminetetraacetic acid (EDTA) (group B), 1% sodium hypochlorite (NaOCl) finished with 17% EDTA (group C) and saline (group D - control). After chemomechanical preparation, the roots were cleaved longitudinally and their middle and apical thirds were examined by SEM at ×1,000 magnification. Two calibrated examiners (kappa=0.92) analyzed the SEM micrographs qualitatively attributing scores that indicated the efficacy of the solutions in removing the smear layer from the surface of the dentin tubules (1 - poor, 2 - good and 3 - excellent). Data from the control and experimental groups were analyzed by the Kruskal-Wallis and Dunn's test, while the Wilcoxon test was used to compare the middle and apical thirds of the canals within the same group (α=0.05). Results The middle third presented less amount of smear layer than the apical third, regardless of the irrigant. There was statistically significant difference (p=0.0402) among the groups in the middle third. In the apical third, the apple vinegar/EDTA group showed the greatest removal of smear layer (p=0.0373). Conclusion Apple vinegar associated or not with EDTA was effective in removing smear layer when used as an endodontic irrigant. PMID:22231000

[Scanning electron microscopy investigation of canal cleaning after canal preparation with nickel titanium files].

PubMed

Brkanić, Tatjana; Ivana, Stojsin; Vukoje, Karolina; Zivković, Slavoljub

2010-01-01

Root canal preparation is the most important phase of endodontic procedure and it consists of adequate canal space cleaning and shaping. In recent years, rotary instruments and techniques have gained importance because of the great efficacy, speed and safety of the preparation procedure. The aim of this research was to investigate the influence of different NiTi files on the canal wall cleaning quality, residual dentine debris and smear layer. The research was conducted on extracted human teeth in vitro conditions. Teeth were divided in 7 main groups depending on the kind of instruments used for root canal preparation: ProTaper, GT, ProFile, K-3, FlexMaster, hand ProTaper and hand GT. Root canal preparation was accomplished by crown-down technique. Prepared samples were assessed on scanning electron microscopy JEOL, JSM-6460 LV. The evaluation of dentine debris was done with 500x magnification, and the evaluation of smear layer with 1,000 times magnification. Quantitive assessment of dentine debris and smear layer was done according to the criteria of Hulsmann. The least amount of debris and smear layer has been found in canals shaped with ProFile instruments, and the largest amount in canals shaped with FlexMaster instruments. Canal cleaning efficacy of hand GTand ProTaperfiles has been similar to cleaning efficacy of rotary NiTi files. Statistic analysis has shown a significant difference in amount of dentine debris and smear layer on the canal walls between sample groups. shaped with different instruments. Completely clean canals have not been found in any tested group of instruments. The largest amount of debris and smear layer has been found in the apical third of all canals. The design and the type of endodontic instruments influence the efficacy of the canal cleaning.

Dual function of EDTA with silver nanoparticles for root canal treatment-A novel modification.

PubMed

Martinez-Andrade, Juan M; Avalos-Borja, Miguel; Vilchis-Nestor, Alfredo R; Sanchez-Vargas, Luis O; Castro-Longoria, Ernestina

2018-01-01

The chelating and antimicrobial capacity of a novel modification of 17% EDTA with silver nanoparticles (AgNPs) (EDTA-AgNPs) was evaluated in-vitro for root canal treatment (RCT). The EDTA-AgNPs solution was characterized by UV-Vis spectroscopy, ζ-potential and high-resolution transmission electron microscopy (HRTEM). Antimicrobial capacity was evaluated against Candida albicans and Staphylococcus aureus in planktonic and biofilm cells by broth macrodilution (24 h) and XTT assays, (1, 10 and 30 min) respectively. The chelating capacity of EDTA-AgNPs was assessed indirectly (smear layer removal) and directly (demineralizing effect) in bovine dentin at two silver concentrations, 16 and 512 μg/ml at 1 and 10 minutes of exposure time. Smear layer removal was evaluated by atomic force microscopy (AFM) and scanning electron microscopy (SEM). The demineralizing effect was determined by atomic absorption spectroscopy (AAS), microhardness test (MH) and X-ray diffractometer (XRD). Synthesized AgNPs were quasi-spherical in shape with an average size of 13.09 ± 8.05 nm. 17% EDTA-AgNPs was effective to inhibit C. albicans and S. aureus in planktonic and biofilm cultures. The smear layer removal and demineralizing effect were similar between 17% EDTA-AgNPs and 17% EDTA treatments. The 17% EDTA-AgNPs solution proved to be an effective antimicrobial agent, and has a similar chelating capacity to 17% EDTA alone. These in-vitro studies strongly suggest that EDTA-AgNPs could be used for effective smear layer removal, having an antimicrobial effect at the same time during RCT.

Impact of Laboratory Practice Changes on the Diagnosis of Tuberculosis with the Introduction of Xpert MTB/RIF in Kiribati.

PubMed

Tonganibeia, Alfred; Harries, Anthony D; Merilles, Onofre Edwin A; Tarataake, Tekaibeti; Tiira, Teatao; Kienene, Takeieta

2018-02-01

The Republic of Kiribati, Central Pacific, has the largest tuberculosis epidemic in the region. There is a national tuberculosis control program, which has used smear microscopy for acid-fast bacilli as the main diagnostic tool for many years. In 2015, an Xpert MTB/RIF machine was procured and became functional within the tuberculosis hospital. The aim of this cross-sectional study, using routinely collected data, was to determine the effects of introducing Xpert MTB/RIF on laboratory smear microscopy practices and the pattern of registered tuberculosis cases. Between February 2015 and January 2016, there were 220 Xpert MTB/RIF assays performed with 6.4% errors and 15% detection of Mycobacterium tuberculosis: one patient showed rifampicin-resistance. One year before and after introducing Xpert MTB/RIF, the number of presumptive tuberculosis patients increased by 9% from 2,138 to 2,322. There were no changes in demographic characteristics, smear-positive results, or acid-fast bacilli grade between the two periods. The number of specimens cultured for Mycobacterium tuberculosis significantly declined from 638 to zero, with 76 positive MTB cultures before and none after introducing Xpert MTB/RIF. There was a significant change in the profile of registered tuberculosis cases with more children (34% versus 21%) and fewer bacteriologically-confirmed cases (29% versus 43%) - P < .001. Since the deployment of Xpert MTB/RIF in Kiribati, there have been a small number of assays performed and this has been associated with no adverse effects on smear microscopy, a stoppage in mycobacterial cultures, and a change in the types and categories of diagnosed tuberculosis.

Dual function of EDTA with silver nanoparticles for root canal treatment–A novel modification

PubMed Central

Martinez-Andrade, Juan M.; Avalos-Borja, Miguel; Vilchis-Nestor, Alfredo R.; Sanchez-Vargas, Luis O.

2018-01-01

The chelating and antimicrobial capacity of a novel modification of 17% EDTA with silver nanoparticles (AgNPs) (EDTA-AgNPs) was evaluated in-vitro for root canal treatment (RCT). The EDTA-AgNPs solution was characterized by UV-Vis spectroscopy, ζ-potential and high-resolution transmission electron microscopy (HRTEM). Antimicrobial capacity was evaluated against Candida albicans and Staphylococcus aureus in planktonic and biofilm cells by broth macrodilution (24 h) and XTT assays, (1, 10 and 30 min) respectively. The chelating capacity of EDTA-AgNPs was assessed indirectly (smear layer removal) and directly (demineralizing effect) in bovine dentin at two silver concentrations, 16 and 512 μg/ml at 1 and 10 minutes of exposure time. Smear layer removal was evaluated by atomic force microscopy (AFM) and scanning electron microscopy (SEM). The demineralizing effect was determined by atomic absorption spectroscopy (AAS), microhardness test (MH) and X-ray diffractometer (XRD). Synthesized AgNPs were quasi-spherical in shape with an average size of 13.09 ± 8.05 nm. 17% EDTA-AgNPs was effective to inhibit C. albicans and S. aureus in planktonic and biofilm cultures. The smear layer removal and demineralizing effect were similar between 17% EDTA-AgNPs and 17% EDTA treatments. The 17% EDTA-AgNPs solution proved to be an effective antimicrobial agent, and has a similar chelating capacity to 17% EDTA alone. These in-vitro studies strongly suggest that EDTA-AgNPs could be used for effective smear layer removal, having an antimicrobial effect at the same time during RCT. PMID:29346398

Use of Transrenal DNA for the Diagnosis of Extrapulmonary Tuberculosis in Children: a Case of Tubercular Otitis Media

PubMed Central

Petrucci, Roberta; Corsini, Ilaria; Visciotti, Francesca; Pirodda, Antonio; Cazzato, Salvatore; Landini, Maria Paola; Dal Monte, Paola

2014-01-01

The diagnosis of tuberculosis (TB) is difficult in children, especially for smear-negative pulmonary and extrapulmonary TB, which are common at this age. We report an 11-year-old girl with TB otitis media with negative smear microscopy and Xpert MTB/RIF but positive Mycobacterium tuberculosis-specific transrenal DNA (Tr-MTB-DNA) test results and culture for M. tuberculosis. PMID:25339389

Fragmentation of Red Blood Cells Caused Pseudothrombocytosis in a Patient.

PubMed

Tang, Wenjun; Tang, Chunhua; Zheng, Jianfeng; He, Yongling; Lu, Fangfang

2018-06-01

Pseudothrombocytopenia, caused by platelet (PLT) clumping, is often found in clinical studies [1]. However, pseudothrombocytosis resulting from the fragmentation of red blood cells (RBCs) is a very rare phenomenon. EDTA-K2 anticoagulation was used on a sample of venous blood extracted from the patient. A Symex XN9000 automatic blood analyzer was used to conduct CBC + DIFF mode and CBC + DIFF + RET mode tests, stained smear microscopy. The Symex XN9000 automatic blood analyzer was used to conduct CBC + DIFF mode test; PLTs were measured at 570 x 109/L. Stained smear microscopy revealed the number of PLTs did not conform to the instrument measured 570 x 109/L. "RET" alarm instrument, switch to CBC + DIFF + RET mode for testing. The second test result showed PLTs at 128 x 109/L, which accords with artificial microscopy. This was a case of a very rare phenomenon: the fragmentation of RBCs caused pseudothrombocytosis.

Dynamic imaging model and parameter optimization for a star tracker.

PubMed

Yan, Jinyun; Jiang, Jie; Zhang, Guangjun

2016-03-21

Under dynamic conditions, star spots move across the image plane of a star tracker and form a smeared star image. This smearing effect increases errors in star position estimation and degrades attitude accuracy. First, an analytical energy distribution model of a smeared star spot is established based on a line segment spread function because the dynamic imaging process of a star tracker is equivalent to the static imaging process of linear light sources. The proposed model, which has a clear physical meaning, explicitly reflects the key parameters of the imaging process, including incident flux, exposure time, velocity of a star spot in an image plane, and Gaussian radius. Furthermore, an analytical expression of the centroiding error of the smeared star spot is derived using the proposed model. An accurate and comprehensive evaluation of centroiding accuracy is obtained based on the expression. Moreover, analytical solutions of the optimal parameters are derived to achieve the best performance in centroid estimation. Finally, we perform numerical simulations and a night sky experiment to validate the correctness of the dynamic imaging model, the centroiding error expression, and the optimal parameters.

A comparative study of blood smear, QBC and antigen detection for diagnosis of malaria.

PubMed

Parija, S C; Dhodapkar, Rahul; Elangovan, Subashini; Chaya, D R

2009-01-01

Rapid diagnosis is prerequisite for effective treatment and reducing mortality and morbidity of malaria. This study was taken up to compare the efficacy of various methods available, i.e., thick and thin smear, quantitative buffy coat (QBC), plasmodium lactate dehydrogenase and aldolase in blood of patient. A total of 411 samples were collected from patients presenting with classic symptoms of malaria. For traditional microscopy; thick and thin smears were prepared and stained with Leishman's stain, taking thick smear as gold standard, thin smear had a sensitivity and specificity of 54.8% and 100%, respectively. QBC and antigen detection was done using commercially available kits; out of 411 samples, QBC and Malariagen were positive in 66 and 62 cases, with a sensitivity of 78% and 75%, respectively. Leishman's thick smear, although cost effective, is difficult to interpret for inexperienced microscopists; so if facilities are available, QBC should be used for routine diagnosis. In places where facilities are not available, rapid, simple and easy to interpret antigen detection test can be used despite low sensitivity.

Simplified Pan-species Real-time PCR-based Detection of Plasmodium Spp. in Blood Smear

PubMed Central

HASSANPOUR, Gholamreza; MIRHENDI, Hossein; MOHEBALI, Mehdi; RAEISI, Ahmad; ZERAATI, Hojjat; KESHAVARZ, Hossein

2016-01-01

Background: We aimed to quicken and simplify the detection of Plasmodium in blood samples by developing and testing a pan-Plasmodium real-time PCR for accurate screening of individuals suspected of malaria. Methods: A single primer/probe set for pan-species Plasmodium-specific real time PCR targeting a conserved region of the small subunit 18S ribosomal DNA was designed and evaluated for rapid diagnosis and screening of malaria infections using dried blood smears. FTA cards were used for rapid and simple DNA extraction. Results: The primers and probes showed a positive response with the DNA extracted from bloods infected with P. falciparum and P. vivax but not with DNA extracted from various smears from uninfected blood samples. Seven positive cases positive by both microscopy and nested PCR were found among 280 blood samples taken from in South and Southeast Iran. Five samples were identified as positive for P. vivax and two as positive for P. falciparum. All positive samples were positive by real-time PCR. Furthermore, all 38-blood samples positive by microscopy were positive by real-time PCR. No microscopy-negative samples were positive by real-time PCR. Conclusion: By using a simple FTA card for DNA extraction and by application of the real-time PCR developed in this study, sensitivity similar to nested-PCR and microscopy was achieved. This format simplifies the detection of Plasmodium in large numbers of samples. PMID:28127357

Evaluation of mobile digital light-emitting diode fluorescence microscopy in Hanoi, Viet Nam.

PubMed

Chaisson, L H; Reber, C; Phan, H; Switz, N; Nilsson, L M; Myers, F; Nhung, N V; Luu, L; Pham, T; Vu, C; Nguyen, H; Nguyen, A; Dinh, T; Nahid, P; Fletcher, D A; Cattamanchi, A

2015-09-01

Hanoi Lung Hospital, Hanoi, Viet Nam. To compare the accuracy of CellScopeTB, a manually operated mobile digital fluorescence microscope, with conventional microscopy techniques. Patients referred for sputum smear microscopy to the Hanoi Lung Hospital from May to September 2013 were included. Ziehl-Neelsen (ZN) smear microscopy, conventional light-emitting diode (LED) fluorescence microscopy (FM), CellScopeTB-based LED FM and Xpert(®) MTB/RIF were performed on sputum samples. The sensitivity and specificity of microscopy techniques were determined in reference to Xpert results, and differences were compared using McNemar's paired test of proportions. Of 326 patients enrolled, 93 (28.5%) were Xpert-positive for TB. The sensitivity of ZN microscopy, conventional LED FM, and CellScopeTB-based LED FM was respectively 37.6% (95%CI 27.8-48.3), 41.9% (95%CI 31.8-52.6), and 35.5% (95%CI 25.8-46.1). The sensitivity of CellScopeTB was similar to that of conventional LED FM (difference -6.5%, 95%CI -18.2 to 5.3, P = 0.33) and ZN microscopy (difference -2.2%, 95%CI -9.2 to 4.9, P = 0.73). The specificity was >99% for all three techniques. CellScopeTB performed similarly to conventional microscopy techniques in the hands of experienced TB microscopists. However, the sensitivity of all sputum microscopy techniques was low. Options enabled by digital microscopy, such as automated imaging with real-time computerized analysis, should be explored to increase sensitivity.

Smear plus Detect-TB for a sensitive diagnosis of pulmonary tuberculosis: a cost-effectiveness analysis in an incarcerated population.

PubMed

Schmid, Karen Barros; Scherer, Luciene; Barcellos, Regina Bones; Kuhleis, Daniele; Prestes, Isaías Valente; Steffen, Ricardo Ewbank; Dalla Costa, Elis Regina; Rossetti, Maria Lucia Rosa

2014-12-16

Prison conditions can favor the spread of tuberculosis (TB). This study aimed to evaluate in a Brazilian prison: the performance and accuracy of smear, culture and Detect-TB; performance of smear plus culture and smear plus Detect-TB, according to different TB prevalence rates; and the cost-effectiveness of these procedures for pulmonary tuberculosis (PTB) diagnosis. This paper describes a cost-effectiveness study. A decision analytic model was developed to estimate the costs and cost-effectiveness of five routine diagnostic procedures for diagnosis of PTB using sputum specimens: a) Smear alone, b) Culture alone, c) Detect-TB alone, d) Smear plus culture and e) Smear plus Detect-TB. The cost-effectiveness ratio of costs were evaluated per correctly diagnosed TB case and all procedures costs were attributed based on the procedure costs adopted by the Brazilian Public Health System. A total of 294 spontaneous sputum specimens from patients suspected of having TB were analyzed. The sensibility and specificity were calculated to be 47% and 100% for smear; 93% and 100%, for culture; 74% and 95%, for Detect-TB; 96% and 100%, for smear plus culture; and 86% and 95%, for smear plus Detect-TB. The negative and positive predictive values for smear plus Detect-TB, according to different TB prevalence rates, ranged from 83 to 99% and 48 to 96%, respectively. In a cost-effectiveness analysis, smear was both less costly and less effective than the other strategies. Culture and smear plus culture were more effective but more costly than the other strategies. Smear plus Detect-TB was the most cost-effective method. The Detect-TB evinced to be sensitive and effective for the PTB diagnosis when applied with smear microscopy. Diagnostic methods should be improved to increase TB case detection. To support rational decisions about the implementation of such techniques, cost-effectiveness studies are essential, including in prisons, which are known for health care assessment problems.

Papanicolaou stain: Is it economical to switch to rapid, economical, acetic acid, papanicolaou stain?

PubMed

Dighe, Swati B; Ajit, Dulhan; Pathuthara, Saleem; Chinoy, Roshni

2006-01-01

To standardize an inexpensive and rapid Papanicolaou staining technique with limited ethanol usage. Smears from 200 patients were collected (2 per patient) and fixed in methanol. Half were subjected to conventional Papanicolaou and half to stain ing with rapid, economical, acetic acid Papanicolaou (REAP) stain. In REAP, pre-OG6 and post-OG6 and post-EA36 ethanol baths were replaced by 1% acetic acid and Scott's tap water with tap water. Hematoxylin was preheated to 60 degrees C. Final dehydration was with methanol. REAP smears were compared with Papanicolaou smears for optimal cytoplasmic and nuclear staining, stain preservation, cost and turnaround time. With the REAP method, cytoplasmic and nuclear staining was optimal in 181 and 192 cases, respectively. The staining time was considerably reduced, to 3 minutes, and the cost per smear was reduced to one fourth. The staining quality remained good in all the smears for > 2 years. REAP is a rapid, cost-effective alternative to Papanicolaou stain. Though low stain penetration in large cell clusters is a limitation, final interpretation was not compromised.

Impact of Xpert MTB/RIF rollout on management of tuberculosis in a South African community.

PubMed

Schmidt, B-M; Geldenhuys, H; Tameris, M; Luabeya, A; Mulenga, H; Bunyasi, E; Scriba, T; Hatherill, M

2017-11-27

The Xpert MTB/RIF test shortens the time to microbiological confirmation of pulmonary tuberculosis (TB) under research conditions. To evaluate the field impact of Xpert MTB/RIF rollout on TB diagnostic yield and time to treatment in a South African (SA) community. We compared TB investigation outcomes for 6-month calendar periods before and after Xpert MTB/RIF rollout in a semi-rural area of SA. The proportion of adult patients who tested positive by sputum smear microscopy, liquid culture or Xpert MTB/RIF and the proportion of positive sputum smear, liquid culture or Xpert MTB/RIF tests were compared. Secondary outcomes included time to laboratory diagnosis and treatment initiation. Data were collected from the National Health Laboratory Service database and from the Western Cape Provincial Department of Health TB register. Regional rollout of Xpert MTB/RIF testing occurred in 2013. Of the 15 629 patients investigated in the post-rollout period, 7.9% tested positive on GeneXpert, compared with 6.4% of the 10 741 investigated in the pre-rollout period who tested positive by sputum smear microscopy (p<0.001). Median laboratory processing time was <1 day for Xpert MTB/RIF (interquartile range (IQR) 0 - 1) compared with 1 day (IQR 0 - 16) for sputum smear microscopy (p=0.001). The median time to TB treatment initiation was 4 days (IQR 2 - 8) after rollout compared with 5 days (IQR 2 - 14) before (p=0.001). Patients investigated for suspected pulmonary TB were more likely to be diagnosed after rollout of Xpert MTB/RIF testing, although the benefit to diagnostic yield was modest, and Xpert MTB/RIF testing was associated with a marginal improvement in time to treatment initiation.

Challenges in Routine Implementation and Quality Control of Rapid Diagnostic Tests for Malaria–Rufiji District, Tanzania

PubMed Central

McMorrow, Meredith L.; Masanja, M. Irene; Abdulla, Salim M. K.; Kahigwa, Elizeus; Kachur, S. Patrick

2018-01-01

Rapid diagnostic tests (RDTs) represent an alternative to microscopy for malaria diagnosis and have shown high sensitivity and specificity in a variety of study settings. Current World Health Organization (WHO) guidelines for quality control of RDTs provide detailed instructions on pre-field testing, but offer little guidance for quality assurance once RDTs are deployed in health facilities. From September 2006 to April 2007, we introduced a histidine-rich protein II (HRP2)-based RDT (Paracheck) for suspected malaria cases five years of age and older in nine health facilities in Rufiji District, Tanzania, to assess sensitivity and specificity of RDTs in routine use at rural health facilities. Thick blood smears were collected for all patients tested with RDTs and stained and read by laboratory personnel in each facility. Thick smears were subsequently reviewed by a reference microscopist to determine RDT sensitivity and specificity. In all nine health facilities, there were significant problems with the quality of staining and microscopy. Sensitivity and specificity of RDTs were difficult to assess given the poor quality of routine blood smear staining. Mean operational sensitivity of RDTs based on reference microscopy was 64.8%, but varied greatly by health facility, range 18.8–85.9%. Sensitivity of RDTs increased with increasing parasite density. Specificity remained high at 87.8% despite relatively poor slide quality. Institution of quality control of RDTs based on poor quality blood smear staining may impede reliable measurement of sensitivity and specificity and undermine confidence in the new diagnostic. There is an urgent need for the development of alternative quality control procedures for rapid diagnostic tests that can be performed at the facility level. PMID:18784230

Conformational Smear Characterization and Binning of Single-Molecule Conductance Measurements for Enhanced Molecular Recognition.

PubMed

Korshoj, Lee E; Afsari, Sepideh; Chatterjee, Anushree; Nagpal, Prashant

2017-11-01

Electronic conduction or charge transport through single molecules depends primarily on molecular structure and anchoring groups and forms the basis for a wide range of studies from molecular electronics to DNA sequencing. Several high-throughput nanoelectronic methods such as mechanical break junctions, nanopores, conductive atomic force microscopy, scanning tunneling break junctions, and static nanoscale electrodes are often used for measuring single-molecule conductance. In these measurements, "smearing" due to conformational changes and other entropic factors leads to large variances in the observed molecular conductance, especially in individual measurements. Here, we show a method for characterizing smear in single-molecule conductance measurements and demonstrate how binning measurements according to smear can significantly enhance the use of individual conductance measurements for molecular recognition. Using quantum point contact measurements on single nucleotides within DNA macromolecules, we demonstrate that the distance over which molecular junctions are maintained is a measure of smear, and the resulting variance in unbiased single measurements depends on this smear parameter. Our ability to identify individual DNA nucleotides at 20× coverage increases from 81.3% accuracy without smear analysis to 93.9% with smear characterization and binning (SCRIB). Furthermore, merely 7 conductance measurements (7× coverage) are needed to achieve 97.8% accuracy for DNA nucleotide recognition when only low molecular smear measurements are used, which represents a significant improvement over contemporary sequencing methods. These results have important implications in a broad range of molecular electronics applications from designing robust molecular switches to nanoelectronic DNA sequencing.

Direct detection of Mycobacterium tuberculosis rifampin resistance in bio-safe stained sputum smears.

PubMed

Lavania, Surabhi; Anthwal, Divya; Bhalla, Manpreet; Singh, Nagendra; Haldar, Sagarika; Tyagi, Jaya Sivaswami

2017-01-01

Direct smear microscopy of sputum forms the mainstay of TB diagnosis in resource-limited settings. Stained sputum smear slides can serve as a ready-made resource to transport sputum for molecular drug susceptibility testing. However, bio-safety is a major concern during transport of sputum/stained slides and for laboratory workers engaged in processing Mycobacterium tuberculosis infected sputum specimens. In this study, a bio-safe USP (Universal Sample Processing) concentration-based sputum processing method (Bio-safe method) was assessed on 87 M. tuberculosis culture positive sputum samples. Samples were processed for Ziehl-Neelsen (ZN) smear, liquid culture and DNA isolation. DNA isolated directly from sputum was subjected to an IS6110 PCR assay. Both sputum DNA and DNA extracted from bio-safe ZN concentrated smear slides were subjected to rpoB PCR and simultaneously assessed by DNA sequencing for determining rifampin (RIF) resistance. All sputum samples were rendered sterile by Bio-safe method. Bio-safe smears exhibited a 5% increment in positivity over direct smear with a 14% increment in smear grade status. All samples were positive for IS6110 and rpoB PCR. Thirty four percent samples were RIF resistant by rpoB PCR product sequencing. A 100% concordance (κ value = 1) was obtained between sequencing results derived from bio-safe smear slides and bio-safe sputum. This study demonstrates that Bio-safe method can address safety issues associated with sputum processing, provide an efficient alternative to sample transport in the form of bio-safe stained concentrated smear slides and can also provide information on drug (RIF) resistance by direct DNA sequencing.

Time to sputum conversion in smear positive pulmonary TB patients on category I DOTS and factors delaying it.

PubMed

Parikh, Raunak; Nataraj, Gita; Kanade, Swapna; Khatri, Vijay; Mehta, Preeti

2012-08-01

Sputum smear positive pulmonary tuberculosis patients expel infectious viable bacilli for a period following commencement of treatment. Patients on Directly Observed Treatment Shortcourse (DOTS) receive intermittent therapy with multidrug regimen. To determine the time to sputum smear and culture conversion following initiation of DOTS treatment and study the factors that influence it. A prospective study was undertaken at a tertiary care teaching hospital in Mumbai over a one year period on a cohort of 71 sputum smear positive patients on Category I DOTS treatment. Patients were followed up weekly for upto 20 weeks or until they underwent smear and culture conversion whichever was earlier. At each follow up, specimens were collected and processed for microscopy and culture using standard protocol. 60/71 [84.55%] patients completed the study. 56/60 [93.3%] patients underwent sputum smear and culture conversion. The median time to smear and culture conversion was end of 5th week [day 35] and 6 1/2 weeks [day 45] respectively. Univariate and stepwise regression analysis showed that patients who had cavitatory disease, high pretreatment smear grade and a past history of tuberculosis were more likely to undergo delayed or nonconversion [P < 0.05]. The time to sputum smear and culture conversion under DOTS is similar to previous antituberculosis regimens. Since viable bacilli continue to be expelled for upto two months, infection control measures should be maintained for such time. Patients with cavitatory disease, high pretreatment smear grade or a past history of tuberculosis need to be monitored more closely.

Direct detection of Mycobacterium tuberculosis rifampin resistance in bio-safe stained sputum smears

PubMed Central

Lavania, Surabhi; Anthwal, Divya; Bhalla, Manpreet; Singh, Nagendra; Haldar, Sagarika; Tyagi, Jaya Sivaswami

2017-01-01

Direct smear microscopy of sputum forms the mainstay of TB diagnosis in resource-limited settings. Stained sputum smear slides can serve as a ready-made resource to transport sputum for molecular drug susceptibility testing. However, bio-safety is a major concern during transport of sputum/stained slides and for laboratory workers engaged in processing Mycobacterium tuberculosis infected sputum specimens. In this study, a bio-safe USP (Universal Sample Processing) concentration-based sputum processing method (Bio-safe method) was assessed on 87 M. tuberculosis culture positive sputum samples. Samples were processed for Ziehl-Neelsen (ZN) smear, liquid culture and DNA isolation. DNA isolated directly from sputum was subjected to an IS6110 PCR assay. Both sputum DNA and DNA extracted from bio-safe ZN concentrated smear slides were subjected to rpoB PCR and simultaneously assessed by DNA sequencing for determining rifampin (RIF) resistance. All sputum samples were rendered sterile by Bio-safe method. Bio-safe smears exhibited a 5% increment in positivity over direct smear with a 14% increment in smear grade status. All samples were positive for IS6110 and rpoB PCR. Thirty four percent samples were RIF resistant by rpoB PCR product sequencing. A 100% concordance (κ value = 1) was obtained between sequencing results derived from bio-safe smear slides and bio-safe sputum. This study demonstrates that Bio-safe method can address safety issues associated with sputum processing, provide an efficient alternative to sample transport in the form of bio-safe stained concentrated smear slides and can also provide information on drug (RIF) resistance by direct DNA sequencing. PMID:29216262

Tip vortices in the actuator line model

NASA Astrophysics Data System (ADS)

Martinez, Luis; Meneveau, Charles

2017-11-01

The actuator line model (ALM) is a widely used tool to represent the wind turbine blades in computational fluid dynamics without the need to resolve the full geometry of the blades. The ALM can be optimized to represent the `correct' aerodynamics of the blades by choosing an appropriate smearing length scale ɛ. This appropriate length scale creates a tip vortex which induces a downwash near the tip of the blade. A theoretical frame-work is used to establish a solution to the induced velocity created by a tip vortex as a function of the smearing length scale ɛ. A correction is presented which allows the use of a non-optimal smearing length scale but still provides the downwash which would be induced using the optimal length scale. Thanks to the National Science Foundation (NSF) who provided financial support for this research via Grants IGERT 0801471, IIA-1243482 (the WINDINSPIRE project) and ECCS-1230788.

Assessment of the sensitivity and specificity of Xpert MTB/RIF assay as an early sputum biomarker of response to tuberculosis treatment.

PubMed

Friedrich, Sven O; Rachow, Andrea; Saathoff, Elmar; Singh, Kasha; Mangu, Chacha D; Dawson, Rodney; Phillips, Patrick Pj; Venter, Amour; Bateson, Anna; Boehme, Catharina C; Heinrich, Norbert; Hunt, Robert D; Boeree, Martin J; Zumla, Alimuddin; McHugh, Timothy D; Gillespie, Stephen H; Diacon, Andreas H; Hoelscher, Michael

2013-08-01

An accurate biomarker is urgently needed to monitor the response to treatment in patients with pulmonary tuberculosis. The Xpert MTB/RIF assay is a commercially available real-time PCR that can be used to detect Mycobacterium-tuberculosis-specific DNA sequences in sputum samples. We therefore evaluated this assay with serial sputum samples obtained over 26 weeks from patients undergoing treatment for tuberculosis. We analysed sputum samples from 221 patients with smear-positive tuberculosis enrolled at two sites (Cape Town, South Africa, and Mbeya, Tanzania) of a multicentre randomised clinical trial REMoxTB of antituberculosis treatment on a weekly basis (weeks 0 to 8), then at weeks 12, 17, 22, and 26 after treatment initiation. The Xpert MTB/RIF results over time were compared with the results of standard smear microscopy and culture methods. We obtained and analysed 2741 sputum samples from 221 patients. The reduction in positivity rates with Xpert MTB/RIF were slower than those with the standard methods. At week 8, positive results were obtained for 62 (29%) of 212 sputum samples with smear microscopy, 46 (26%) of 175 with solid culture (Löwenstein-Jensen medium), 77 (42%) of 183 with liquid culture (Bactec MGIT960 system), and 174 (84%) of 207 with Xpert MTB/RIF; at 26 weeks, positive results were obtained for ten (5%) of 199, four (3%) of 157, seven (4%) of 169, and 22 (27%) of 83 sputum samples, respectively. The reduction in detection of quantitative M tuberculosis DNA with Xpert MTB/RIF correlated with smear grades (ρ=-0·74; p<0·0001), solid culture grades (ρ=-0·73; p<0·0001), and time to liquid culture positivity (ρ=0·73; p<0·0001). Compared with the combined binary smear and culture results as a reference standard, the Xpert MTB/RIF assay had high sensitivity (97·0%, 95% CI 95·8-97·9), but poor specificity (48·6%, 45·0-52·2). The poor specificity precludes the use of the Xpert MTB/RIF assay as a biomarker for monitoring tuberculosis treatment, and should not replace standard smear microscopy and culture. Global Alliance for TB Drug Development, Bill & Melinda Gates Foundation, UK Medical Research Council, German Ministry of Science and Technology. Copyright © 2013 Elsevier Ltd. All rights reserved.

Use of transrenal DNA for the diagnosis of extrapulmonary tuberculosis in children: a case of tubercular otitis media.

PubMed

Petrucci, Roberta; Lombardi, Giulia; Corsini, Ilaria; Visciotti, Francesca; Pirodda, Antonio; Cazzato, Salvatore; Landini, Maria Paola; Dal Monte, Paola

2015-01-01

The diagnosis of tuberculosis (TB) is difficult in children, especially for smear-negative pulmonary and extrapulmonary TB, which are common at this age. We report an 11-year-old girl with TB otitis media with negative smear microscopy and Xpert MTB/RIF but positive Mycobacterium tuberculosis-specific transrenal DNA (Tr-MTB-DNA) test results and culture for M. tuberculosis. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

The method of segmentation of leukocytes in information-measuring systems on the basis of light microscopy

NASA Astrophysics Data System (ADS)

Nikitaev, V. G.; Pronichev, A. N.; Polyakov, E. V.; Zaharenko, Yu V.

2018-01-01

The paper considers the problem of leukocytes segmentation in microscopic images of bone marrow smears for automated diagnosis of the blood system diseases. The method was proposed to solve the problem of segmentation of contacting leukocytes in images of bone marrow smears. The method is based on the analysis of structure of objects of a separation and distances filter in combination with the watershed method and distance transformation method.

Inpatient mortality in children with clinically diagnosed malaria as compared with microscopically confirmed malaria.

PubMed

Opoka, Robert O; Xia, Zongqi; Bangirana, Paul; John, Chandy C

2008-04-01

Inpatient treatment for malaria without microscopic confirmation of the diagnosis occurs commonly in sub-Saharan Africa. Differences in mortality in children who are tested by microscopy for Plasmodium falciparum infection as compared with those not tested are not well characterized. A retrospective chart review was conducted of all children up to 15 years of age admitted to Mulago Hospital, Kampala, Uganda from January 2002 to July 2005, with a diagnosis of malaria and analyzed according to microscopy testing for P. falciparum. A total of 23,342 children were treated for malaria during the study period, 991 (4.2%) of whom died. Severe malarial anemia in 7827 (33.5%) and cerebral malaria in 1912 (8.2%) were the 2 common causes of malaria-related admissions. Children who did not receive microscopy testing had a higher case fatality rate than those with a positive blood smear (7.5% versus 3.2%, P < 0.001). After adjustment for age, malaria complications, and comorbid conditions, children who did not have microscopy performed or had a negative blood smear had a higher risk of death than those with a positive blood smear [odds ratio (OR): 3.49, 95% confidence interval (CI): 2.88-4.22, P < 0.001; and OR: 1.59, 95% CI: 1.29-1.96, P < 0.001, respectively]. Diagnosis of malaria in the absence of microscopic confirmation is associated with significantly increased mortality in hospitalized Ugandan children. Inpatient diagnosis of malaria should be supported by microscopic or rapid diagnostic test confirmation.

Microscopic diagnosis of dysbacteriosis in stained vaginal smears in clinical practice.

PubMed

Verbruggen, Banut-Sabine M; Boon, Mathilde E; Melkerl, Peter; van Haaften, Maarten; Heintz, A Peter M

2006-10-01

Dysbacteriosis is a microscopical diagnosis. In women with dysbacteriosis, an overgrowth of coccoid bacteria and almost a complete absence of lactobacilli are observed in the (stained) vaginal smear. The aim of this study was to determine the accuracy of this microscopic diagnosis in clinical practice. The analysis concerned 342 consecutive cases in which the microscopy of the stained smears was performed by general practitioners trained in diagnosing dysbacteriosis. These smears were sent to the pathologist for confirmation of the microscopical diagnosis of the clinician. The cytological diagnoses of the pathologist, sometimes performed on restained slides when the quality of the staining was substandard, were considered as the "gold standard." In 92 of the 342 cases, dysbacteriosis was unequivocally established by the pathologist. Sensitivity and specificity of the microscopical diagnoses of the clinicians were 40% and 85%, respectively. There were 37 false-positive and 54 false-negative diagnoses of dysbacteriosis rendered by the clinicians. The most frequent reason for a false-negative diagnosis was an excess of lactobacilli in the smear. This study shows that even in stained smears it is difficult for clinicians to render a correct evaluation of the status of the vaginal flora.

Bronchoscopy for the diagnosis of pulmonary tuberculosis in patients with negative sputum smear microscopy results.

PubMed

Jacomelli, Márcia; Silva, Priscila Regina Alves Araújo; Rodrigues, Ascedio Jose; Demarzo, Sergio Eduardo; Seicento, Márcia; Figueiredo, Viviane Rossi

2012-01-01

To evaluate the diagnostic accuracy of bronchoscopy in patients with clinical or radiological suspicion of tuberculosis who were unable to produce sputum or with negative sputum smear microscopy results. A prospective cross-sectional study involving 286 patients under clinical or radiological suspicion of having pulmonary tuberculosis and submitted to bronchoscopy-BAL and transbronchial biopsy (TBB). The BAL specimens were submitted to direct testing and culture for AFB and fungi, whereas the TBB specimens were submitted to histopathological examination. Of the 286 patients studied, 225 (79%) were diagnosed on the basis of bronchoscopic findings, as follows: pulmonary tuberculosis, in 127 (44%); nonspecific chronic inflammation, in 51 (18%); pneumocystis, fungal infections, or nocardiosis, in 20 (7%); bronchiolitis obliterans organizing pneumonia, alveolites, or pneumoconiosis, in 14 (5%); lung or metastatic neoplasms, in 7 (2%); and nontuberculous mycobacterium infections, in 6 (2%). For the diagnosis of tuberculosis, BAL showed a sensitivity and a specificity of 60% and 100%, respectively. Adding the TBB findings significantly increased this sensitivity (to 84%), as did adding the post-bronchoscopy sputum smear microscopy results (total sensitivity, 94%). Minor post-procedure complications occurred in 5.6% of the cases. Bronchoscopy is a reliable method for the diagnosis of pulmonary tuberculosis, with low complication rates. The combination of TBB and BAL increases the sensitivity of the method and facilitates the differential diagnosis with other diseases.

The remote diagnosis of malaria using telemedicine or e-mailed images.

PubMed

Murray, Clinton K; Mody, Rupal M; Dooley, David P; Hospenthal, Duane R; Horvath, Lynn L; Moran, Kimberly A; Muntz, Ronald W

2006-12-01

We determined the ability of blinded remote expert microscopy to identify malaria parasites through transmission of malaria smear images via telemedicine and as e-mail attachments. Protocols for malaria smear transmission included: (1) transmission of sender-selected televised smears at various bandwidths (Bw), (2) transmission of remote reader-directed televised smears at various Bw, and (3) transmission of digital photomicrographs as e-mail attachments. Twenty (14%) of 147 sender-selected, and 13 (6%) of 221 reader-directed, images were deemed unreadable by slide readers. The presence or absence of malaria was correctly identified in 98% of the remaining images. Sixty-four (34%) of 190 digital microphotographs were deemed unreadable, while the presence or absence of malaria was correctly identified in 100% of the remaining images. Correct speciation ranged from 45% to 83% across various transmission methods and Bw. The use of telemedicine and e-mail technology shows promise for the remote diagnosis of malaria.

Evaluation of Xpert MTB/RIF assay in children with presumed pulmonary tuberculosis in Papua New Guinea.

PubMed

Kasa Tom, Sharon; Welch, Henry; Kilalang, Cornelia; Tefuarani, Nakapi; Vince, John; Lavu, Evelyn; Johnson, Karen; Magaye, Ruth; Duke, Trevor

2018-05-01

The Gene Xpert MTB/ RIF assay (Xpert) is used for rapid, simultaneous detection of Mycobacterium tuberculosis (MTB) and rifampicin resistance. This study examined the accuracy of Xpert in children with suspected pulmonary tuberculosis (PTB). Children admitted to Port Moresby General Hospital with suspected PTB were prospectively enrolled between September 2014 and March 2015. They were classified into probable, possible and TB-unlikely groups. Sputum or gastric aspirates were tested by Xpert and smear microscopy; mycobacterial culture was undertaken on a subset. Children were diagnosed with TB on the basis of standard criteria which were used as the primary reference standard. Xpert, smear for acid-fast bacilli (AFB) and the Edwards TB score were compared with the primary reference standard. A total of 93 children ≤14 years with suspected PTB were enrolled; 67 (72%) were classified as probable, 21 (22%) possible and 5 (5.4%) TB-unlikely. Eighty were treated for TB based on the primary reference standard. Xpert was positive in 26/93 (28%) MTB cases overall, including 22/67 (33%) with probable TB and 4/21 (19%) with possible TB. Three (13%) samples identified rifampicin resistance. Xpert confirmed more cases of TB than AFB smear (26 vs 13, p = 0.019). The sensitivity of Xpert, AFB smear and an Edwards TB score of ≥7 was 31% (25/80), 16% (13/80) and 90% (72/80), respectively, and the specificity was 92% (12/13), 100% (13/13) and 31% (4/13), respectively, when compared with the primary reference standard. Xpert sensitivity is sub-optimal and cannot be relied upon for diagnosing TB, although a positive result is confirmatory. A detailed history and examination, standardised clinical criteria, radiographs and available tests remain the most appropriate way of diagnosing TB in children in resource-limited countries. Xpert helps confirm PTB better than AFB smear, and identifies rifampicin resistance. Practical guidelines should be used to identify children who will benefit from an Xpert assay.

Standardization of blood smears prepared in transparent acetate: an alternative method for the microscopic diagnosis of malaria.

PubMed

Mello, Marcia B C; Luz, Francisco C; Leal-Santos, Fabio A; Alves, Eduardo R; Gasquez, Thamires M; Fontes, Cor J F

2014-06-17

Due to students' initial inexperience, slides are frequently broken and blood smears are damaged in microscopy training, leading to the need for their constant replacement. To minimize this problem a method of preparing blood smears on transparent acetate sheets was developed with the goal of implementing appropriate and more readily available teaching resources for the microscopic diagnosis of malaria. Acetate sheets derived from polyester were used to standardize the preparation and staining of thin and thick blood smears on transparent acetate sheets. Thick and thin blood smears were also prepared using the conventional method on glass slides. The staining was conducted using Giemsa staining for the thick and thin smears. Microscopic examination (1,000x) of the thin and thick blood smears prepared on transparent acetate produced high-quality images for both the parasites and the blood cells. The smears showed up on a clear background and with minimal dye precipitation. It was possible to clearly identify the main morphological characteristics of Plasmodium, neutrophils and platelets. After 12 months of storage, there was no change in image quality or evidence of fungal colonization. Preparation of thin and thick blood smears in transparent acetate for the microscopic diagnosis of malaria does not compromise the morphological and staining characteristics of the parasites or blood cells. It is reasonable to predict the applicability of transparent acetate in relevant situations such as the training of qualified professionals for the microscopic diagnosis of malaria and the preparation of positive specimens for competency assessment (quality control) of professionals and services involved in the diagnosis of malaria.

Evaluation of OMNIgene®•SPUTUM-stabilised sputum for long-term transport and Xpert® MTB/RIF testing in Nepal.

PubMed

Maharjan, B; Kelly-Cirino, C D; Weirich, A; Curry, P S; Hoffman, H; Avsar, K; Shrestha, B

2016-12-01

German Nepal TB Project, National Tuberculosis Reference Laboratory, Kathmandu, Nepal. To evaluate whether transporting samples in OMNIgene®•SPUTUM (OM-S) reagent from a peripheral collection site to a central laboratory in Nepal can improve tuberculosis (TB) detection and increase the sensitivity of Xpert® MTB/RIF testing. One hundred sputum samples were split manually. Each portion was assigned to the OM-S group (OM-S added at collection, airline-couriered without cold chain, no other processing required) or the standard-of-care (SOC) group (samples airline-couriered on ice, sodium hydroxide + N-acetyl-L-cysteine processing required at the laboratory). Smear microscopy and Xpert testing were performed. Transport time was 2-13 days. Overall smear results were comparable (respectively 58% and 56% smear-negative results in the OM-S and SOC groups). The rate of smear-positive, Mycobacterium tuberculosis-positive (MTB+) sample detection was identical for both treatment groups, at 95%. More smear-negative MTB+ samples were detected in the OM-S group (17% vs. 13%, P = 0.0655). Sputum samples treated with OM-S can undergo multiday ambient-temperature transport and yield comparable smear and Xpert results to those of SOC samples. Further investigation with larger sample sizes is required to assess whether treating sputum samples with OM-S could increase the sensitivity of Xpert testing in smear-negative samples.

Towards a needle-free diagnosis of malaria: in vivo identification and classification of red and white blood cells containing haemozoin.

PubMed

Burnett, Jennifer L; Carns, Jennifer L; Richards-Kortum, Rebecca

2017-11-07

Optical detection of circulating haemozoin has been suggested as a needle free method to diagnose malaria using in vivo microscopy. Haemozoin is generated within infected red blood cells by the malaria parasite, serving as a highly specific, endogenous biomarker of malaria. However, phagocytosis of haemozoin by white blood cells which persist after the infection is resolved presents the potential for false positive diagnosis; therefore, the focus of this work is to identify a feature of the haemozoin signal to discriminate between infected red blood cells and haemozoin-containing white blood cells. Conventional brightfield microscopy of thin film blood smears was used to analyse haemozoin absorbance signal in vitro. Cell type and parasite maturity were morphologically determined using colocalized DAPI staining. The ability of features to discriminate between infected red blood cells and haemozoin-containing white blood cells was evaluated using images of smears from subjects infected with two species of Plasmodium, Plasmodium yoelii and Plasmodium falciparum. Discriminating features identified by blood smear microscopy were characterized in vivo in P. yoelii-infected mice. Two features of the haemozoin signal, haemozoin diameter and normalized intensity difference, were identified as potential parameters to differentiate infected red blood cells and haemozoin-containing white blood cells. Classification performance was evaluated using the area under the receiver operating characteristic curve, with area under the curve values of 0.89 for the diameter parameter and 0.85 for the intensity parameter when assessed in P. yoelii samples. Similar results were obtained from P. falciparum blood smears, showing an AUC of 0.93 or greater for both classification features. For in vivo investigations, the intensity-based metric was the best classifier, with an AUC of 0.91. This work demonstrates that size and intensity features of haemozoin absorbance signal collected by in vivo microscopy are effective classification metrics to discriminate infected red blood cells from haemozoin-containing white blood cells. This reduces the potential for false positive results associated with optical imaging strategies for in vivo diagnosis of malaria based on the endogenous biomarker haemozoin.

Impact of routine real-time PCR testing of imported malaria over 4 years of implementation in a clinical laboratory.

PubMed

Shokoples, Sandra; Mukhi, Shamir N; Scott, Allison N; Yanow, Stephanie K

2013-06-01

In clinical laboratories, diagnosis of imported malaria is commonly performed by microscopy. However, the volume of specimens is generally low and maintaining proficiency in reading blood smears, particularly at the species level, is challenging in this setting. To address this problem, the Provincial Laboratory for Public Health (ProvLab) in Alberta, Canada, implemented real-time PCR for routine confirmation of all smear-positive samples in the province. Here we report our experience over a 4-year period (2008 to 2012) with this new diagnostic algorithm. While detection of Plasmodium falciparum by microscopy alone was accurate, real-time PCR served as an important adjunct to microscopy for the identification of non-falciparum species. In 18% of cases, the result was reported as non-falciparum or the species could not be identified by microscopy alone, and in all cases, the species was resolved by real-time PCR. In another 4% of cases, the species was misidentified by microscopy. To enhance surveillance for malaria, we integrated our demographic, clinical, and laboratory data into a new system developed by the Canadian Network for Public Health Intelligence, called the Malaria System for Online Surveillance (SOS). Using this application, we characterized our patient populations and travel history to identify risk factors associated with malaria infection abroad.

Use of a pLDH-based dipstick in the diagnostic and therapeutic follow-up of malaria patients in Mali

PubMed Central

2011-01-01

Background Malaria is a major public health problem in Mali and diagnosis is typically based on microscopy. Microscopy requires a well trained technician, a reliable power source, a functioning microscope and adequate supplies. The scarcity of resources of community health centres (CHC) does not allow for such a significant investment in only one aspect of malaria control. In this context, Rapid Diagnostic Tests (RDTs) may improve case management particularly in remote areas. Methods This multicentre study included 725 patients simultaneously screened with OptiMal-IT test and thick smears for malaria parasite detection. While evaluating the therapeutic efficacy of choroquine in 2 study sites, we compared the diagnostic values of thick smear microscopy to OptiMal-IT test applying the WHO 14 days follow-up scheme using samples collected from 344 patients. Results The sensitivity and the specificity of OptiMal-IT compared to thick smear was 97.2% and 95.4%, whereas the positive and negative predictive values were 96.7 and 96.1%, respectively. The percent agreement between the two diagnostic tests was 0.93. The two tests were comparable in detecting malaria at day 0, day 3 and day 14. The only difference was observed at day 7 due to high gametocytemia. Subjectively, health care providers found OptiMal-IT easier to use and store under field conditions. Conclusion OptiMal-IT test revealed similar results when compared to microscopy which is considered the gold standard for malaria diagnostics. The test was found to have a short processing time and was easier to use. These advantages may improve malaria case management by providing a diagnostic and drug efficacy follow-up tool to peripheral health centres with limited resources. PMID:22114867

Recent advances in tuberculosis diagnostics in resource-limited settings.

PubMed

Seki, Mitsuko; Kim, Chang-Ki; Hayakawa, Satoshi; Mitarai, Satoshi

2018-04-19

Smear-negative and drug-resistant cases of tuberculosis (TB) disease necessitate the development of new diagnostic methods, especially in resource-limited settings. To improve the current TB situations, sensitive and specific TB point-of-care tests (POCTs) should be developed. This review addresses the current status of TB, novel diagnostic methodologies for TB, and the impact of those new diagnostics on TB control in such situations. Moreover, the perspective of TB management based on laboratory examinations is described. Smear microscopy with sputum samples is the only laboratory examination available in many resource-limited settings and is still used globally. Several nucleic acid amplification tests (NATs) have been developed. The World Health Organization (WHO) endorsed novel diagnostics based on NATs and updated their definition of a bacteriologically confirmed case requiring the biological specimen to be positive by smear microscopy, culture, or the WHO-recommended rapid diagnostic protocols. The use of new diagnostics increased the number of bacteriologically confirmed TB cases. Novel diagnostics are now available, but their sensitivity is still lower than that of conventional liquid culture method. To address the increasing incidence of TB, more resources including novel diagnostics as POCTs with higher sensitivity must be allocated to healthcare systems.

Scanning electron microscopy (SEM) evaluation of sealing ability of MTA and EndoSequence as root-end filling materials with chitosan and carboxymethyl chitosan (CMC) as retrograde smear layer removing agents.

PubMed

Nagesh, Bolla; Jeevani, Eppala; Sujana, Varri; Damaraju, Bharagavi; Sreeha, Kaluvakolanu; Ramesh, Penumaka

2016-01-01

The purpose of this study was to evaluate the sealing ability of mineral trioxide aggregate (MTA) and EndoSequence with chitosan and carboxymethyl chitosan (CMC) as retrograde smear layer removing agents using scanning electron microscopy (SEM). Forty human single rooted teeth were taken. Crowns were decoronated and canals were obturated. Apically roots were resected and retrograde cavities were done. Based on the type of retrograde material placed and the type of smear layer removal agent used for retrograde cavities, they were divided into four groups (N = 10): Group I chitosan with EndoSequence, group II chitosan with MTA, group III CMC with EndoSequence, and Group IV CMC with MTA. All the samples were longitudinally sectioned, and the SEM analysis was done for marginal adaptation. Kruskal-Wallis and Mann-Witney analysis tests. SEM images showed the presence of less gaps in group III, i.e., CMC with EndoSequence when compared to other groups with statistically significant difference. Within the limited scope of this study, it was concluded that EndoSequence as retrograde material showed better marginal sealing ability.

Diagnosis of smear-negative pulmonary tuberculosis based on clinical signs in the Republic of Congo.

PubMed

Linguissi, Laure Stella Ghoma; Vouvoungui, Christevy Jeannhey; Poulain, Pierre; Essassa, Gaston Bango; Kwedi, Sylvie; Ntoumi, Francine

2015-12-18

The diagnosis of pulmonary tuberculosis (PTB) and smear-negative pulmonary tuberculosis (SNPT) in resource-limited countries is often solely based on clinical signs, chest X-ray radiography and sputum smear microscopy. We investigated currently used methods for the routine diagnosis of SNPT in the Republic of Congo (RoC) among TB suspected patients. The specific case of HIV positive patients was also studied. A cross-sectional study was conducted at the anti-tuberculosis center (CAT) of Brazzaville, RoC. Tuberculosis suspects were examined for physical signs of TB. Clinical signs, results from sputum smear microscopy, tuberculin skin test (TST) and chest X-ray were recorded. Of the 772 enrolled participants, 372 were diagnosed PTB. Cough was a common symptom for PTB and no PTB patients. Pale skin, positive TST, weight loss and chest X-ray with abnormalities compatible with PTB (PTB-CXR) were significant indicators of PTB. Thirty-six percent of PTB patients were diagnosed SNPT. This category of patients presented less persistent cough and less PTB-CXR. Anorexia and asthenia were significant indicators of SNPT. In the case of HIV+ patients, 57% were SNPT with anorexia, asthenia and shorter cough being strong indicators of SNPT. Chest X-ray abnormalities, weight loss, pale skin and positive TST were significant indicators of PTB. Anorexia and asthenia showed good diagnostic performance for SNPT, which deserve to be recommended as index indicators of SNPT diagnosis. Duration of cough is also a relevant indicator, especially for HIV+ patients.

Effects of modifying the World Health Organization standard operating procedures for malaria microscopy to improve surveillance in resource poor settings.

PubMed

Fernando, Sumadhya D; Ihalamulla, Ratnasiri L; Wickremasinghe, Renu; de Silva, Nipun L; Thilakarathne, Janani H; Wijeyaratne, Pandu; Premaratne, Risintha G

2014-03-15

Individuals with fever are screened for malaria in specially-established malaria diagnostic laboratories set up in rural hospitals in the Northern and Eastern Provinces of Sri Lanka. Large numbers of blood smears negative for malaria parasites are being screened daily. Good quality smears are essential to maintain a high diagnostic competency among the technical staff. The modifications made to the World Health Organization (WHO) standard operating procedures to improve the quality of smears have been studied. A blinded, controlled, interventional study was conducted in 22 intervention and 21 control malaria diagnostic laboratories. Changes were made to the WHO standard operating procedure protocols to prepare, stain and examine blood smears for malaria parasite detection which were implemented in intervention laboratories. These included wipe-cleaning slides, preparing both thick and thin smears on the same slide, reversing the order of collecting blood for thick and thin smears, dry fixing thick smear for 20-25 minutes under table lamp, polishing the edge of spreader slide with sand paper and fixing the thin smear with methanol if not stained within four hours. Parameters with respect to quality of the smear as per WHO criteria were studied using randomly selected slides, and time taken for the report to be issued was recorded in both groups before and after the intervention. There were no significant differences observed in the parameters studied at baseline between the two groups or pre and post intervention in the control group. In the intervention group streak formation in thin smears was reduced from 29.4% to 5.0%. The average fixing time of thick smears was reduced from 2.4 hours to 20 minutes. Inappropriate thickness of thick smears reduced from 18.3% to 1.5%. Overall quality of thick smears and thin smears increased from 76.1% to 98.0% and 81.7% to 87.0%, respectively. The quality of slides bearing both thick and thin smears increased from 60.0% to 87.0%. New protocols with amendments to the WHO standard technical procedures ensure that good quality blood smears are prepared rapidly to diagnose malaria and the time required to issue the reports was reduced.

Safety assessment in primary Mycobacterium tuberculosis smear microscopy centres in Blantyre Malawi: a facility based cross sectional survey.

PubMed

Majamanda, J; Ndhlovu, P; Shawa, I T

2013-12-01

Tuberculosis (TB) is caused by Mycobacterium tuberculosis and is transmitted mainly through aerosolization of infected sputum which puts laboratory workers at risk in spite of the laboratory workers' risk of infection being at 3 to 9 times higher than the general public. Laboratory safety should therefore be prioritized and optimized to provide sufficient safety to laboratory workers. To assess the safety for the laboratory workers in TB primary microscopy centres in Blantyre urban. TB primary microscopy centers in Blantyre urban were assessed in aspects of equipment availability, facility layout, and work practice, using a standardized WHO/AFRO ISO 15189 checklist for the developing countries which sets the minimum safety score at ≥80%. Each center was graded according to the score it earned upon assessment. Only one (1) microscopy center out nine (9) reached the minimum safety requirement. Four (4) centers were awarded 1 star level, four (4) centers were awarded 2 star level and only one (1) center was awarded 3 star level. In Blantyre urban, 89% of the Tuberculosis microscopy centers are failing to provide the minimum safety to the laboratory workers. Government and other stake holders should be committed in addressing the safety challenges of TB microscopy centres in the country to ensure safety for the laboratory workers. It is recommended that the study be conducted at the regional or national level for both public and private laboratories in order to have a general picture of safety in Tb microscopy centres possibly across the country.

Incidence of smear-positive tuberculosis in Dabat, northern Ethiopia.

PubMed

Tadesse, T; Demissie, M; Berhane, Y; Kebede, Y; Abebe, M

2013-05-01

To determine the incidence of smear-positive tuberculosis (TB) in Dabat District, northern Ethiopia. Using a population-based longitudinal design, a TB surveillance system was initiated among 46,165 residents at the Dabat Health and Demographic Surveillance System site. Trained field workers visited each household every third month and interviewed all individuals aged ≥14 years using a uniform questionnaire to detect suspected cases of TB (cough ≥15 days), at which time two sputum (spot-morning) samples were collected for smear microscopy. A total of 281,820 person-months were observed during the 1-year period, which generated 74 smear-positive TB cases. The incidence of smear-positive TB was calculated at 311 per 100,000 person-years (95%CI 240-382). Higher rates were observed among females (incidence rate ratio [IRR] 2.08, 95%CI 1.24-3.52), persons with no schooling (IRR 2.74, 95%CI 1.11-6.78) and urban residents (IRR 2.39, 95%CI 1.39-4.12). The incidence of smear-positive TB is high in Dabat District, suggesting a high risk of transmission in the communities. TB control programmes thus need to improve case-finding mechanisms at the community level in Ethiopia, with greater emphasis on risk groups.

Number of sputum specimens during treatment follow-up of tuberculosis patients: two or one?

PubMed Central

Kumar, A. M. V.; Claassens, M.; Banurekha, V. V.; Gomathi, N. S.; Venkatesan, P.; Swaminathan, S.

2013-01-01

Setting: National Institute for Research in Tuberculosis clinics in Chennai and Madurai, India. Objective: To examine the pattern of serial smears (negative-negative [NN], negative-positive [NP], positive-negative [PN], positive-positive [PP]) during treatment follow-up of culture-confirmed new smear-positive tuberculosis (TB) patients, and the proportion of culture-negatives in each category. Design: We reviewed the records and extracted follow-up smear (fluorescent microscopy) and culture (Löwenstein-Jensen) results of patients enrolled in clinical trials from January 2000 to August 2012 and treated with the Category I regimen (2EHRZ3/4HR3). Data entry and analysis were performed using EpiData. Results: Among 520 patients (176 infected with the human immunodeficiency virus), the proportions of culture-negative patients with NN, discordant (PN or NP) and PP patterns were approximately 98%, 80% and 40%, respectively. The smear-positive culture-negative phenomenon was more frequent in follow-up smear results graded 1+, followed by 2+ and 3+. Conclusion: There is justification for discontinuing the examination of second specimens during treatment follow-up among TB patients. However, a positive result on the first smear needs to be confirmed by a second positive result before making clinical management decisions. The World Health Organization may need to reconsider its recommendation on this issue. PMID:26393051

The utility of Xpert MTB/RIF performed on bronchial washings obtained in patients with suspected pulmonary tuberculosis in a high prevalence setting.

PubMed

Barnard, Dewald A; Irusen, Elvis M; Bruwer, Johannes W; Plekker, Danté; Whitelaw, Andrew C; Deetlefs, Jacobus D; Koegelenberg, Coenraad F N

2015-09-16

Xpert MTB/RIF has been shown to have a superior sensitivity to microscopy for acid fast bacilli (AFB) in sputum and has been recommended as a standard first line investigation for pulmonary tuberculosis (PTB). Bronchoscopy is a valuable tool in diagnosing PTB in sputum negative patients. There is limited data on the utility of Xpert MTB/RIF performed on bronchial lavage specimens. Our aim was to evaluate the diagnostic efficiency of Xpert MTB/RIF performed on bronchial washings in sputum scarce/negative patients with suspected PTB. All patients with a clinical and radiological suspicion of PTB who underwent bronchoscopy between January 2013 and April 2014 were included. The diagnostic efficiencies of Xpert MTB/RIF and microscopy for AFB were compared to culture for Mycobacterium tuberculosis. Thirty nine of 112 patients were diagnosed with culture-positive PTB. Xpert MTB/RIF was positive in 36/39 with a sensitivity of 92.3% (95% CI 78-98%) for PTB, which was superior to that of smear microscopy (41%; 95% CI 26.0-57.8%, p = 0.005). The specificities of Xpert MTB/RIF and smear microscopy were 87.7% (95% CI 77.4-93.9%) and 98.6% (95% CI 91.6%-99.9%) respectively. Xpert MTB/RIF had a positive predictive value of 80% (95% CI; 65-89.9%) and negative predictive value of 95.5% (95% CI 86.6-98.8%). 3/9 patients with Xpert MTB/RIF positive culture negative results were treated for PTB based on clinical and radiological findings. Xpert MTB/RIF has a higher sensitivity than smear microscopy and similar specificity for the immediate confirmation of PTB in specimens obtained by bronchial washing, and should be utilised in patients with a high suspicion of pulmonary tuberculosis.

Performance of Xpert® MTB/RIF among tuberculosis outpatients in Lilongwe, Malawi.

PubMed

Chikaonda, Tarsizio; Nguluwe, Nelson; Barnett, Brian; Gokhale, Runa H; Krysiak, Robert; Thengolose, Isaac; Rosenberg, Nora E; Stanley, Christopher; Mpunga, James; Hoffman, Irving F; Hosseinipour, Mina; Scott, Lesley; Stevens, Wendy

2017-01-01

Xpert ® MTB/RIF is a molecular test for the detection of Mycobacterium tuberculosis and rifampicin resistance. It is considered to be a great advance over smear microscopy and culture. However, there is very little information regarding the performance characteristics of Xpert MTB/RIF in Malawi. We aimed to evaluate the performance of Xpert MTB/RIF in a Malawian setting. Stored sputum pellets were processed on Xpert MTB/RIF between June 2012 and May 2014. Results were compared to mycobacteria growth indicator tube and Löwenstein-Jensen cultures, LED fluorescent microscopy and GenoType ® MTBDR plus assay. Rifampicin resistance was confirmed by DNA sequencing. Of the 348 specimens with valid Xpert MTB/RIF results, 129/348 (37%) were smear-positive and 198/348 (57%) were culture-positive. Xpert MTB/RIF demonstrated a sensitivity of 93.8% (95% CI 89.4% - 96.8%) and specificity of 97.4% (95% CI 93.5% - 99.3%), with a positive predictive value of 97.8% (95% CI 94.6% - 99.4%) and a negative predictive value of 92.6% (95% CI 87.4% - 96.1%). Xpert MTB/RIF correctly identified 185/186 (99.5%) rifampicin-sensitive and 2/2 (100%) rifampicin-resistant M. tuberculosis strains. Mutations were not detected by sequencing in one isolate which was rifampicin resistant on Xpert MTB/RIF but sensitive on MTBDR plus . Four non-tuberculous mycobacteria grew from four smear-negative specimens, namely, M. avium ( n = 1) and M. intracellulare ( n = 3). No cross-reactivity was observed with any of the non-tuberculous mycobacteria when using Xpert MTB/RIF. When fully implemented, Xpert MTB/RIF may have an impact on patient care in Malawi. The increased diagnostic yield of Xpert MTB/RIF over smear microscopy can increase laboratory-confirmed tuberculosis detection and ensure that treatment is given to appropriate individuals or groups.

Performance of Xpert® MTB/RIF among tuberculosis outpatients in Lilongwe, Malawi

PubMed Central

Nguluwe, Nelson; Gokhale, Runa H.; Krysiak, Robert; Thengolose, Isaac; Rosenberg, Nora E.; Stanley, Christopher; Mpunga, James; Hoffman, Irving F.; Hosseinipour, Mina; Scott, Lesley; Stevens, Wendy

2017-01-01

Background Xpert® MTB/RIF is a molecular test for the detection of Mycobacterium tuberculosis and rifampicin resistance. It is considered to be a great advance over smear microscopy and culture. However, there is very little information regarding the performance characteristics of Xpert MTB/RIF in Malawi. Objective We aimed to evaluate the performance of Xpert MTB/RIF in a Malawian setting. Methods Stored sputum pellets were processed on Xpert MTB/RIF between June 2012 and May 2014. Results were compared to mycobacteria growth indicator tube and Löwenstein-Jensen cultures, LED fluorescent microscopy and GenoType® MTBDRplus assay. Rifampicin resistance was confirmed by DNA sequencing. Results Of the 348 specimens with valid Xpert MTB/RIF results, 129/348 (37%) were smear-positive and 198/348 (57%) were culture-positive. Xpert MTB/RIF demonstrated a sensitivity of 93.8% (95% CI 89.4% – 96.8%) and specificity of 97.4% (95% CI 93.5% – 99.3%), with a positive predictive value of 97.8% (95% CI 94.6% – 99.4%) and a negative predictive value of 92.6% (95% CI 87.4% – 96.1%). Xpert MTB/RIF correctly identified 185/186 (99.5%) rifampicin-sensitive and 2/2 (100%) rifampicin-resistant M. tuberculosis strains. Mutations were not detected by sequencing in one isolate which was rifampicin resistant on Xpert MTB/RIF but sensitive on MTBDRplus. Four non-tuberculous mycobacteria grew from four smear-negative specimens, namely, M. avium (n = 1) and M. intracellulare (n = 3). No cross-reactivity was observed with any of the non-tuberculous mycobacteria when using Xpert MTB/RIF. Conclusion When fully implemented, Xpert MTB/RIF may have an impact on patient care in Malawi. The increased diagnostic yield of Xpert MTB/RIF over smear microscopy can increase laboratory-confirmed tuberculosis detection and ensure that treatment is given to appropriate individuals or groups. PMID:28879160

Evaluation of ethylenediaminetetraacetic acid (EDTA) solution and gel for smear layer removal.

PubMed

Dotto, Sidney Ricardo; Travassos, Rosana Maria Coelho; de Oliveira, Elias Pandonor Motcy; Machado, Manoel Eduardo de Lima; Martins, José Luiz

2007-08-01

The purpose of this in vitro study was to compare the efficacy of 24% ethylenediaminetetraacetic acid (EDTA) gel and 17% EDTA solution in cleaning dentine walls after root canal instrumentation. Thirty human canine teeth were divided into three groups of 10 teeth each. In Group 1, 1% sodium hypochlorite was used as the irrigating solution; in Group 2, 1% sodium hypochlorite was used with 17% EDTA solution; and in Group 3, 1% sodium hypochlorite was used with 24% EDTA gel. The presence of a smear layer was analysed after instrumentation using scanning electron microscopy. The Kruskal-Wallis test revealed a statistical difference (P < 0.05) between Groups 1 and 2, and also between Groups 1 and 3. No difference was observed between Groups 2 and 3 (P > 0.05). The results indicate that 1% sodium hypochlorite alone does not remove the smear layer and that there was no statistical difference between EDTA gel and EDTA solution in smear layer removal.

Effect of Aquatine Endodontic Cleanser on smear layer removal in the root canals of ex vivo human teeth.

PubMed

Garcia, Faustino; Murray, Peter E; Garcia-Godoy, Franklin; Namerow, Kenneth N

2010-01-01

The purpose of this study was to measure and compare the root canal cleanliness and smear layer removal effectiveness of Aquatine Endodontic Cleanser (Aquatine EC) when used as an endodontic irrigating solution in comparison with 6% sodium hypochlorite (NaOCl). Forty-five human teeth were randomly allocated to five treatment groups; the pulp chamber was accessed, cleaned, and shaped by using ProTaper and ProFile rotary instrumentation to an ISO size #40. The teeth were then processed for scanning electron microscopy, and the root canal cleanliness and removal of smear layer were examined. The most effective removal of smear layer occurred with Aquatine EC and NaOCl, both with a rinse of EDTA. Aquatine EC appears to be the first hypochlorous acid approved by the FDA to be a possible alternative to the use of NaOCl as an intracanal irrigant. Further research is needed to identify safer and more effective alternatives to the use of NaOCl irrigation in endodontics.

Comparative Study of Malaria Prevalence among Travellers in Nigeria (West Africa) Using Slide Microscopy and a Rapid Diagnosis Test.

PubMed

Dougnon, T V; Bankole, H S; Hounmanou, Y M G; Echebiri, S; Atchade, P; Mohammed, J

2015-01-01

Malaria is a major disease in Africa and leads to various public health problems. A study was carried out at the Aviation Medical Clinic Laboratory, Murtala Mohammed Airport, Ikeja, Lagos State, Nigeria, in 2014. The work aimed to determine the prevalence of malaria among patients attending the laboratory. Blood samples were therefore collected from 51 patients and subjected to both blood smear microscopy and a rapid immunochromatographic diagnostic test (SD BIOLINE Malaria Ag) for detection of, respectively, malaria parasites and antigens. At the end of the study, 22% of the patients were detected positive by the microscopic examination while 9.8% were tested positive when using SD BIOLINE Malaria Ag. The outcomes of the study show a high prevalence of malaria at the airport. This represents a serious risk factor leading to a high likelihood of spread and occurrence of malaria in other countries including Western countries whereby the disease is nonendemic. It also pointed out that the blood smear microscopy seems to be better than Rapid Diagnosis Test (RDT) for malaria diagnosis.

Active case finding of undetected tuberculosis among chronic coughers in a slum setting in Kampala, Uganda

PubMed Central

Sekandi, J. N.; Neuhauser, D.; Smyth, K.; Whalen, C. C.

2010-01-01

SUMMARY SETTING Kisenyi slum in peri-urban Kampala, Uganda. OBJECTIVES Using chronic cough (≥2 weeks) inquiry as a screening tool to identify undetected smear-positive tuberculosis (TB) cases and to describe the characteristics of smear-positive TB cases detected by active case finding. DESIGN A house-to-house survey was conducted in five randomly selected villages in Kampala between June and August 2005. A sample of households was visited; adults aged ≥15 years were consecutively interviewed to identify those with chronic cough. Three sputum specimens were collected and examined by smear microscopy. RESULTS Among 930 individuals, we identified 189 (20%) chronic coughers. Of these, we found 33 (18%) undiagnosed smear-positive cases. The newly detected cases had an even sex distribution (P = 0.47), a median age of 30 years, a median cough duration of 1 month and 55% had acid-fast bacilli 1+ sputum smear grade. CONCLUSION These findings suggest that active case finding could supplement DOTS to yield additional smear-positive TB cases, lead to early diagnosis and thus shorten the duration of infectiousness before effective chemotherapy is initiated. In communities such as Kisenyi, this is a feasible strategy that may prove useful for TB control, but its cost-effectiveness needs to be evaluated. Early health care seeking for cough should be emphasized. PMID:19335958

Comparative study of two protocols for quantitative image-analysis of serotonin transporter clustering in lymphocytes, a putative biomarker of therapeutic efficacy in major depression.

PubMed

Romay-Tallon, Raquel; Rivera-Baltanas, Tania; Allen, Josh; Olivares, Jose M; Kalynchuk, Lisa E; Caruncho, Hector J

2017-01-01

The pattern of serotonin transporter clustering on the plasma membrane of lymphocytes extracted from human whole blood samples has been identified as a putative biomarker of therapeutic efficacy in major depression. Here we evaluated the possibility of performing a similar analysis using blood smears obtained from rats, and from control human subjects and depression patients. We hypothesized that we could optimize a protocol to make the analysis of serotonin protein clustering in blood smears comparable to the analysis of serotonin protein clustering using isolated lymphocytes. Our data indicate that blood smears require a longer fixation time and longer times of incubation with primary and secondary antibodies. In addition, one needs to optimize the image analysis settings for the analysis of smears. When these steps are followed, the quantitative analysis of both the number and size of serotonin transporter clusters on the plasma membrane of lymphocytes is similar using both blood smears and isolated lymphocytes. The development of this novel protocol will greatly facilitate the collection of appropriate samples by eliminating the necessity and cost of specialized personnel for drawing blood samples, and by being a less invasive procedure. Therefore, this protocol will help us advance the validation of membrane protein clustering in lymphocytes as a biomarker of therapeutic efficacy in major depression, and bring it closer to its clinical application.

Accuracy and impact of Xpert MTB/RIF for the diagnosis of smear-negative or sputum-scarce tuberculosis using bronchoalveolar lavage fluid.

PubMed

Theron, Grant; Peter, Jonny; Meldau, Richard; Khalfey, Hoosain; Gina, Phindile; Matinyena, Brian; Lenders, Laura; Calligaro, Gregory; Allwood, Brian; Symons, Gregory; Govender, Ureshnie; Setshedi, Mashiko; Dheda, Keertan

2013-11-01

The accuracy and impact of new tuberculosis (TB) tests, such as Xpert MTB/RIF, when performed on bronchoalveolar lavage fluid (BALF) obtained from patients with sputum-scarce or smear-negative TB is unclear. South African patients with suspected pulmonary TB (n=160) who were sputum-scarce or smear-negative underwent bronchoscopy. MTB/RIF was performed on uncentrifuged BALF (1 ml) and/or a resuspended pellet of centrifuged BALF (∼10 ml). Time to TB detection and anti-TB treatment initiation were compared between phase one, when MTB/RIF was performed as a research tool, and phase two, when it was used for patient management. 27 of 154 patients with complete data had culture-confirmed TB. Of these, a significantly lower proportion were detected by smear microscopy compared with MTB/RIF (58%, 95% CI 39% to 75% versus 93%, 77% to 98%; p<0.001). Of the 127 patients who were culture negative, 96% (91% to 98%) were MTB/RIF negative. When phase two was compared with phase one, MTB/RIF reduced the median days to TB detection (29 (18-41) to 0 (0-0); p<0.001). However, more patients initiated empirical therapy (absence of a positive test in those commencing treatment) in phase one versus phase two (79% (11/14) versus 28% (10/25); p=0.026). Consequently, there was no detectable difference in the overall proportion of patients initiating treatment (26% (17/67; 17% to 37%) versus 36% (26/73; 26% to 47%); p=0.196) or the days to treatment initiation (10 (1-49) versus 7 (0-21); p=0.330). BALF centrifugation, HIV coinfection and a second MTB/RIF did not result in detectable changes in accuracy. MTB/RIF detected TB cases more accurately and more rapidly than smear microscopy and significantly reduced the rate of empirical treatment.

Accuracy and impact of Xpert MTB/RIF for the diagnosis of smear-negative or sputum-scarce tuberculosis using bronchoalveolar lavage fluid

PubMed Central

Theron, Grant; Peter, Jonny; Meldau, Richard; Khalfey, Hoosain; Gina, Phindile; Matinyena, Brian; Lenders, Laura; Calligaro, Gregory; Allwood, Brian; Symons, Gregory; Govender, Ureshnie; Setshedi, Mashiko; Dheda, Keertan

2017-01-01

Rationale The accuracy and impact of new tuberculosis (TB) tests, such as Xpert MTB/RIF, when performed on bronchoalveolar lavage fluid (BALF) obtained from patients with sputum-scarce or smear-negative TB is unclear. Methods South African patients with suspected pulmonary TB (n=160) who were sputum-scarce or smear-negative underwent bronchoscopy. MTB/RIF was performed on uncentrifuged BALF (1 ml) and/or a resuspended pellet of centrifuged BALF (~10 ml). Time to TB detection and anti-TB treatment initiation were compared between phase one, when MTB/RIF was performed as a research tool, and phase two, when it was used for patient management. Results 27 of 154 patients with complete data had culture-confirmed TB. Of these, a significantly lower proportion were detected by smear microscopy compared with MTB/RIF (58%, 95% CI 39% to 75% versus 93%, 77% to 98%; p<0.001). Of the 127 patients who were culture negative, 96% (91% to 98%) were MTB/RIF negative. When phase two was compared with phase one, MTB/RIF reduced the median days to TB detection (29 (18–41) to 0 (0–0); p<0.001). However, more patients initiated empirical therapy (absence of a positive test in those commencing treatment) in phase one versus phase two (79% (11/14) versus 28% (10/25); p=0.026). Consequently, there was no detectable difference in the overall proportion of patients initiating treatment (26% (17/67; 17% to 37%) versus 36% (26/73; 26% to 47%); p=0.196) or the days to treatment initiation (10 (1–49) versus 7 (0–21); p=0.330). BALF centrifugation, HIV coinfection and a second MTB/RIF did not result in detectable changes in accuracy. Conclusions MTB/RIF detected TB cases more accurately and more rapidly than smear microscopy and significantly reduced the rate of empirical treatment. PMID:23811536

Burden of tuberculosis in intensive care units in Cape Town, South Africa, and assessment of the accuracy and effect on patient outcomes of the Xpert MTB/RIF test on tracheal aspirate samples for diagnosis of pulmonary tuberculosis: a prospective burden of disease study with a nested randomised controlled trial.

PubMed

Calligaro, Gregory L; Theron, Grant; Khalfey, Hoosain; Peter, Jonathan; Meldau, Richard; Matinyenya, Brian; Davids, Malika; Smith, Liezel; Pooran, Anil; Lesosky, Maia; Esmail, Aliasgar; Miller, Malcolm G; Piercy, Jenna; Michell, Lancelot; Dawson, Rodney; Raine, Richard I; Joubert, Ivan; Dheda, Keertan

2015-08-01

There are few prospective data about the incidence and mortality associated with pulmonary tuberculosis in intensive care units (ICUs), and none on the accuracy and clinical effect of the Xpert-MTB/RIF assay in this setting. We aimed to measure the frequency of culture-positive tuberculosis in ICUs in Cape Town, South Africa and to assess the performance and effect on patient outcomes of Xpert MTB/RIF versus smear microscopy for diagnosis of tuberculosis. We did a prospective burden of disease study with a randomised controlled substudy at the ICUs of four hospitals in Cape Town. Mechanically ventilated adults (≥18 years) with suspected pulmonary tuberculosis admitted between Aug 1, 2010, and July 31, 2013 (irrespective of the reason for admission), were prospectively investigated by culture, and by Xpert-MTB/RIF testing or smear microscopy, of tracheal aspirate samples. In the substudy, patients were randomly assigned (1:1), via a computer-generated allocation list, to smear microscopy or Xpert MTB/RIF. Participants, caregivers, and outcome assessors were not masked to group assignment. Only the laboratory staff were blinded to the clinical details of the participants. In November, 2012, Xpert MTB/RIF was adopted as the initial diagnostic test for respiratory samples in Western Cape province. Thereafter, patients received Xpert MTB/MIF and culture as standard of care. For the whole study cohort, the primary outcome was the frequency of bacteriologically confirmed tuberculosis. The primary endpoint of the randomised substudy was the proportion of culture-positive patients on treatment at 48 h after enrolment. The randomised substudy is registered with ClinicalTrials.gov, number NCT01530568. We investigated 341 patients for suspected pulmonary tuberculosis out of a total of 2309 ICU admissions. 46 (15%) of 317 patients included in the final analysis had a positive test for tuberculosis (Xpert MTB/RIF or culture). Culture-positive patients who failed to initiate treatment (adjusted HR 4·49, 95% CI 1·45-13·89) or who received inotropes (4·33, 1·49-12·60) were more likely to die. However, tuberculosis status was not associated with 28-day or 90-day mortality. In the substudy, we randomly assigned 115 patients to smear microscopy and 111 to Xpert MTB/RIF. Smear microscopy detected six (43%) of 14 culture-positive patients, and Xpert MTB/RIF detected 11 (100%) of 11 culture-positive patients (p=0·002). The proportion of culture-positive patients on treatment at 48 h was higher in the Xpert MTB/RIF group than in the smear microscopy group (11 [92%] of 12 vs nine [53%] of 17; p=0·043), although use of Xpert MTB/RIF had no effect on mortality or other patient outcomes. Tuberculosis is fairly common in ICUs in high-burden settings, and clinicians should screen and test patients for tuberculosis with Xpert MTB/RIF where available. This test improves diagnostic yield and rates of treatment initiation, and reduces unnecessary treatment, but might not increase the total number of patients on treatment when empirical treatment is widely used. A suspected diagnosis of pulmonary tuberculosis should not exclude patients from ICU care in resource-limited settings because mortality is unaffected by the presence of this disease. European and Developing Countries Clinical Trials Partnership, South African Medical Research Council, and the Discovery Foundation. Copyright © 2015 Elsevier Ltd. All rights reserved.

Evaluation of GeneXpert MTB/RIF for detection of pulmonary tuberculosis at peripheral tuberculosis clinics.

PubMed

Shao, Yan; Peng, Hong; Chen, Cheng; Zhu, Tao; Ji, Ming; Jiang, Wei; Zhu, Wei; Zhai, Xiang Jun; Lu, Wei

2017-04-01

Tuberculosis is one of the most common infectious diseases in China, while delayed patient finding obstructed disease control, especially for smear-negative patients. The current study was undertaken to evaluate the diagnostic accuracy of GeneXpert MTB/RIF compared with conventional methods in the detection of pulmonary tuberculosis patients. A total of 295 spot sputum samples from confirmed pulmonary tuberculosis patients were evaluated from September 2014 to June 2015. Each sample was examined by acid-fast bacillus smear microscopy, culture and GeneXpert MTB/RIF. The sputum culture on Löwenstein-Jensen (L-J) was considered as the gold-standard. After testing by smear, 68.81% (203/295) was negative and 31.19% (92/295) was positive. As the gold-standard, L-J culture detected 37.97% (112/295) positive of all specimens, while the positivity for GeneXpert MTB/RIF was 46.44% (137/295). Compared with L-J culture, the combined sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for GeneXpert MTB/RIF were 94.64%, 82.97%, 77.37% and 96.18% respectively. For smear-negative specimens, the sensitivity, specificity, PPV and NPV for GeneXpert MTB/RIF were 96.00%, 83.05%, 44.44% and 99.32%; while for smear-positive specimens, the corresponding accuracy values were 94.25%, 80.00%, 98.80% and 44.44%. The findings of study indicated that GeneXpert MTB/RIF assay demonstrated a high sensitivity in detecting Mycobacterium tuberculosis compared to smear method and a high NPV among smear negative patients. Copyright © 2017 Elsevier Ltd. All rights reserved.

Optimization of liquid scintillation measurements applied to smears and aqueous samples collected in industrial environments

NASA Astrophysics Data System (ADS)

Chapon, Arnaud; Pigrée, Gilbert; Putmans, Valérie; Rogel, Gwendal

Search for low-energy β contaminations in industrial environments requires using Liquid Scintillation Counting. This indirect measurement method supposes a fine control from sampling to measurement itself. Thus, in this paper, we focus on the definition of a measurement method, as generic as possible, for both smears and aqueous samples' characterization. That includes choice of consumables, sampling methods, optimization of counting parameters and definition of energy windows, using the maximization of a Figure of Merit. Detection limits are then calculated considering these optimized parameters. For this purpose, we used PerkinElmer Tri-Carb counters. Nevertheless, except those relative to some parameters specific to PerkinElmer, most of the results presented here can be extended to other counters.

Effect of aquatine endodontic cleanser on smear layer removal in the root canals of ex vivo human teeth

PubMed Central

GARCIA, Faustino; MURRAY, Peter E.; GARCIA-GODOY, Franklin; NAMEROW, Kenneth N.

2010-01-01

Objectives The purpose of this study was to measure and compare the root canal cleanliness and smear layer removal effectiveness of Aquatine Endodontic Cleanser (Aquatine EC) when used as an endodontic irrigating solution in comparison with 6% sodium hypochlorite (NaOCl). Material and Methods Forty-five human teeth were randomly allocated to five treatment groups; the pulp chamber was accessed, cleaned, and shaped by using ProTaper and ProFile rotary instrumentation to an ISO size #40. The teeth were then processed for scanning electron microscopy, and the root canal cleanliness and removal of smear layer were examined. Results The most effective removal of smear layer occurred with Aquatine EC and NaOCl, both with a rinse of EDTA. Conclusions Aquatine EC appears to be the first hypochlorous acid approved by the FDA to be a possible alternative to the use of NaOCl as an intracanal irrigant. Further research is needed to identify safer and more effective alternatives to the use of NaOCl irrigation in endodontics. PMID:20835577

42 CFR 493.19 - Provider-performed microscopy (PPM) procedures.

Code of Federal Regulations, 2010 CFR

2010-10-01

... HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY REQUIREMENTS General Provisions § 493... mount preparations for the presence or absence of bacteria, fungi, parasites, and human cellular... examinations. (7) Nasal smears for granulocytes. (8) Fecal leukocyte examinations. (9) Qualitative semen...

42 CFR 493.19 - Provider-performed microscopy (PPM) procedures.

Code of Federal Regulations, 2011 CFR

2011-10-01

... HUMAN SERVICES (CONTINUED) STANDARDS AND CERTIFICATION LABORATORY REQUIREMENTS General Provisions § 493... mount preparations for the presence or absence of bacteria, fungi, parasites, and human cellular... examinations. (7) Nasal smears for granulocytes. (8) Fecal leukocyte examinations. (9) Qualitative semen...

Efficacy of Sodium Hypochlorite Activated With Laser in Intracanal Smear Layer Removal: An SEM Study

PubMed Central

Shahriari, Shahriar; Kasraei, Shahin; Roshanaei, Ghodratollah; Karkeabadi, Hamed; Davanloo, Hossein

2017-01-01

Introduction: The purpose of the present study was to evaluate the different concentrations of sodium hypochlorite activated with laser in removing of the smear layer in the apical, middle, and coronal segments of root canal walls by scanning electron microscopy analysis. Methods: Sixty single-rooted human mandibular teeth were decoronated to a standardized length. The samples were prepared by using Race rotary system to size 40, 0.04 taper and divided into 4 equal groups (n = 15). Group 1, irrigated with EDTA 17% and 5.25% NaOCl, groups 2, 3 and 4, 1%, 2.5%, and 5% NaOCl activated with Nd:YAG laser, respectively. Teeth were split longitudinally and subjected to scanning electron microscope (SEM). Data were analyzed by Kruskal-Wallis, Mann-Whitney tests. P value of <0.05 was considered statistically significant. Results: Five percent NaOCl LAI (laser-activated irrigation) showed best smear layer removal in test groups and the difference was statistically significant (P < 0.001). Control group (EDTA 17% and 5.25% NaOCl irrigation) showed significantly better outcomes in comparative with test groups (P < 0.001). In the apical third, compared to coronal and middle third, the canal walls were often contaminated by inorganic debris and smear layer. Conclusion: All different concentrations of sodium hypochlorite activated with laser have a positive effect on removing of smear layer. Sodium hypochlorite activated with laser removed smear layer more effectively at the coronal and middle third compared to the apical third. PMID:28912942

Smear Layer Evaluation on Root Canal Preparation with Manual and Rotary Techniques using EDTA as an Irrigant: A Scanning Electron Microscopy Study

PubMed Central

Manjunatha, M; Annapurna, Kini; Sudhakar, V; Sunil Kumar, VC; Hiremath, Vinay Kumar; Shah, Ankur

2013-01-01

Introduction: The aim of any root canal treatment is to achieve a canal free of micro organisms, residual pulp remnants, debris and smear layer for the long term success of the procedure. Manual and automated instrumentation techniques along with proper irrigation regime is used to arrive at the aforementioned goal. Many authors focused on the preparation capabilities of various manual and rotary instruments but very few investigators stressed on the actual cleaning abilities of these instruments. Aims and objectives: This study was undertaken to evaluate the cleaning efficiency of manual K flex files and rotary Pro File systems in the root canals using a scanning electron microscope. Material and Methods:Thirty single rooted mandibular first premolars were divided into two groups and randomized (the manual group-M and the ProFile group-P) with respect to the preparation technique. The Manual group was hand instrumented with stainless steel K- Flexofiles by means of a conventional filing technique. The Pro File group was instrumented according to the manufacturer's instructions using a rotary handpiece. All canals were shaped and cleaned under frequent irrigation with EDTA. Final irrigation was carried out with 3 mL of normal saline solution to neutralize the action of the irrigant. The roots were split, one half of each tooth was selected for further SEM technique analysis and examined under the scanning electron microscope. The canal walls were quantitatively evaluated for the amount of debris and smear layer. The apical, middle and coronal regions of the canal surface, were graded (1-5) for debris and smear layer. A statistical analysis was performed using a Mann-Whitney Rank Sum test. ProFile performed least effective cleaning. Manual K-Flexofiles led to a grooved pattern. Results and Conclusion: A statistically significant difference was observed (p<0.05) between the two instrumentation techniques concerning the amount of debris and smear layer at the apical level. The manually filed canals had less debris and smear layer than those using a rotary technique. It was concluded from this study that none of the instrumentation techniques employed, produced the canal walls which were free of surface debris and smear layer. The manual instrumentation technique was better in cleaning the canals compared to the ProFile rotary Ni-Ti instruments despite the step-back technique used for manual instrumentation. How to cite this article: Manjunatha M, Kini A, Sudhakar V, Sunil K V C, Hiremath V K, Shah A. Smear Layer Evaluation on Root Canal Preparation with Manual and Rotary Techniques using EDTA as an Irrigant: A Scanning Electron Microscopy Study. J Int Oral Health 2013; 5(1):66-78. PMID:24155580

A Field-Tailored Reverse Transcription Loop-Mediated Isothermal Assay for High Sensitivity Detection of Plasmodium falciparum Infections

PubMed Central

Kemleu, Sylvie; Guelig, Dylan; Eboumbou Moukoko, Carole; Essangui, Estelle; Diesburg, Steven; Mouliom, Abas; Melingui, Bernard; Manga, Jeanne; Donkeu, Christiane; Epote, Annie; Texier, Gaëtan; LaBarre, Paul; Burton, Robert

2016-01-01

Highly sensitive and field deployable molecular diagnostic tools are critically needed for detecting submicroscopic, yet transmissible levels of malaria parasites prevalent in malaria endemic countries worldwide. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed and evaluated in comparison with thick blood smear microscopy, an antigen-based rapid diagnostic test (RDT), and an in-house RT-PCR targeting the same RT-LAMP transcript. The optimized assay detected Plasmodium falciparum infections in as little as 0.25ng of total parasite RNA, and exhibited a detection limit of 0.08 parasites/ μL when tested directly on infected whole blood lysates, or ~0.0008 parasites/ μL when using RNA extracts. Assay positivity was observed as early as eight minutes from initiation of the RT-LAMP and in most cases the reaction was complete before twenty minutes. Clinical evaluation of the assay on 132 suspected malaria cases resulted in a positivity rate of 90% for RT-LAMP using extracted RNA, and 85% when using whole blood lysates. The positivity rates were 70% for P. falciparum-specific RDT, 83% for RT-PCR, and 74% for thick blood smear microscopy (Mean parasite density = 36,986 parasites/ μL). Concordance rates between the developed RT-LAMP and comparator tests were greater than 75%, the lowest being with light microscopy (78%, McNemar’s test: P = 0.0002), and the highest was with RT-PCR (87%, McNemar’s test: P = 0.0523). Compared to reference RT-PCR, assay sensitivity was 90% for RT-LAMP on whole blood, and 96% for RT-LAMP using corresponding RNA extracts. Electricity-free heaters were further developed and evaluated in comparison with a battery-operated isothermal amplification machine for use with the developed test in resource-limited settings. Taken together, the data highlight the benefits of targeting high abundant RNA transcripts in molecular diagnosis, as well as the potential usefulness of the developed RT-LAMP-assay in malaria diagnosis in low to high parasite density settings. PMID:27824866

Failure of imidocarb dipropionate to eliminate Hepatozoon canis in naturally infected dogs based on parasitological and molecular evaluation methods.

PubMed

Sasanelli, Mariateresa; Paradies, Paola; Greco, Beatrice; Eyal, Osnat; Zaza, Valeria; Baneth, Gad

2010-08-04

The efficacy of imidocarb dipropionate for the treatment of Hepatozoon canis infection was studied in three naturally infected asymptomatic dogs followed longitudinally over 8 months. Response to treatment was followed by monitoring blood counts, parasitemia levels in blood, parasite in concentrated buffy-coat smears and by PCR. The dogs were initially treated with a low dose of 3 mg/kg imidocarb dipropionate twice a month and when parasitemia persisted after five treatments, with the regular dose of 6 mg/kg. In one dog, H. canis gamonts were no longer detectable by blood and buffy-coat microscopy after 2 months of therapy with 6 mg/kg while in the two other dogs gamonts were intermittently found in blood but persistently detectable in buffy-coat smears during the whole study period. Furthermore, combined therapy with doxycycline monohydrate administered at 10 mg/kg/day PO for 4 weeks also failed to eliminate H. canis. PCR revealed that parasite DNA was present in the blood of all dogs at all sampling dates regardless of treatment refuting the effectiveness of treatment suggested by negative blood microscopy. Detection of H. canis in buffy coat was found to be twice as sensitive than by blood smear and detection by PCR was even more sensitive revealing infection in eight samples (16% of total samples) negative by blood and buffy-coat microscopy. In conclusion, imidocarb dipropionate was not effective in eliminating H. canis from dogs treated repeatedly over 8 months. Microscopical detection is not sufficient for the evaluation of treatment response in H. canis infection and follow up by molecular techniques is recommended. Copyright 2010 Elsevier B.V. All rights reserved.

Flagging threshold optimization for manual blood smear review in primary care laboratory.

PubMed

Bihl, Pierre-Adrien

2018-04-01

Manual blood smear review is required when an anomaly detected by the automated hematologic analyzer triggers a flag. Our will through this study is to optimize these flagging thresholds for manual slide review in order to limit workload, while insuring clinical care through no extra false-negative. Flagging causes of 4,373 samples were investigated by manual slide review, after having been run on ADVIA 2120i. A set of 6 user-adjustments is proposed. By implementing all recommendations that we made, false-positive rate falls from 81.8% to 58.6%, while PPV increases from 18.2% to 23.7%. Hence, use of such optimized thresholds enables us to maximize efficiency without altering clinical care, but each laboratory should establish its own criteria to take into consideration local distinctive features.

Scanning electron microscopy comparison of the cleaning efficacy of a root canal system by Nd:YAG laser and rotary instruments.

PubMed

Samiei, Mohammad; Pakdel, Seyyed Mahdi Vahid; Rikhtegaran, Sahand; Shakoei, Sahar; Ebrahimpour, Delaram; Taghavi, Pedram

2014-08-01

This study evaluated the cleaning efficacy of a root canal system by Nd:YAG laser and rotary instruments. Sixty single-rooted human teeth were divided into four experimental groups (n=15). In the first group the teeth were prepared with a step-back technique using conventional K-files. In the second and third groups, tooth preparation was carried out using Nd:YAG laser and rotary NiTi instruments, respectively. Teeth in the fourth group were prepared by combined laser and rotary methods. The smear layer remaining on canal walls was then assessed by scanning electron microscopy in the coronal, middle, and apical portions. The comparison of smear layer removal efficacy between groups was carried out by Kruskal-Wallis and Mann-Whitney U tests. The mean grades of smear layer removal in rotary-laser, rotary, laser and step-back techniques were 1.34 ± 0.18, 2.2 ± 0.28, 1.91 ± 0.25, and 2.42 ± 0.19, respectively. On the whole, differences between rotary-laser and rotary groups, step-back, and the three other techniques (rotary, laser, and rotary-laser) were significant at p=0.034. Based on the findings of this study, the cleaning efficacy of rotary, laser, and rotary-laser techniques were better than the step-back technique and the combined laser and rotary technique was the most efficient method.

Comparative performance of PCR-based assay versus microscopy and culture for the direct detection of Mycobacterium tuberculosis in clinical respiratory specimens in Lebanon.

PubMed

Araj, G F; Talhouk, R S; Itani, L Y; Jaber, W; Jamaleddine, G W

2000-09-01

American University of Beirut Medical Center, Lebanon. To assess the performance of a polymerase chain reaction (PCR) using primers that flank 542 bp within IS6110 in Mycobacterium tuberculosis (TB) vs. microscopy and BACTEC culture, in the diagnosis of tuberculosis. A total of 82 clinical respiratory pulmonary specimens and 73 samples from BACTEC vials were tested by the three methods. Of 24 smear-positive culture-positive (SP-CP) and 11 smear-negative culture-positive (SN-CP) TB specimens, PCR detected 83% and 64%, respectively. Among 17 specimens yielding mycobacteria other than tuberculosis (MOTT), the PCR was positive in 33% SP-CP and 14% SN-CP specimens. Among the 73 BACTEC vials, PCR was positive in 36 of 38 (95%) yielding culture-positive TB, and in one of 20 (5%) yielding culture positive MOTT. None of the 30 smear-negative culture-negative (SN-CN) clinical specimens and 15 of the CN vials were positive by PCR. The overall sensitivity of PCR was 77% and 95% for TB detection in respiratory specimens and BACTEC vials, respectively, and the specificity was 94% in both. Because a substantial number of TB cases are missed, especially in SN-CP specimens, a PCR-based assay utilizing these primers cannot be used reliably, alone, in clinical laboratory diagnosis of mycobacterial respiratory infections.

Cleaning capacity of octenidine as root canal irrigant: A scanning electron microscopy study.

PubMed

Coaguila-Llerena, Hernán; Stefanini da Silva, Virgínia; Tanomaru-Filho, Mario; Guerreiro Tanomaru, Juliane Maria; Faria, Gisele

2018-06-01

The aim of this study was to assess the cleaning capacity of the octenidine hydrochloride (OCT) used as root canal irrigant by scanning electron microscopy (SEM) analysis. Sixty human unirradicular extracted teeth were randomly distributed in 6 groups (n = 10) according to irrigant solutions which were used during root canal preparation: G1, 0.1% OCT; G2, 2% chlorhexidine (CHX); G3, 2.5% sodium hypochlorite (NaOCl); G4, OCT + 17% ethylenediaminetetraacetic acid (EDTA); G5, 2.5% NaOCl + 17% EDTA and G6, distilled water. All specimens were instrumented with ProTaper system up to F4. Teeth were sectioned and prepared for SEM. The smear layer was evaluated using a 5-score system and the data were analyzed by Kruskal-Wallis and Dunn (α = 0.05). In all root canal thirds there was no significant difference between OCT, CHX, NaOCl, and water groups (p > .05), and these groups showed higher smear layer values than NaOCl + EDTA and OCT + EDTA groups (p < .05). There was no significant difference between NaOCl + EDTA and OCT + EDTA groups (p > .05). It was concluded that OCT used as a single root canal irrigant presented poor cleaning capacity and could be used in association with a final irrigation with EDTA to obtain smear layer removal. © 2018 Wiley Periodicals, Inc.

Laboratory diagnosis of tuberculosis: Advances in technology and drug susceptibility testing.

PubMed

Oommen, Seema; Banaji, Nandita

2017-01-01

There have been rapid technological advances in the detection of Mycobacterium tuberculosis and its drug susceptibility in clinical samples. These include advances in microscopic examination, in vitro culture and application of molecular techniques. The World Health Organization (WHO) has played a large role in evaluating these technologies for their efficacy and feasibility, especially in the developing countries. Amongst these, the Revised National Tuberculosis Control Programme (RNTCP), through its national network of designated microscopy centres and intermediate reference laboratories, has adopted certain technologies that are currently implemented in India. Advances in microscopy technology include fluorescent microscopy using light-emitting diode source, sodium hypochlorite microscopy and vital fluorescent staining of sputum smears. Automation of in vitro culture has markedly reduced the turnaround time (TAT), even in smear-negative samples, and permits simultaneous detection of resistant mutants. Molecular detection of drug resistance has further reduced the TAT, and the cartridge-based nucleic acid amplification test with its performance convenience and rapid results, appears poised to become the future of tuberculosis (TB) diagnosis in all settings, provided the cost of testing is reduced especially for use in private diagnostic settings. This article reviews technologies currently available for the diagnosis of TB, keeping in mind the WHO recommendations and the RNTCP practices. This is a thematic synthesis of data available on diagnosis in literature, preserving the conclusions of the primary studies.

Bottom-up or top-down: unit cost estimation of tuberculosis diagnostic tests in India.

PubMed

Rupert, S; Vassall, A; Raizada, N; Khaparde, S D; Boehme, C; Salhotra, V S; Sachdeva, K S; Nair, S A; Hoog, A H Van't

2017-04-01

Of 18 sites that participated in an implementation study of the Xpert® MTB/RIF assay in India, we selected five microscopy centres and two reference laboratories. To obtain unit costs of diagnostic tests for tuberculosis (TB) and drug-resistant TB. Laboratories were purposely selected to capture regional variations and different laboratory types. Both bottom-up and the top-down methods were used to estimate unit costs. At the microscopy centres, mean bottom-up unit costs were respectively US$0.83 (range US$0.60-US$1.10) and US$12.29 (US$11.61-US$12.89) for sputum smear microscopy and Xpert. At the reference laboratories, mean unit costs were US$1.69 for the decontamination procedure, US$9.83 for a solid culture, US$11.06 for a liquid culture, US$29.88 for a drug susceptibility test, and US$18.18 for a line-probe assay. Top-down mean unit cost estimates were higher for all tests, and for sputum smear microscopy and Xpert these increased to respectively US$1.51 and US$13.58. The difference between bottom-up and top-down estimates was greatest for tests performed at the reference laboratories. These unit costs for TB diagnostics can be used to estimate resource requirements and cost-effectiveness in India, taking into account geographical location, laboratory type and capacity utilisation.

Detection of selected intestinal helminths and protozoa at Hospital Universiti Sains Malaysia using multiplex real-time PCR.

PubMed

Basuni, M; Mohamed, Z; Ahmad, M; Zakaria, N Z; Noordin, R

2012-09-01

Intestinal parasites are the causative agents of a number of important human infections in developing countries. The objective of this study was to determine the prevalence of selected helminths and protozoan infections among patients admitted with gastrointestinal disorders at Hospital Universiti Sains Malaysia, Kelantan, Malaysia using multiplex real-time PCR. In addition microscopic examination was also performed following direct smear, zinc sulphate concentration and Kato-Katz thick smear techniques; and the presence of protozoan parasites was confirmed using trichrome and acid-fast stains. Of the 225 faecal samples analysed, 26.2% were positive for intestinal parasites by the multiplex real-time PCR, while 5.3% were positive by microscopy. As compared to microscopy, the multiplex real-time PCR detected 5.8 and 4.5 times more positives for the selected helminth and protozoan infections respectively. Among the selected helminths detected in this study, hookworm was the most prevalent by real-time PCR, while Ascaris lumbricoides was detected the most by microscopy. Meanwhile, among the selected protozoa detected in this study, Entamoeba histolytica was the most prevalent by real-time PCR, however microscopy detected equal number of cases with E. histolytica and Giardia lamblia. This study showed that real-time PCR can be used to obtain a more accurate prevalence data on intestinal helminths and protozoa.

Staining-free malaria diagnostics by multispectral and multimodality light-emitting-diode microscopy

NASA Astrophysics Data System (ADS)

Merdasa, Aboma; Brydegaard, Mikkel; Svanberg, Sune; Zoueu, Jeremie T.

2013-03-01

We report an accurate optical differentiation technique between healthy and malaria-infected erythrocytes by quasi-simultaneous measurements of transmittance, reflectance, and scattering properties of unstained blood smears using a multispectral and multimode light-emitting diode microscope. We propose a technique for automated imaging, identification, and counting of malaria-infected erythrocytes for real-time and cost-effective parasitaemia diagnosis as an effective alternative to the manual screening of stained blood smears, now considered to be the gold standard in malaria diagnosis. We evaluate the performance of our algorithm against manual estimations of an expert and show a spectrally resolved increased scattering from malaria-infected blood cells.

Multidrug-resistant tuberculosis treatment failure detection depends on monitoring interval and microbiological method

PubMed Central

White, Richard A.; Lu, Chunling; Rodriguez, Carly A.; Bayona, Jaime; Becerra, Mercedes C.; Burgos, Marcos; Centis, Rosella; Cohen, Theodore; Cox, Helen; D'Ambrosio, Lia; Danilovitz, Manfred; Falzon, Dennis; Gelmanova, Irina Y.; Gler, Maria T.; Grinsdale, Jennifer A.; Holtz, Timothy H.; Keshavjee, Salmaan; Leimane, Vaira; Menzies, Dick; Milstein, Meredith B.; Mishustin, Sergey P.; Pagano, Marcello; Quelapio, Maria I.; Shean, Karen; Shin, Sonya S.; Tolman, Arielle W.; van der Walt, Martha L.; Van Deun, Armand; Viiklepp, Piret

2016-01-01

Debate persists about monitoring method (culture or smear) and interval (monthly or less frequently) during treatment for multidrug-resistant tuberculosis (MDR-TB). We analysed existing data and estimated the effect of monitoring strategies on timing of failure detection. We identified studies reporting microbiological response to MDR-TB treatment and solicited individual patient data from authors. Frailty survival models were used to estimate pooled relative risk of failure detection in the last 12 months of treatment; hazard of failure using monthly culture was the reference. Data were obtained for 5410 patients across 12 observational studies. During the last 12 months of treatment, failure detection occurred in a median of 3 months by monthly culture; failure detection was delayed by 2, 7, and 9 months relying on bimonthly culture, monthly smear and bimonthly smear, respectively. Risk (95% CI) of failure detection delay resulting from monthly smear relative to culture is 0.38 (0.34–0.42) for all patients and 0.33 (0.25–0.42) for HIV-co-infected patients. Failure detection is delayed by reducing the sensitivity and frequency of the monitoring method. Monthly monitoring of sputum cultures from patients receiving MDR-TB treatment is recommended. Expanded laboratory capacity is needed for high-quality culture, and for smear microscopy and rapid molecular tests. PMID:27587552

Effectiveness of Three Different Irrigants - 17% Ethylenediaminetetraacetic Acid, Q-MIX, and Phytic Acid in Smear Layer Removal: A Comparative Scanning Electron Microscope Study.

PubMed

Jagzap, Janhavi Balasaheb; Patil, Sanjay S; Gade, Vandana Jaykumar; Chandhok, Deepika J; Upagade, Madhura A; Thakur, Deepa A

2017-01-01

Removal of smear layer from the root canal walls is important for long-standing endodontic success. The aim of this study is to evaluate and compare smear layer removing ability among 17% ethylenediaminetetraacetic acid (EDTA), Q-MIX, and phytic acid by scanning electron microscopy (SEM). This in-vitro experimental study assessed smear layer removal using three different irrigants. Thirty single-rooted freshly extracted human permanent premolars were collected, disinfected, and decoronated to a standardized root length of 13 mm. Root canals were cleaned and shaped till F2 universal rotary protaper at working length 1 mm short of the apex. They were randomly divided into three groups, and final irrigation was done accordingly. Group 1 ( n = 10): with 1 ml of 17% EDTA, Group 2 ( n = 10): with 1 ml of Q-MIX, Group 3 ( n = 10): with 1 ml of phytic acid. Samples were then longitudinally sectioned and evaluated under SEM at coronal, middle, and apical levels. Two-way analysis of variance and Tukey's post hoc test were performed. The level of significance was set at 0.05. Smear layer removing ability among irrigants and sections in descending order: 17 EDTA > Q-MIX > phytic acid; coronal > middle > apical. 17% EDTA showed better and promising results followed by Q-MIX and then phytic acid.

Effects of ethylenediaminetetraacetic, etidronic and peracetic acid irrigation on human root dentine and the smear layer.

PubMed

Lottanti, S; Gautschi, H; Sener, B; Zehnder, M

2009-04-01

To evaluate the effects of ethylenediaminetetraacetic (EDTA), etidronic (EA) and peracetic acid (PA) when used in conjunction with sodium hypochlorite (NaOCl) as root canal irrigants on calcium eluted from canals, smear layer, and root dentine demineralization after instrumentation/irrigation. Single-rooted human premolars were irrigated as follows (n = 12 per group): (1) 1% NaOCl during instrumentation, deionized water after instrumentation, (2) 1% NaOCl during, 17% EDTA after instrumentation, (3) a 1 : 1-mixture of 2% NaOCl and 18% EA during and after instrumentation, and (4) 1% NaOCl during, 2.25% PA after instrumentation. Irrigant volumes and contact times were 10 mL/15 min during and 5 mL/3 min after instrumentation. The evaluated outcomes were eluted calcium by atomic absorption spectroscopy, smear-covered areas by scanning electron microscopy in secondary electron mode and apparent canal wall decalcifications on root transsections in backscatter mode. For the smear layer analysis, sclerotic dentine was taken into consideration. Results were compared using appropriate parametric and nonparametric tests, alpha = 0.05. The statistical comparison of the protocols regarding calcium elution revealed that protocol (1) yielded less calcium than (3), which yielded less than protocols (2) and (4). Most of the instrumented canal walls treated with one of the decalcifying agents were free of smear layer. Protocols (1) and (3) caused no decalcification of root dentine, whilst (2) and (4) showed substance typical demineralization patterns. The decalcifying agents under investigation were all able to remove or prevent a smear layer. However, they eroded the dentine wall differently.

Comparative Study of GeneXpert with ZN Stain and Culture in Samples of Suspected Pulmonary Tuberculosis

PubMed Central

Bajaj, Ashish; Bhatia, Vinay; Dutt, Sarjana

2016-01-01

Introduction Tuberculosis remains one of the deadliest communicable diseases. There are number of tests available for the diagnosis of tuberculosis but conventional microscopy has low sensitivity and culture although gold standard, but takes longer time for positivity. On the other side, Nucleic acid amplification techniques due to its rapidity and sensitivity not only help in early diagnosis and management of tuberculosis especially in patients with high clinical suspicion like immunocompromised patients, history of contact with active tuberculosis patient etc., but also curtail the transmission of the disease. Aim To evaluate the sensitivity, specificity, positive predictive value and negative predictive value of Nucleic acid amplification assay (GeneXpert) using respiratory samples in patients with suspected pulmonary tuberculosis and compare with AFB smear microscopy (Ziehl Neelsen stain) and Acid Fast Bacilli (AFB) culture. Materials and Methods We retrospectively reviewed the respiratory samples of suspected pulmonary tuberculosis (including Bronchoalveolar lavage and sputum) of 170 patients from Jan 2015 to Nov 2015 for ZN stain, culture and GeneXpert (Xpert® MTB/Rif assay). The sensitivity, specificity, PPV and NPV of GeneXpert and ZN microscopy were calculated using Liquid culture of Mycobacterium tuberculosis as gold standard. Results A total of 170 patient samples were evaluated in final analysis. Of these, 14 samples were positive by all three methods used in our study. The overall sensitivity, specificity, PPV and NPV of GeneXpert were 86.8%, 93.1%, 78.5% and 96% respectively and for BAL sample, 81.4%, 93.4%, 73.3% and 95.7% respectively. The overall sensitivity and specificity of AFB smear microscopy were 22.2%, % and 78.5% respectively and for BAL sample 22.2% and 100% respectively. For AFB negative samples sensitivity and specificity were 79.1% and 93.1% respectively. Conclusion GeneXpert has a higher sensitivity than AFB smear microscopy in respiratory samples. GeneXpert can be a useful tool for early diagnosis of patients with high clinical suspicion of pulmonary tuberculosis. Positive GeneXpert, but culture negative results should be read cautiously and be well correlated with clinical and treatment history of the patient. The other major advantage of Gene Xpert is that it simultaneously detects Rifampicin resistance and especially beneficial in patient with MDR and HIV associated tuberculosis and should be studied further. PMID:27437212

Cost-effectiveness of diagnostic for malaria in Extra-Amazon Region, Brazil

PubMed Central

2012-01-01

Background Rapid diagnostic tests (RDT) for malaria have been demonstrated to be effective and they should replace microscopy in certain areas. Method The cost-effectiveness of five RDT and thick smear microscopy was estimated and compared. Data were collected on Brazilian Extra-Amazon Region. Data sources included the National Malaria Control Programme of the Ministry of Health, the National Healthcare System reimbursement table, laboratory suppliers and scientific literature. The perspective was that of the Brazilian public health system, the analytical horizon was from the start of fever until the diagnostic results provided to patient and the temporal reference was that of year 2010. Two costing methods were produced, based on exclusive-use microscopy or shared-use microscopy. The results were expressed in costs per adequately diagnosed cases in 2010 U.S. dollars. One-way sensitivity analysis was performed considering key model parameters. Results In the cost-effectiveness analysis with exclusive-use microscopy, the RDT CareStart™ was the most cost-effective diagnostic strategy. Microscopy was the most expensive and most effective, with an additional case adequately diagnosed by microscopy costing US$ 35,550.00 in relation to CareStart™. In opposite, in the cost-effectiveness analysis with shared-use microscopy, the thick smear was extremely cost-effective. Introducing into the analytic model with shared-use microscopy a probability for individual access to the diagnosis, assuming a probability of 100% of access for a public health system user to any RDT and, hypothetically, of 85% of access to microscopy, this test saw its effectiveness reduced and was dominated by the RDT CareStart™. Conclusion The analysis of cost-effectiveness of malaria diagnosis technologies in the Brazilian Extra-Amazon Region depends on the exclusive or shared use of the microscopy. Following the assumptions of this study, shared-use microscopy would be the most cost-effective strategy of the six technologies evaluated. However, if used exclusively for diagnosing malaria, microscopy would be the worst use of resources. Microscopy would not be the most cost-effective strategy, even when structure is shared with other programmes, when the probability of a patient having access to it was reduced. Under these circumstances, the RDT CareStart™ would be the most cost-effective strategy. PMID:23176717

High resolution computational on-chip imaging of biological samples using sparsity constraint (Conference Presentation)

NASA Astrophysics Data System (ADS)

Rivenson, Yair; Wu, Chris; Wang, Hongda; Zhang, Yibo; Ozcan, Aydogan

2017-03-01

Microscopic imaging of biological samples such as pathology slides is one of the standard diagnostic methods for screening various diseases, including cancer. These biological samples are usually imaged using traditional optical microscopy tools; however, the high cost, bulkiness and limited imaging throughput of traditional microscopes partially restrict their deployment in resource-limited settings. In order to mitigate this, we previously demonstrated a cost-effective and compact lens-less on-chip microscopy platform with a wide field-of-view of >20-30 mm^2. The lens-less microscopy platform has shown its effectiveness for imaging of highly connected biological samples, such as pathology slides of various tissue samples and smears, among others. This computational holographic microscope requires a set of super-resolved holograms acquired at multiple sample-to-sensor distances, which are used as input to an iterative phase recovery algorithm and holographic reconstruction process, yielding high-resolution images of the samples in phase and amplitude channels. Here we demonstrate that in order to reconstruct clinically relevant images with high resolution and image contrast, we require less than 50% of the previously reported nominal number of holograms acquired at different sample-to-sensor distances. This is achieved by incorporating a loose sparsity constraint as part of the iterative holographic object reconstruction. We demonstrate the success of this sparsity-based computational lens-less microscopy platform by imaging pathology slides of breast cancer tissue and Papanicolaou (Pap) smears.

Hydrophilic-treated plastic plates for wide-range analysis of Giemsa-stained red blood cells and automated Plasmodium infection rate counting.

PubMed

Hashimoto, Muneaki; Yatsushiro, Shouki; Yamamura, Shohei; Tanaka, Masato; Sakamoto, Hirokazu; Ido, Yusuke; Kajimoto, Kazuaki; Bando, Mika; Kido, Jun-Ichi; Kataoka, Masatoshi

2017-08-08

Malaria is a red blood cell (RBC) infection caused by Plasmodium parasites. To determine RBC infection rate, which is essential for malaria study and diagnosis, microscopic evaluation of Giemsa-stained thin blood smears on glass slides ('Giemsa microscopy') has been performed as the accepted gold standard for over 100 years. However, only a small area of the blood smear provides a monolayer of RBCs suitable for determination of infection rate, which is one of the major reasons for the low parasite detection rate by Giemsa microscopy. In addition, because Giemsa microscopy is exacting and time-consuming, automated counting of infection rates is highly desirable. A method that allows for microscopic examination of Giemsa-stained cells spread in a monolayer on almost the whole surface of hydrophilic-treated cyclic olefin copolymer (COC) plates was established. Because wide-range Giemsa microscopy can be performed on a hydrophilic-treated plate, the method may enable more reliable diagnosis of malaria in patients with low parasitaemia burden. Furthermore, the number of RBCs and parasites stained with a fluorescent nuclear staining dye could be counted automatically with a software tool, without Giemsa staining. As a result, researchers studying malaria may calculate the infection rate easily, rapidly, and accurately even in low parasitaemia. Because the running cost of these methods is very low and they do not involve complicated techniques, the use of hydrophilic COC plates may contribute to improved and more accurate diagnosis and research of malaria.

Comparison of PCR-Based Diagnosis with Centrifuged-Based Enrichment Method for Detection of Borrelia persica in Animal Blood Samples.

PubMed

Naddaf, S R; Kishdehi, M; Siavashi, Mr

2011-01-01

The mainstay of diagnosis of relapsing fever (RF) is demonstration of the spirochetes in Giemsa-stained thick blood smears, but during non fever periods the bacteria are very scanty and rarely detected in blood smears by microscopy. This study is aimed to evaluate the sensitivity of different methods developed for detection of low-grade spirochetemia. Animal blood samples with low degrees of spirochetemia were tested with two PCRs and a nested PCR targeting flaB, GlpQ, and rrs genes. Also, a centrifuged-based enrichment method and Giemsa staining were performed on blood samples with various degrees of spirochetemia. The flaB-PCR and nested rrs-PCR turned positive with various degrees of spirochetemia including the blood samples that turned negative with dark-field microscopy. The GlpQ-PCR was positive as far as at least one spirochete was seen in 5-10 microscopic fields. The sensitivity of GlpQ-PCR increased when DNA from Buffy Coat Layer (BCL) was used as template. The centrifuged-based enrichment method turned positive with as low concentration as 50 bacteria/ml blood, while Giemsa thick staining detected bacteria with concentrations ≥ 25000 bacteria/ml. Centrifuged-based enrichment method appeared as much as 500-fold more sensitive than thick smears, which makes it even superior to some PCR assays. Due to simplicity and minimal laboratory requirements, this method can be considered a valuable tool for diagnosis of RF in rural health centers.

Effects of NiTi rotary and reciprocating instruments on debris and smear layer scores: an SEM evaluation.

PubMed

Poggio, Claudio; Dagna, Alberto; Chiesa, Marco; Scribante, Andrea; Beltrami, Riccardo; Colombo, Marco

2014-12-30

The purpose of the present study was to investigate the cleaning efficacy of 2 different nickel-titanium (NiTi) rotary systems: a new single system Reciproc compared with the rotary full sequence Mtwo. The amount of debris and the morphology of smear layer were the parameters for the evaluation of the cleanliness of root canals. The null hypothesis of the study was that there would be no significant difference in debris scores and smear layer scores between the 2 systems. 
 Forty single-rooted freshly extracted teeth were selected and divided into 2 groups. For each group, all root canals were shaped with 2 different NiTi instruments (Mtwo and Reciproc) and irrigated with 5.25% NaOCl and 17% ethylenediaminetetraacetic acid (EDTA) solutions by 3 trained operators. Specimens were fractured longitudinally and prepared for scanning electron microscopy (SEM) analysis at a standard magnification of ×5,000. The presence/absence of smear layer and the presence/absence of debris at the coronal, middle and apical third of each canal were evaluated using two 5-step scales for scores. Numeric data were analyzed using the Kruskal-Wallis and Mann-Whitney U statistical tests, and significance was set at a P value <0.05. 
 This study revealed significant differences among the various groups. The Mtwo group presented significantly lower smear layer scores than the Reciproc group (P<0.01) in the middle and apical third of the canal. 
 Conventional continuous rotary NiTi instruments associated with 5.25% NaOCl and 17% EDTA solutions seem to be better for obtaining clean dentinal canal walls. Reciprocating instruments leave a higher quantity of smear layer which covers dentinal walls and dentinal tubules.

Audit of the practice of sputum smear examination for patients with suspected pulmonary tuberculosis in Fiji.

PubMed

Gounder, Shakti; Tayler-Smith, Katherine; Khogali, Mohammed; Raikabula, Maopa; Harries, Anthony D

2013-07-01

In Fiji, patients with suspected pulmonary tuberculosis (PTB) currently submit three sputum specimens for smear microscopy for acid-fast bacilli, but there is little information about how well this practice is carried out. A cross-sectional retrospective review was carried out in all four TB diagnostic laboratories in Fiji to determine among new patients presenting with suspected PTB in 2011: the quality of submitted sputum; the number of sputum samples submitted; the relationship between quality and number of submitted samples to smear-positivity; and positive yield from first, second and third samples. Of 1940 patients with suspected PTB, 3522 sputum samples were submitted: 997 (51.4%) patients submitted one sample, 304 (15.7%) patients submitted two samples and 639 (32.9%) submitted three samples. Sputum quality was recorded in 2528 (71.8%) of samples, of which 1046 (41.4%) were of poor quality. Poor quality sputum was more frequent in females, inpatients and children (0-14 years). Good quality sputum and a higher number of submitted samples positively correlated with smear-positivity for acid-fast bacilli. There were 122 (6.3%) patients with suspected PTB who were sputum smear positive. Of those, 89 had submitted three sputum samples: 79 (89%) were diagnosed based on the first sputum sample, 6 (7%) on the second sample and 4 (4%) on the third sample. This study shows that there are deficiencies in the practice of sputum smear examination in Fiji with respect to sputum quality and recommended number of submitted samples, although the results support the continued use of three sputum samples for TB diagnosis. Ways to improve sputum quality and adherence to recommended guidelines are needed.

Morphologic characterization of specific granules in Greyhound eosinophils.

PubMed

Iazbik, M C; Couto, C G

2005-06-01

"Vacuolated" eosinophils (ie, eosinophils with empty, nonstaining granules) have been described previously in normal Greyhounds. However, to our knowledge, detailed studies of granules in vacuolated and normal eosinophils in this breed have not been performed. The objective of this prospective study was to characterize some of the morphologic, ultrastructural, and cytochemical staining features of specific (primary) granules in both normal and vacuolated eosinophils in Greyhound blood. Morphologic features of eosinophils in Wright's- and Diff-Quik-stained peripheral blood smears from 49 Greyhounds were compared with 200 blood smears from non-Greyhound dogs. Transmission electron microscopy was done on blood from 3 Greyhounds with vacuolated eosinophils and 3 with normal eosinophil granules. Blood smears from 4 of these dogs also were stained cytochemically with alkaline phosphatase (AP), chloracetate esterase (CAE), and alpha naphthyl butyrate esterase (ANBE). The morphologic features and tinctorial properties of vacuolated and normal eosinophils were compared. Twenty-six Greyhounds (53%) had vacuolated eosinophils and 23 (47%) had normal granulated eosinophils in smears stained with Wright's stain. Only 1% of eosinophils were vacuolated in non-Greyhound dogs. Twenty of the 23 (85%) Greyhounds with normal granulated eosinophils on Wright's-stained smears had vacuolated eosinophils in smears stained with Diff-Quik. Ultrastructurally, no morphologic differences were observed between granules of vacuolated and normal eosinophils. Both vacuolated and normal eosinophils in Greyhounds were positive for AP and negative for CAE and ANBE, as expected for normal dogs. Vacuolated eosinophils in Greyhounds likely reflect, at least in part, differential staining properties of the specific granules with different hematologic stains. Ultrastuctural and cytochemical features of eosinophil granules were similar in normal and vacuolated eosinophils from Greyhounds.

The self-adjusting file (SAF). Part 3: removal of debris and smear layer-A scanning electron microscope study.

PubMed

Metzger, Zvi; Teperovich, Ehud; Cohen, Raphaela; Zary, Raviv; Paqué, Frank; Hülsmann, Michael

2010-04-01

The aim of this study was to evaluate the cleaning ability of the Self-Adjusting File (SAF) system in terms of removal of debris and smear layer. Root canal preparations were performed in 20 root canals using an SAF operated with a continuous irrigation device. The glide path was initially established using a size 20 K-file followed by the SAF file that was operated in the root canal via a vibrating motion for a total of 4 minutes. Sodium hypochlorite (3%) and EDTA (17%) were used as continuous irrigants and were alternated every minute during this initial 4-minute period. This was followed by a 30-second rinse using EDTA applied through a nonactivated SAF and a final flush with sodium hypochlorite. The roots were split longitudinally and subjected to scanning electron microscopy (SEM). The presence of debris and a smear layer in the coronal, middle, and apical thirds of the canal were evaluated through the analysis of the SEM images using five-score evaluation systems based on reference photographs. The SAF operation with continuous irrigation, using alternating irrigants, resulted in root canal walls that were free of debris in all thirds of the canal in all (100%) of the samples. In addition, smear layer-free surfaces were observed in 100% and 80% of the coronal and middle thirds of the canal, respectively. In the apical third of the canal, smear layer-free surfaces were found in 65% of the root canals. The operation of the SAF system with continuous irrigation coupled with alternating sodium hypochlorite and EDTA treatment resulted in a clean and mostly smear layer-free dentinal surface in all parts of the root canal. Copyright (c) 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Use of exfoliative cytology for diagnosis of transmissible venereal tumour and controlling the recovery period in the bitch.

PubMed

Erünal-Maral, N; Findik, M; Aslan, S

2000-05-01

Transmissible venereal tumour (TVT) is a rare vaginal tumour that can be treated surgically or cryosurgically as well as by radiotherapy or chemotherapy. Vincristine has been found to be very effective for treating TVT. Since vaginal secretion or discharge may contain neoplastic cells, the cytological identification of TVT cells is possible. The present study was carried out in 12 bitches. Vaginal smears were obtained with cotton swab from the anterior vagina and TVT suspected structures. The smears were stained according to Papanicolaou and assessed by light microscopy. Additionally the general condition of the patients was evaluated by haematological and radiographic examinations. In bitches with TVT vincristine sulphate was administered intravenously at weekly intervals. The total treatment period was three to six weeks until no atypical cells were found in the smear. This was the case after an average of 3.2 +/- 1.3 applications. Tumour masses became smaller and by this non-visible from the rima vulva after 4.2 +/- 0.7 applications. During the treatment, two of the 12 bitches (16.7%) suffered from vomiting and diarrhoea while three (25%) showed neutropenia. Twelve months after completion of treatment, the bitches were examined again and vaginal smears were taken in order to control the recovery process or a possible recurrence of TVT. No atypical cells were found in any vaginal smear. By this exfoliative cytology has proved to be a safe and easy method for TVT diagnosis and for observing the recovery process.

A novel sputum transport solution eliminates cold chain and supports routine tuberculosis testing in Nepal.

PubMed

Maharjan, Bhagwan; Shrestha, Bhabana; Weirich, Alexandra; Stewart, Andrew; Kelly-Cirino, Cassandra D

2016-12-01

This preliminary study evaluated the transport reagent OMNIgene SPUTUM (OMS) in a real-world, resource-limited setting: a zonal hospital and national tuberculosis (TB) reference laboratory, Nepal. The objectives were to: (1) assess the performance of OMS for transporting sputum from peripheral sites without cold chain stabilization; and (2) compare with Nepal's standard of care (SOC) for Mycobacterium tuberculosis smear and culture diagnostics. Sixty sputa were manually split into a SOC sample (airline-couriered to the laboratory, conventional processing) and an OMS sample (OMS added at collection, no cold chain transport or processing). Smear microscopy and solid culture were performed. Transport was 0-8days. Forty-one samples (68%) were smear-positive using both methods. Of the OMS cultures, 37 (62%) were positive, 22 (36%) were negative, and one (2%) was contaminated. Corresponding SOC results were 32 (53%), 21 (35%), and seven (12%). OMS "rescued" six (i.e., missed using SOC) compared with one rescue using SOC. Of smear-positives, six SOC samples produced contaminated cultures whereas only one OMS sample was contaminated. OMS reduced culture contamination from 12% to 2%, and improved TB detection by 9%. The results suggest that OMS could perform well as a no cold chain, long-term transport solution for smear and culture testing. The findings provide a basis for larger feasibility studies. Copyright © 2016 Ministry of Health, Saudi Arabia. Published by Elsevier Ltd. All rights reserved.

Cost-effectiveness analysis of the Xpert MTB/RIF assay for rapid diagnosis of suspected tuberculosis in an intermediate burden area.

PubMed

You, Joyce H S; Lui, Grace; Kam, Kai Man; Lee, Nelson L S

2015-04-01

We examined, from a Hong Kong healthcare providers' perspective, the cost-effectiveness of rapid diagnosis with Xpert in patients hospitalized for suspected active pulmonary tuberculosis (PTB). A decision tree was designed to simulate outcomes of three diagnostic assessment strategies in adult patients hospitalized for suspected active PTB: conventional approach, sputum smear plus Xpert for acid-fast bacilli (AFB) smear-negative, and a single sputum Xpert test. Model inputs were derived from the literature. Outcome measures were direct medical cost, one-year mortality rate, quality-adjusted life-years (QALYs) and incremental cost per QALY (ICER). In the base-case analysis, Xpert was more effective with higher QALYs gained and a lower mortality rate when compared with smear plus Xpert by an ICER of USD99. A conventional diagnostic approach was the least preferred option with the highest cost, lowest QALYs gained and highest mortality rate. Sensitivity analysis showed that Xpert would be the most cost-effective option if the sensitivity of sputum AFB smear microscopy was ≤74%. The probabilities of Xpert, smear plus Xpert and a conventional approach to be cost-effective were 94.5%, 5.5% and 0%, respectively, in 10,000 Monte Carlo simulations. The Xpert sputum test appears to be a highly cost-effective diagnostic strategy for patients with suspected active PTB in an intermediate burden area like Hong Kong. Copyright © 2015 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

Fluorescence In Situ Hybridization with Peptide Nucleic Acid Probes for Rapid Identification of Candida albicans Directly from Blood Culture Bottles

PubMed Central

Rigby, Susan; Procop, Gary W.; Haase, Gerhard; Wilson, Deborah; Hall, Geraldine; Kurtzman, Cletus; Oliveira, Kenneth; Von Oy, Sabina; Hyldig-Nielsen, Jens J.; Coull, James; Stender, Henrik

2002-01-01

A new fluorescence in situ hybridization (FISH) method that uses peptide nucleic acid (PNA) probes for identification of Candida albicans directly from positive-blood-culture bottles in which yeast was observed by Gram staining (herein referred to as yeast-positive blood culture bottles) is described. The test (the C. albicans PNA FISH method) is based on a fluorescein-labeled PNA probe that targets C. albicans 26S rRNA. The PNA probe is added to smears made directly from the contents of the blood culture bottle and hybridized for 90 min at 55°C. Unhybridized PNA probe is removed by washing of the mixture (30 min), and the smears are examined by fluorescence microscopy. The specificity of the method was confirmed with 23 reference strains representing phylogenetically related yeast species and 148 clinical isolates covering the clinically most significant yeast species, including C. albicans (n = 72), C. dubliniensis (n = 58), C. glabrata (n = 5), C. krusei (n = 2), C. parapsilosis (n = 4), and C. tropicalis (n = 3). The performance of the C. albicans PNA FISH method as a diagnostic test was evaluated with 33 routine and 25 simulated yeast-positive blood culture bottles and showed 100% sensitivity and 100% specificity. It is concluded that this 2.5-h method for the definitive identification of C. albicans directly from yeast-positive blood culture bottles provides important information for optimal antifungal therapy and patient management. PMID:12037084

Operational implementation of LED fluorescence microscopy in screening tuberculosis suspects in an urban HIV clinic in Uganda.

PubMed

Albert, Heidi; Nakiyingi, Lydia; Sempa, Joseph; Mbabazi, Olive; Mukkada, Sheena; Nyesiga, Barnabas; Perkins, Mark D; Manabe, Yukari C

2013-01-01

Light emitting diode (LED) fluorescence microscopy (FM) is an affordable, technology targeted for use in resource-limited settings and recommended for widespread roll-out by the World Health Organization (WHO). We sought to compare the operational performance of three LED FM methods compared to light microscopy in a cohort of HIV-positive tuberculosis (TB) suspects at an urban clinic in a high TB burden country. Two spot specimens collected from TB suspects were included in the study. Smears were stained using auramine O method and read after blinding by three LED-based FM methods by trained laboratory technicians in the Infectious Diseases Institutelaboratory. Leftover portions of the refrigerated sputum specimens were transported to the FIND Tuberculosis Research Laboratory for Ziehl Neelsen (ZN) smear preparation and reading by experienced technologist as well as liquid and solid culture. 174 of 627 (27.8%) specimens collected yielded one or more positive mycobacterial cultures. 94.3% (164/174) were M. tuberculosis complex. LED FM was between 7.3-11.0% more sensitive compared to ZN microscopy. Of the 592 specimens examined by all microscopy methods, there was no significant difference in sensitivity between the three LED FM methods. The specificity of the LED FM methods was between 6.1% and 7.7% lower than ZN microscopy (P<0.001), although exclusion of the single poor reader resulted in over 98% specificity for all FM methods. Laboratory technicians in routine settings can be trained to use FM which is more sensitive than ZN microscopy. Despite rigorous proficiency testing, there were operator-dependent accuracy issues which highlight the critical need for intensive quality assurance procedures during LED FM implementation. The low sensitivity of FM for HIV-positive individuals particularly those with low CD4 T cell counts, will limit the number of additional patients found by LED FM in countries with high rates of HIV co-infection.

Operational Implementation of LED Fluorescence Microscopy in Screening Tuberculosis Suspects in an Urban HIV Clinic in Uganda

PubMed Central

Albert, Heidi; Nakiyingi, Lydia; Sempa, Joseph; Mbabazi, Olive; Mukkada, Sheena; Nyesiga, Barnabas; Perkins, Mark D.; Manabe, Yukari C.

2013-01-01

Background Light emitting diode (LED) fluorescence microscopy (FM) is an affordable, technology targeted for use in resource-limited settings and recommended for widespread roll-out by the World Health Organization (WHO). We sought to compare the operational performance of three LED FM methods compared to light microscopy in a cohort of HIV-positive tuberculosis (TB) suspects at an urban clinic in a high TB burden country. Methods Two spot specimens collected from TB suspects were included in the study. Smears were stained using auramine O method and read after blinding by three LED-based FM methods by trained laboratory technicians in the Infectious Diseases Institutelaboratory. Leftover portions of the refrigerated sputum specimens were transported to the FIND Tuberculosis Research Laboratory for Ziehl Neelsen (ZN) smear preparation and reading by experienced technologist as well as liquid and solid culture. Results 174 of 627 (27.8%) specimens collected yielded one or more positive mycobacterial cultures. 94.3% (164/174) were M. tuberculosis complex. LED FM was between 7.3–11.0% more sensitive compared to ZN microscopy. Of the 592 specimens examined by all microscopy methods, there was no significant difference in sensitivity between the three LED FM methods. The specificity of the LED FM methods was between 6.1% and 7.7% lower than ZN microscopy (P<0.001), although exclusion of the single poor reader resulted in over 98% specificity for all FM methods. Conclusions Laboratory technicians in routine settings can be trained to use FM which is more sensitive than ZN microscopy. Despite rigorous proficiency testing, there were operator-dependent accuracy issues which highlight the critical need for intensive quality assurance procedures during LED FM implementation. The low sensitivity of FM for HIV-positive individuals particularly those with low CD4 T cell counts, will limit the number of additional patients found by LED FM in countries with high rates of HIV co-infection. PMID:24039780

Comparison of the Antibacterial Effect and Smear Layer Removal Using Photon-Initiated Photoacoustic Streaming Aided Irrigation Versus a Conventional Irrigation in Single-Rooted Canals: An In Vitro Study

PubMed Central

Zhu, Xiaofei; Yin, Xingzhe; Chang, Jeffrey W.W.; Wang, Yu; Cheung, Gary S.P.

2013-01-01

Abstract Objective: The Er:YAG laser with photon-induced photoacoustic streaming (PIPS) technique was reported to be effective in root canal disinfection. This study attempted to further investigate the antibacterial efficacy and smear layer removal ability of PIPS in comparison with conventional syringe irrigation in vitro. Methods: For antibacterial analysis, 48 single-rooted human teeth were prepared and inoculated with Enterococcus faecalis, and then divided into six groups of eight roots each. The colony-forming units (CFUs) per milliliter were determined after infection as the baseline. Then, the teeth were subjected to either PIPS plus 3% sodium hypochlorite (PIPS+NaOCl) or conventional syringe irrigation with 0.9% saline, 3% NaOCl, 17% ethylenediaminetetraacetic acid (EDTA), 0.2% chlorhexidine gluconate (CHX), or 3% NaOCl alternating with 17% EDTA. The reduction of CFUs in the individual group was determined. Additionally, scanning electron microscopy (SEM) examination of the canal walls for E. faecalis colonization was performed. For comparing the smear removal efficacy, another 48 single-rooted teeth, assigned to different groups as mentioned, were irrigated after mechanical instrumentation. The presence of a smear layer at different levels of the root canal was scored by SEM examination. Results: No significant differences were found in CFU reduction. No bacteria could be observed by SEM in the NaOCl, NaOCl+EDTA, and PIPS+NaOCl groups. The scores of smear layer of the NaOCl+EDTA and PIPS+NaOCl groups were significantly lower than those of the other groups in the coronal and middle third of the root canal. None of the methods can effectively remove smear layer in the apical third. Conclusions: PIPS system supplied with NaOCl and conventional syringe irrigation with NaOCl+EDTA are comparable in their ability to remove E. faecalis and smear layer in single-rooted canals. PMID:23863104

Utility of a Fluorescence Microscopy Imaging System for Analyzing the DNA Ploidy of Pathological Megakaryocytes Including 5q- Syndrome.

PubMed

Nakahara, Takako; Suemori, Shinichiro; Tsujioka, Takayuki; Kataoka, Mikio; Kataoka, Hiromi; Shibakura, Misako; Tohyama, Kaoru

2018-06-01

To investigate megakaryocyte (MK) DNA ploidy in various hematological diseases, fluorescence microscopy imaging system (FMI) can be used to analyze DNA ploidy with cell morphology at the single-cell level by using specialized image-processing software. Here we compared DNA ploidy obtained by FMI measured with that obtained flow cytometry (FCM). With FMI, we could evaluate the DNA ploidy in long-term preserved bone marrow smear samples after staining. We next analyzed the MK DNA ploidy in 42 bone marrow smear samples including 26 myeloid neoplasm cases, and we compared the DNA ploidy and platelet counts in the patients' peripheral blood; the production of platelets was significantly high compared to DNA ploidy in the myeloproliferative neoplasms group. The FMI method revealed that the patients with 5q- syndrome exhibited relatively low DNA ploidy despite high platelet counts, and this result suggested that increased DNA ploidy is not indispensable to abundant platelet production. The FMI method for DNA ploidy will be a useful tool to clarify the relationship between DNA ploidy and platelet production by MKs.

Efficiency of Different Endodontic Irrigation and Activation Systems in Removal of the Smear Layer: A Scanning Electron Microscopy Study.

PubMed

Karade, Priyatam; Chopade, Rutuja; Patil, Suvarna; Hoshing, Upendra; Rao, Madhukar; Rane, Neha; Chopade, Aditi; Kulkarni, Anish

2017-01-01

This in vitro study was designed to evaluate and compare different endodontic irrigation and activation systems for removal of the intracanal smear layer. Forty recently extracted, non-carious human intact single rooted premolars were selected and divided into five groups ( n =10) according to the root canal irrigation systems; syringe and needle irrigation (CTR), sonic irrigation, passive ultrasonic irrigation (PUI) and EndoVac irrigation system. All groups were prepared to #40 apical size with K-files. Each sample was subjected to final irrigation by using four different irrigation/activation systems. After splitting the samples, one half of each root was selected for examination under scanning electron microscope (SEM). The irrigation systems were compared using the Fisher's exact test with the level of significance set at 0.05. The four groups did not differ from each other in the coronal and mid-root parts of the canal. In the apical part of the canal none of the methods could completely remove all the smear layer but EndoVac system showed significantly better removal of smear layer and debris than the other methods. Within the limitations of the present study, the EndoVac system cleaned the apical part of the canal more efficiently than sonic, ultrasonic and syringe and needle irrigation.

Computer vision for microscopy diagnosis of malaria.

PubMed

Tek, F Boray; Dempster, Andrew G; Kale, Izzet

2009-07-13

This paper reviews computer vision and image analysis studies aiming at automated diagnosis or screening of malaria infection in microscope images of thin blood film smears. Existing works interpret the diagnosis problem differently or propose partial solutions to the problem. A critique of these works is furnished. In addition, a general pattern recognition framework to perform diagnosis, which includes image acquisition, pre-processing, segmentation, and pattern classification components, is described. The open problems are addressed and a perspective of the future work for realization of automated microscopy diagnosis of malaria is provided.

Theory-Informed Interventions to Improve the Quality of Tuberculosis Evaluation at Ugandan Health Centers: A Quasi-Experimental Study.

PubMed

Chaisson, Lelia H; Katamba, Achilles; Haguma, Priscilla; Ochom, Emmanuel; Ayakaka, Irene; Mugabe, Frank; Miller, Cecily; Vittinghoff, Eric; Davis, J Lucian; Handley, Margaret A; Cattamanchi, Adithya

2015-01-01

Tuberculosis (TB) remains under-diagnosed in many countries, in part due to poor evaluation practices at health facilities. Theory-informed strategies are needed to improve implementation of TB evaluation guidelines. We aimed to evaluate the impact of performance feedback and same-day smear microscopy on the quality of TB evaluation at 6 health centers in rural Uganda. We tested components of a multi-faceted intervention to improve adherence to the International Standards for Tuberculosis Care (ISTC): performance feedback and same-day smear microscopy. The strategies were selected based on a qualitative assessment guided by the Theory of Planned Behavior and the PRECEDE model. We collected patient data 6 months before and after the introduction of each intervention component, and compared ISTC adherence in the pre- and post-intervention periods for adults with cough ≥ 2 weeks' duration. The performance feedback evaluation included 1,446 adults; 838 (58%) were evaluated during the pre-intervention period and 608 (42%) during the post-intervention period. Performance feedback resulted in a 15% (95%CI +10% to +20%, p<0.001) increase in the proportion of patients receiving ISTC-adherent care. The same-day microscopy evaluation included 1,950 adults; 907 (47%) were evaluated during the pre-intervention period and 1,043 (53%) during the post-intervention period. Same-day microscopy was associated with a 14% (95%CI +10% to +18%, p<0.001) increase in the proportion of patients receiving ISTC-adherent care. Performance feedback and same-day microscopy should be considered along with ISTC training as part of a multi-faceted intervention to improve the quality of TB evaluation in other high TB burden countries.

Theory-Informed Interventions to Improve the Quality of Tuberculosis Evaluation at Ugandan Health Centers: A Quasi-Experimental Study

PubMed Central

Haguma, Priscilla; Ochom, Emmanuel; Ayakaka, Irene; Mugabe, Frank; Miller, Cecily; Vittinghoff, Eric; Davis, J. Lucian; Handley, Margaret A.; Cattamanchi, Adithya

2015-01-01

Background Tuberculosis (TB) remains under-diagnosed in many countries, in part due to poor evaluation practices at health facilities. Theory-informed strategies are needed to improve implementation of TB evaluation guidelines. We aimed to evaluate the impact of performance feedback and same-day smear microscopy on the quality of TB evaluation at 6 health centers in rural Uganda. Methods We tested components of a multi-faceted intervention to improve adherence to the International Standards for Tuberculosis Care (ISTC): performance feedback and same-day smear microscopy. The strategies were selected based on a qualitative assessment guided by the Theory of Planned Behavior and the PRECEDE model. We collected patient data 6 months before and after the introduction of each intervention component, and compared ISTC adherence in the pre- and post-intervention periods for adults with cough ≥ 2 weeks’ duration. Results The performance feedback evaluation included 1,446 adults; 838 (58%) were evaluated during the pre-intervention period and 608 (42%) during the post-intervention period. Performance feedback resulted in a 15% (95%CI +10% to +20%, p<0.001) increase in the proportion of patients receiving ISTC-adherent care. The same-day microscopy evaluation included 1,950 adults; 907 (47%) were evaluated during the pre-intervention period and 1,043 (53%) during the post-intervention period. Same-day microscopy was associated with a 14% (95%CI +10% to +18%, p<0.001) increase in the proportion of patients receiving ISTC-adherent care. Conclusions Performance feedback and same-day microscopy should be considered along with ISTC training as part of a multi-faceted intervention to improve the quality of TB evaluation in other high TB burden countries. PMID:26172948

Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chaikuad, Apirat, E-mail: apirat.chaikuad@sgc.ox.ac.uk; Knapp, Stefan; Johann Wolfgang Goethe-University, Building N240 Room 3.03, Max-von-Laue-Strasse 9, 60438 Frankfurt am Main

An alternative strategy for PEG sampling is suggested through the use of four newly defined PEG smears to enhance chemical space in reduced screens with a benefit towards protein crystallization. The quest for an optimal limited set of effective crystallization conditions remains a challenge in macromolecular crystallography, an issue that is complicated by the large number of chemicals which have been deemed to be suitable for promoting crystal growth. The lack of rational approaches towards the selection of successful chemical space and representative combinations has led to significant overlapping conditions, which are currently present in a multitude of commercially availablemore » crystallization screens. Here, an alternative approach to the sampling of widely used PEG precipitants is suggested through the use of PEG smears, which are mixtures of different PEGs with a requirement of either neutral or cooperatively positive effects of each component on crystal growth. Four newly defined smears were classified by molecular-weight groups and enabled the preservation of specific properties related to different polymer sizes. These smears not only allowed a wide coverage of properties of these polymers, but also reduced PEG variables, enabling greater sampling of other parameters such as buffers and additives. The efficiency of the smear-based screens was evaluated on more than 220 diverse recombinant human proteins, which overall revealed a good initial crystallization success rate of nearly 50%. In addition, in several cases successful crystallizations were only obtained using PEG smears, while various commercial screens failed to yield crystals. The defined smears therefore offer an alternative approach towards PEG sampling, which will benefit the design of crystallization screens sampling a wide chemical space of this key precipitant.« less

Evaluation of the smear layer removal and erosive capacity of EDTA, boric acid, citric acid and desy clean solutions: an in vitro study.

PubMed

Turk, Tugba; Kaval, Mehmet Emin; Şen, Bilge Hakan

2015-09-03

The purpose of this study was to investigate the smear layer removal and erosive capacity of various irrigation solutions with sequential use of NaOCl on instrumented root canal walls. The root canals of single-rooted teeth were instrumented with ProTaper rotary instrument. Then, the teeth were randomly divided into five experimental groups. The root canals were irrigated with one of the following solutions (5 mL/1 min): 5% EDTA, 5% boric acid (BA), a mixture of BA and CA, 2.5% citric acid (CA) and 5% Desy Clean. After irrigating with 2.5% NaOCl and distilled water, the roots were split into two halves and each half was prepared for SEM examination. Representative photographs were taken from each third at x500 and x1000 magnifications. Double blind scoring was performed by two calibrated observers for smear layer and erosion. The scores were statistically analyzed using Kruskal-Wallis, Dunn's post hoc and Spearman's correlation tests (p = 0.05). There were statistically significant differences among the solutions by means of smear layer and erosion (p < 0.05). While 2.5% CA solution was the most effective solution in removal of smear layer, it was also the most erosive solution (p < 0.05). 5% Desy Clean removed smear layer effectively and caused less erosion. There was a negative, but statistically significant correlation between presence of smear layer and erosion (r = -0.684; p < 0.0001). Desy Clean can be a promising agent as an irrigation solution with optimal smear layer removal capacity and less erosive effects.

Effect of Active Case Finding on Prevalence and Transmission of Pulmonary Tuberculosis in Dhaka Central Jail, Bangladesh

PubMed Central

Banu, Sayera; Rahman, Md. Toufiq; Uddin, Mohammad Khaja Mafij; Khatun, Razia; Khan, Md. Siddiqur Rahman; Rahman, Md. Mojibur; Uddin, Syed Iftekhar; Ahmed, Tahmeed; Heffelfinger, James D.

2015-01-01

Background Understanding tuberculosis (TB) transmission dynamics is essential for establishing effective TB control strategies in settings where the burden and risk of transmission are high. The objectives of this study were to evaluate the effect of active screening on controlling TB transmission and also to characterize Mycobacterium tuberculosis strains for investigating transmission dynamics in a correctional setting. Methods The study was carried out in Dhaka Central Jail (DCJ), from October 2005 to February 2010. An active case finding strategy for pulmonary TB was established both at the entry point to the prison and inside the prison. Three sputum specimens were collected from all pulmonary TB suspects and subjected to smear microscopy, culture, and drug susceptibility testing as well as genotyping which included deletion analysis, spoligotyping and analysis of mycobacterial interspersed repetitive units (MIRU). Results A total of 60,585 inmates were screened during the study period. We found 466 inmates with pulmonary TB of whom 357 (77%) had positive smear microscopy results and 109 (23%) had negative smear microscopy results but had positive results on culture. The number of pulmonary TB cases declined significantly, from 49 cases during the first quarter to 8 cases in the final quarter of the study period (p=0.001). Deletion analysis identified all isolates as M. tuberculosis and further identified 229 (70%) strains as ‘modern’ and 100 (30%) strains as ‘ancestral’. Analysis of MIRU showed that 347 strains (85%) exhibited unique patterns, whereas 61 strains (15%) clustered into 22 groups. The largest cluster comprised eight strains of the Beijing M. tuberculosis type. The rate of recent transmission was estimated to be 9.6%. Conclusions Implementation of active screening for TB was associated with a decline in TB cases in DCJ. Implementation of active screening in prison settings might substantially reduce the national burden of TB in Bangladesh. PMID:25933377

Effect of active case finding on prevalence and transmission of pulmonary tuberculosis in Dhaka Central Jail, Bangladesh.

PubMed

Banu, Sayera; Rahman, Md Toufiq; Uddin, Mohammad Khaja Mafij; Khatun, Razia; Khan, Md Siddiqur Rahman; Rahman, Md Mojibur; Uddin, Syed Iftekhar; Ahmed, Tahmeed; Heffelfinger, James D

2015-01-01

Understanding tuberculosis (TB) transmission dynamics is essential for establishing effective TB control strategies in settings where the burden and risk of transmission are high. The objectives of this study were to evaluate the effect of active screening on controlling TB transmission and also to characterize Mycobacterium tuberculosis strains for investigating transmission dynamics in a correctional setting. The study was carried out in Dhaka Central Jail (DCJ), from October 2005 to February 2010. An active case finding strategy for pulmonary TB was established both at the entry point to the prison and inside the prison. Three sputum specimens were collected from all pulmonary TB suspects and subjected to smear microscopy, culture, and drug susceptibility testing as well as genotyping which included deletion analysis, spoligotyping and analysis of mycobacterial interspersed repetitive units (MIRU). A total of 60,585 inmates were screened during the study period. We found 466 inmates with pulmonary TB of whom 357 (77%) had positive smear microscopy results and 109 (23%) had negative smear microscopy results but had positive results on culture. The number of pulmonary TB cases declined significantly, from 49 cases during the first quarter to 8 cases in the final quarter of the study period (p=0.001). Deletion analysis identified all isolates as M. tuberculosis and further identified 229 (70%) strains as 'modern' and 100 (30%) strains as 'ancestral'. Analysis of MIRU showed that 347 strains (85%) exhibited unique patterns, whereas 61 strains (15%) clustered into 22 groups. The largest cluster comprised eight strains of the Beijing M. tuberculosis type. The rate of recent transmission was estimated to be 9.6%. Implementation of active screening for TB was associated with a decline in TB cases in DCJ. Implementation of active screening in prison settings might substantially reduce the national burden of TB in Bangladesh.

Total variation based image deconvolution for extended depth-of-field microscopy images

NASA Astrophysics Data System (ADS)

Hausser, F.; Beckers, I.; Gierlak, M.; Kahraman, O.

2015-03-01

One approach for a detailed understanding of dynamical cellular processes during drug delivery is the use of functionalized biocompatible nanoparticles and fluorescent markers. An appropriate imaging system has to detect these moving particles so as whole cell volumes in real time with high lateral resolution in a range of a few 100 nm. In a previous study Extended depth-of-field microscopy (EDF-microscopy) has been applied to fluorescent beads and tradiscantia stamen hair cells and the concept of real-time imaging has been proved in different microscopic modes. In principle a phase retardation system like a programmable space light modulator or a static waveplate is incorporated in the light path and modulates the wavefront of light. Hence the focal ellipsoid is smeared out and images seem to be blurred in a first step. An image restoration by deconvolution using the known point-spread-function (PSF) of the optical system is necessary to achieve sharp microscopic images of an extended depth-of-field. This work is focused on the investigation and optimization of deconvolution algorithms to solve this restoration problem satisfactorily. This inverse problem is challenging due to presence of Poisson distributed noise and Gaussian noise, and since the PSF used for deconvolution exactly fits in just one plane within the object. We use non-linear Total Variation based image restoration techniques, where different types of noise can be treated properly. Various algorithms are evaluated for artificially generated 3D images as well as for fluorescence measurements of BPAE cells.

[Advances in automatic detection technology for images of thin blood film of malaria parasite].

PubMed

Juan-Sheng, Zhang; Di-Qiang, Zhang; Wei, Wang; Xiao-Guang, Wei; Zeng-Guo, Wang

2017-05-05

This paper reviews the computer vision and image analysis studies aiming at automated diagnosis or screening of malaria in microscope images of thin blood film smears. On the basis of introducing the background and significance of automatic detection technology, the existing detection technologies are summarized and divided into several steps, including image acquisition, pre-processing, morphological analysis, segmentation, count, and pattern classification components. Then, the principles and implementation methods of each step are given in detail. In addition, the promotion and application in automatic detection technology of thick blood film smears are put forwarded as questions worthy of study, and a perspective of the future work for realization of automated microscopy diagnosis of malaria is provided.

Morphological analysis of red blood cells by polychromatic interference microscopy of thin films

NASA Astrophysics Data System (ADS)

Dyachenko, A. A.; Malinova, L. I.; Ryabukho, V. P.

2016-11-01

Red blood cells (RBC) distribution width (RDW) is a promising hematological parameter with broadapplications in clinical practice; in various studies RDWhas been shown to be associated with increased risk of heart failure (HF) in general population. It predicts mortality and other major adverse events in HF patients. In this report new method of RDWmeasurement is presented. It's based on interference color analysis of red blood cells in blood smear and further measurement of its optical thickness. Descriptive statistics of the of the RBC optical thickness distribution in a blood smear were used for RDW estimation in every studied sample. Proposed method is considered to be avoiding type II errors and minimizing the variability of measured RDW.

Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens

PubMed Central

Chaikuad, Apirat; Knapp, Stefan; von Delft, Frank

2015-01-01

The quest for an optimal limited set of effective crystallization conditions remains a challenge in macromolecular crystallography, an issue that is complicated by the large number of chemicals which have been deemed to be suitable for promoting crystal growth. The lack of rational approaches towards the selection of successful chemical space and representative combinations has led to significant overlapping conditions, which are currently present in a multitude of commercially available crystallization screens. Here, an alternative approach to the sampling of widely used PEG precipitants is suggested through the use of PEG smears, which are mixtures of different PEGs with a requirement of either neutral or cooperatively positive effects of each component on crystal growth. Four newly defined smears were classified by molecular-weight groups and enabled the preservation of specific properties related to different polymer sizes. These smears not only allowed a wide coverage of properties of these polymers, but also reduced PEG variables, enabling greater sampling of other parameters such as buffers and additives. The efficiency of the smear-based screens was evaluated on more than 220 diverse recombinant human proteins, which overall revealed a good initial crystallization success rate of nearly 50%. In addition, in several cases successful crystallizations were only obtained using PEG smears, while various commercial screens failed to yield crystals. The defined smears therefore offer an alternative approach towards PEG sampling, which will benefit the design of crystallization screens sampling a wide chemical space of this key precipitant. PMID:26249344

Defined PEG smears as an alternative approach to enhance the search for crystallization conditions and crystal-quality improvement in reduced screens.

PubMed

Chaikuad, Apirat; Knapp, Stefan; von Delft, Frank

2015-08-01

The quest for an optimal limited set of effective crystallization conditions remains a challenge in macromolecular crystallography, an issue that is complicated by the large number of chemicals which have been deemed to be suitable for promoting crystal growth. The lack of rational approaches towards the selection of successful chemical space and representative combinations has led to significant overlapping conditions, which are currently present in a multitude of commercially available crystallization screens. Here, an alternative approach to the sampling of widely used PEG precipitants is suggested through the use of PEG smears, which are mixtures of different PEGs with a requirement of either neutral or cooperatively positive effects of each component on crystal growth. Four newly defined smears were classified by molecular-weight groups and enabled the preservation of specific properties related to different polymer sizes. These smears not only allowed a wide coverage of properties of these polymers, but also reduced PEG variables, enabling greater sampling of other parameters such as buffers and additives. The efficiency of the smear-based screens was evaluated on more than 220 diverse recombinant human proteins, which overall revealed a good initial crystallization success rate of nearly 50%. In addition, in several cases successful crystallizations were only obtained using PEG smears, while various commercial screens failed to yield crystals. The defined smears therefore offer an alternative approach towards PEG sampling, which will benefit the design of crystallization screens sampling a wide chemical space of this key precipitant.

Application of image recognition-based automatic hyphae detection in fungal keratitis.

PubMed

Wu, Xuelian; Tao, Yuan; Qiu, Qingchen; Wu, Xinyi

2018-03-01

The purpose of this study is to evaluate the accuracy of two methods in diagnosis of fungal keratitis, whereby one method is automatic hyphae detection based on images recognition and the other method is corneal smear. We evaluate the sensitivity and specificity of the method in diagnosis of fungal keratitis, which is automatic hyphae detection based on image recognition. We analyze the consistency of clinical symptoms and the density of hyphae, and perform quantification using the method of automatic hyphae detection based on image recognition. In our study, 56 cases with fungal keratitis (just single eye) and 23 cases with bacterial keratitis were included. All cases underwent the routine inspection of slit lamp biomicroscopy, corneal smear examination, microorganism culture and the assessment of in vivo confocal microscopy images before starting medical treatment. Then, we recognize the hyphae images of in vivo confocal microscopy by using automatic hyphae detection based on image recognition to evaluate its sensitivity and specificity and compare with the method of corneal smear. The next step is to use the index of density to assess the severity of infection, and then find the correlation with the patients' clinical symptoms and evaluate consistency between them. The accuracy of this technology was superior to corneal smear examination (p < 0.05). The sensitivity of the technology of automatic hyphae detection of image recognition was 89.29%, and the specificity was 95.65%. The area under the ROC curve was 0.946. The correlation coefficient between the grading of the severity in the fungal keratitis by the automatic hyphae detection based on image recognition and the clinical grading is 0.87. The technology of automatic hyphae detection based on image recognition was with high sensitivity and specificity, able to identify fungal keratitis, which is better than the method of corneal smear examination. This technology has the advantages when compared with the conventional artificial identification of confocal microscope corneal images, of being accurate, stable and does not rely on human expertise. It was the most useful to the medical experts who are not familiar with fungal keratitis. The technology of automatic hyphae detection based on image recognition can quantify the hyphae density and grade this property. Being noninvasive, it can provide an evaluation criterion to fungal keratitis in a timely, accurate, objective and quantitative manner.

Evaluation of Blood Cell Attachment on Er:Yag Laser Applied Root Surface Using Scanning Electron Microscopy

PubMed Central

CEKICI, Ali; MADEN, Ilay; YILDIZ, Sercan; SAN, Tangul; ISIK, Gulden

2013-01-01

Background: Periodontal regeneration is dependent on the uninterrupted adhesion, maturation and absorption of fibrin clots to a periodontally compromised root surface. The modification of the root surface with different agents has been used for better fibrin clot formation and blood cell attachment. It is known that Er:YAG laser application on dentin removes the smear layer succesfully. Aim: The aim of this study is to observe blood cell attachment and fibrin network formation following ER:YAG laser irradiation on periodontally compromised root surfaces in comparison to chemical root conditioning techniques in vitro. Materials and methods: 40 dentin blocks prepared from freshly extracted periodontally compromised hopeless teeth. Specimens were divided in 5 groups; those applied with PBS, EDTA, Citric acid and Er:YAG. They were further divided into two groups: those which had received these applications, and the control group. The specimens were evaluated with scanning electron microscope and micrographs were taken. Smear layer and blood cell attachment scoring was performed. Results: In the Er:YAG laser applied group, smear layer were totally removed. In the blood applied specimens, better fibrin clot formation and blood cell attachment were observed in the Er:YAG group. In the group that had been applied with citric acid, the smear layer was also removed. The smear layer could not be fully removed in the EDTA group. Conclusion: Er:YAG laser application on the root dentin seems to form a suitable surface for fibrin clot formation and blood cell attachment. Further clinical studies to support these results are necessitated. PMID:23533017

Image classification of unlabeled malaria parasites in red blood cells.

PubMed

Zheng Zhang; Ong, L L Sharon; Kong Fang; Matthew, Athul; Dauwels, Justin; Ming Dao; Asada, Harry

2016-08-01

This paper presents a method to detect unlabeled malaria parasites in red blood cells. The current "gold standard" for malaria diagnosis is microscopic examination of thick blood smear, a time consuming process requiring extensive training. Our goal is to develop an automate process to identify malaria infected red blood cells. Major issues in automated analysis of microscopy images of unstained blood smears include overlapping cells and oddly shaped cells. Our approach creates robust templates to detect infected and uninfected red cells. Histogram of Oriented Gradients (HOGs) features are extracted from templates and used to train a classifier offline. Next, the ViolaJones object detection framework is applied to detect infected and uninfected red cells and the image background. Results show our approach out-performs classification approaches with PCA features by 50% and cell detection algorithms applying Hough transforms by 24%. Majority of related work are designed to automatically detect stained parasites in blood smears where the cells are fixed. Although it is more challenging to design algorithms for unstained parasites, our methods will allow analysis of parasite progression in live cells under different drug treatments.

Effectiveness of Rotary Endodontic Instruments on Smear Layer Removal in Root Canals of Primary Teeth: A Scanning Electron Microscopy Study.

PubMed

Subramaniam, Priya; Girish Babu, K L; Tabrez, T A

2016-01-01

The present SEM study was undertaken to evaluate the effect of root canal instrumentation using both manual and rotary files in the root canals of primary anterior teeth. Thirty freshly extracted primary maxillary incisors were divided into 3 groups of 10 teeth each. In Group I, root canals were instrumented with rotary NiTi files; in Group II, the root canals were instrumented using manual NiTi K files and; in Group III, manual instrumentation was done with stainless steel K files. Longitudinal sections were prepared and processed for observation under SEM at the coronal, middle and apical thirds. Scoring of smear layer was done according to Hulsmann and the data obtained was subjected to statistical analysis. Rotary files cleaned the coronal and middle thirds of root canals more effectively. Statistically there was no significant difference between the groups. Lowest score of 2.6 in the apical third of root canals was seen with hand NiTi files. Rotary instrumentation was as effective as manual instrumentation in removal of smear layer in the root canals of primary anterior teeth.

Evaluation of genotype MTBDRplus VER 2.0 line probe assay for the detection of MDR-TB in smear positive and negative sputum samples.

PubMed

Meaza, Abyot; Kebede, Abebaw; Yaregal, Zelalem; Dagne, Zekarias; Moga, Shewki; Yenew, Bazezew; Diriba, Getu; Molalign, Helina; Tadesse, Mengistu; Adisse, Desalegn; Getahun, Muluwork; Desta, Kassu

2017-04-17

Multi drug resistant tuberculosis (MDR-TB) poses formidable challenges to TB control due to its complex diagnostic and treatment challenges and often associated with a high rate of mortality. Accurate and rapid detection of MDR-TB is critical for timely initiation of treatment. Line Probe Assay (LPA) is a qualitative in vitro diagnostic test based on DNA-STRIP technology for the identification of the M. tuberculosis complex and its resistance to rifampicin (RMP) and/or isoniazid (INH). Hain Lifescience, GmbH, Germany has improved the sensitivity of Genotype MTBDRplus VER 2.0 LPA for the detection of MDR-TB; with the possibility of applying the tool in smear negative sputum samples. A cross sectional study was conducted on 274 presumptive MDR-TB patients referred to the National TB Reference Laboratory (NTRL), Ethiopian Public Health Institute (EPHI) who submitted sputum samples for laboratory diagnosis of drug resistant-TB testing. Seventy-two smear and culture positive samples processed in smear positive direct LPA category and 197 smear negative sputum samples were processed for direct LPA. Among the smear negative samples 145 (73.6%) were culture negative and 26 (13.2%) were culture positive. All specimens were processed using NALC-NaOH method and ZN smear microscopy done from sediments. Genotype MTBDRplus VER 2.0 done from processed sputum sediments and the result was compared against the reference, BACTEC MGIT 960 culture and DST. Sensitivity, specificity, PPV and NPV of Genotype MTBDRplus VER 2.0 assay was determined and P-value <0.05 was considered as statistically significant. The sensitivity, specificity, PPV and NPV of Genotype MTBDRplus VER 2.0 LPA were 96.4, 100, 100 and 96.9%, respectively for the detection of MDR-TB from direct smear positive sputum samples. The sensitivity, specificity, PPV and NPV of Genotype MTBDR plus VER 2.0 LPA were 77.8, 97.2, 82.4 and 97.2%, respectively, for the detection of M. tuberculosis from direct smear negative sputum samples. Fourteen (53.8%) samples had valid results with LPA among the 26 smear negative culture positive samples. The remaining 8 (30.8%) and 4 (15.4%) were invalid and negative with LPA, respectively. The sensitivity and specificity of Genotype MTBDRplus VER 2.0 LPA were 100% for the detection of MDR-TB among 14 direct smear negative and culture positive sputum samples. The most common mutations associated with RMP and INH resistance were S531L and S315TL, respectively. A single rare mutation (C15T/A16G) was detected for INH resistance. The diagnostic performance of Genotype MTBDRplus VER 2.0 LPA in direct smear positive sputum sample was highly sensitive and specific for early detection of MDR-TB. However, the diagnostic performance of this molecular assay in direct smear negative sputum sample was low and showed a high level of invalid results for detection of M. tuberculosis and its resistance to RMP and/or INH so it is unlikely to implement Genotype MTBDRplus VER 2.0 for the detection of MDR-TB in direct smear negative sample in our routine settings. The sensitivity of the assay should be improved for detection of MDR-TB in direct smear negative sputum specimens.

Adhesion of Mycobacterium smegmatis to Charged Surfaces and Diagnostics Implications

NASA Astrophysics Data System (ADS)

Gorse, Diane; Dhinojwala, Ali; Moore, Francisco

Pulmonary tuberculosis (PTB) causes more than 1 million deaths annually. Smear microscopy is a primary rapid detection tool in areas where 95 % of PTB cases occur. This technique, in which the sputum of a symptomatic patient is stained and examined using a light microscope for Mycobacterium tuberculosis (MTB) shows sensitivity between 20 and 60 %. Insufficient bacterial isolation during sample preparation may be a reason for low sensitivity. We are optimizing a system to capture bacteria on the basis of electrostatic interactions to more thoroughly isolate bacteria from suspension and facilitate more accurate detection. Silica supports coated with positively-charged polyelectrolyte, poly(diallyldimethylammonium chloride), captured approximately 4.1 times more Mycobacterium smegmatis, a model organism for MTB, than was captured on negatively-charged silica substrates. Future experimentation will employ branched polymer systems and seek to justify the use of colloidal stability theories to describe initial capture. Supported by University of Akron, Department of Polymer Science, Department of Biology; LORD Corporation.

[Microbiological diagnosis of gas gangrene caused by Clostridium septicum (a clinical case)].

PubMed

Men'shikova, E D; Titova, G P; Kartavenko, V I; Sokolov, V A; Shabanov, A K; Men'shikov, D D

2010-08-01

Microscopy of gram-stained impression smears is used for the rapid diagnosis of microorganisms in the wound. The shin tissues of patient P. with suspected gas gangrene of lower extremity soft tissues were microscopically found to have gram-positive spore-forming bacteria that were morphologically similar to C. bifermentans that were identified as C. septicum on cultural diagnosis. The pathogenic C. septicum strain spores were likely to be formed in the macroorganism upon exposure of the pathogen to a patient's defense factors and to a package of therapeutic measures. Microbiological data should be used only in combination with clinical and instrumental findings and the results of other laboratory studies when the optimal technology is chosen to treat gas infection. By keeping in mind that there may be clostridial gangrene in the patients and the experience of clinicians and bacteriologists may be insufficient in diagnosing this pathology, it is necessary to strengthen the training of physicians in the diagnosis of this pathology.

A comparative scanning electron microscopic study of the effect of three different rotary instruments on smear layer formation.

PubMed

Shahi, Shahriar; Yavari, Hamid R; Rahimi, Saeed; Reyhani, Mohammad F; Kamarroosta, Zahra; Abdolrahimi, Majid

2009-03-01

The aim of this study was to evaluate the effect of RaCe, FlexMaster and ProFile rotary instruments on smear layer formation by scanning electron microscopy. Eighty-four caries-free freshly extracted human single-rooted teeth were selected and divided into three groups, each containing 28 teeth. The teeth were instrumented with rotary instruments sequentially: Group A: ProFile Rotary Instruments; Group B: FlexMaster Rotary Instruments; and Group C: RaCe Rotary Instruments. Instrumentation was performed by the crown-down method and according to the manufacturer's instructions. The specimens were then examined with SEM according to Hülsmann's classification. One-way ANOVA and a post hoc Tukey test were used for statistical analysis. The results showed that there were no statistically significant differences among the three groups in the coronal third (P = 0.39), but at the apical and middle thirds there were statistically significant differences between the RaCe group and the other groups (P < 0.05). Smear layer in the RaCe group was less than that in the ProFile and FlexMaster groups, but the difference between the ProFile group and FlexMaster group was not statistically significant (P > 0.05). It was concluded that RaCe Rotary Instruments produce less smear layer than FlexMaster and ProFile Rotary Instruments.

Sputum, sex and scanty smears: new case definition may reduce sex disparities in smear-positive tuberculosis.

PubMed

Ramsay, A; Bonnet, M; Gagnidze, L; Githui, W; Varaine, F; Guérin, P J

2009-05-01

Urban clinic, Nairobi. To evaluate the impact of specimen quality and different smear-positive tuberculosis (TB) case (SPC) definitions on SPC detection by sex. Prospective study among TB suspects. A total of 695 patients were recruited: 644 produced > or =1 specimen for microscopy. The male/female sex ratio was 0.8. There were no significant differences in numbers of men and women submitting three specimens (274/314 vs. 339/380, P = 0.43). Significantly more men than women produced a set of three 'good' quality specimens (175/274 vs. 182/339, P = 0.01). Lowering thresholds for definitions to include scanty smears resulted in increases in SPC detection in both sexes; the increase was significantly higher for women. The revised World Health Organization (WHO) case definition was associated with the highest detection rates in women. When analysis was restricted only to patients submitting 'good' quality specimen sets, the difference in detection between sexes was on the threshold for significance (P = 0.05). Higher SPC notification rates in men are commonly reported by TB control programmes. The revised WHO SPC definition may reduce sex disparities in notification. This should be considered when evaluating other interventions aimed at reducing these. Further study is required on the effects of the human immuno-deficiency virus and instructed specimen collection on sex-specific impact of new SPC definition.

[Comparison of the efficacies of cross priming amplification and RealAmp with XpertMTB/RIF for the diagnosis of pulmonary tuberculosis at peripheral microscopic center].

PubMed

Wang, S H; Zheng, D W; Zhu, Y K; Ma, X G; Shi, J; Ou, X C; Li, H; Xing, J; Zhao, Y L

2018-02-12

Objective: To compare the efficacies of cross priming amplification (CPA) and RealAmp with XpertMTB/RIF for the diagnosis of pulmonary tuberculosis(TB) at peripheral microscopic centers. Methods: From December of 2014 to December of 2015, 3 193 patients suspected with TB were enrolled consecutively at 3 county level TB clinical clinics in Zhongmu, Xinmi and Dengzhou of Henan province. Totally 3 193 collected sputum samples were detected by smear microscopy, L-J media culture, CPA, RealAmp and Xpert MTB/RIF. The culture positive samples were tested by MPB64 for strain identification. The sensitivity and specificity of CPA, RealAmp and Xpert MTB/RIF were calculated according to L-J solid culture results and clinical diagnosis results. Results: The sensitivity of CPA, RealAmp and Xpert MTB/RIF were 85.5%(413/483), 85.5%(413/483) and 87.9%(422/480), respectively, compared with L-J solid culture, the difference among the 3 methods being not significant(χ(2)=1.6, P >0.05). The specificity of CPA, RealAmp and Xpert MTB/RIF were 96.8%(2 624/2 170), 93.2%(2 527/2 170) and 95.3%(2 567/2 170) compared with culture; and there was a significantly statistic difference among the 3 methods(χ(2)=37.8, P <0.001). The sensitivity of smear microscopy, culture, CPA, RealAmp and Xpert MTB/RIF was 21.7%(300/1 383), 34.9%(483/1 383), 34.6%(478/1 383), 39.2%(542/1 383) and 38.1%(526/1 381) compared with clinical diagnosis. The sensitivity of CPA, RealAmp and Xpert MTB/RIF was higher than that of smear (χ(2) =31.9, P <0.01), but there was no significantly statistic difference between the 3 molecular methods(χ(2)=2.9, P >0.05). The specificity of smear microscopy, L-J solid culture, CPA, RealAmp and Xpert MTB/RIF was 100%(1 810/1 810), 100%(1 810/1 810), 98.8%(1 789/1 810), 98.8%(1 756/1 810) and 97.0%(1 788/1 810), and there was no significantly statistic difference among the 3 molecular methods(χ(2)=0.16, P >0.05). Conclusion: The capability of CPA and RealAmp for diagnosing pulmonary TB was similar to Xpert MTB/RIF.The former 2 methods were more suitable to apply to the diagnoses of pulmonary TB in peripheral laboratories.

Mobile-Based Analysis of Malaria-Infected Thin Blood Smears: Automated Species and Life Cycle Stage Determination.

PubMed

Rosado, Luís; da Costa, José M Correia; Elias, Dirk; Cardoso, Jaime S

2017-09-21

Microscopy examination has been the pillar of malaria diagnosis, being the recommended procedure when its quality can be maintained. However, the need for trained personnel and adequate equipment limits its availability and accessibility in malaria-endemic areas. Rapid, accurate, accessible diagnostic tools are increasingly required, as malaria control programs extend parasite-based diagnosis and the prevalence decreases. This paper presents an image processing and analysis methodology using supervised classification to assess the presence of malaria parasites and determine the species and life cycle stage in Giemsa-stained thin blood smears. The main differentiation factor is the usage of microscopic images exclusively acquired with low cost and accessible tools such as smartphones, a dataset of 566 images manually annotated by an experienced parasilogist being used. Eight different species-stage combinations were considered in this work, with an automatic detection performance ranging from 73.9% to 96.2% in terms of sensitivity and from 92.6% to 99.3% in terms of specificity. These promising results attest to the potential of using this approach as a valid alternative to conventional microscopy examination, with comparable detection performances and acceptable computational times.

Usefulness of an inexpensive, Paracheck test in detecting asymptomatic infectious reservoir of plasmodium falciparum during dry season in an inaccessible terrain in central India.

PubMed

Singh, N; Saxena, A; Sharma, V P

2002-10-01

The performance of a new indigenous rapid diagnostic test, Paracheck Pf was evaluated in detection of Plasmodium falciparum in asymptomatic children in remote forest villages of Mandla district, central India to determine the lower limits of sensitivity and specificity of rapid test. A finger prick blood sample was collected to prepare blood smear and for testing with the Paracheck test. The blood smears were read by an experienced technician blinded to the Paracheck results. The figures for specificity, sensitivity, accuracy and predictive values were calculated using microscopy as gold standard. The new diagnostic test had a sensitivity of 94% and a specificity of 89%. The positive and negative predictive values were 71% and 98%, respectively. The J -index was 0.83%. The rapid test was found to be very easy to perform and the result could be read reliably by field workers. The field evaluation with this new inexpensive test, ($0.65/test) indicates that it could be used as an epidemiological tool in the management of malaria particularly in areas where microscopy is not operationally feasible to attain the goal of the roll back malaria initiative.

Mobile-Based Analysis of Malaria-Infected Thin Blood Smears: Automated Species and Life Cycle Stage Determination

PubMed Central

da Costa, José M. Correia; Elias, Dirk

2017-01-01

Microscopy examination has been the pillar of malaria diagnosis, being the recommended procedure when its quality can be maintained. However, the need for trained personnel and adequate equipment limits its availability and accessibility in malaria-endemic areas. Rapid, accurate, accessible diagnostic tools are increasingly required, as malaria control programs extend parasite-based diagnosis and the prevalence decreases. This paper presents an image processing and analysis methodology using supervised classification to assess the presence of malaria parasites and determine the species and life cycle stage in Giemsa-stained thin blood smears. The main differentiation factor is the usage of microscopic images exclusively acquired with low cost and accessible tools such as smartphones, a dataset of 566 images manually annotated by an experienced parasilogist being used. Eight different species-stage combinations were considered in this work, with an automatic detection performance ranging from 73.9% to 96.2% in terms of sensitivity and from 92.6% to 99.3% in terms of specificity. These promising results attest to the potential of using this approach as a valid alternative to conventional microscopy examination, with comparable detection performances and acceptable computational times. PMID:28934170

THE ROLE OF TARGET ORGAN DIAGNOSTIC APPROACH IN SEASONAL ALLERGIC RHINITIS: NASAL SMEAR EOSINOPHILS.

PubMed

Nurkic, Jasmina; Ahmad, Mona Al; Arifhodzic, Nermina; Jusufovic, Edin

2016-04-01

Allergic rhinitis (AR) related to local weeds pollen sensitization (Chenopodiaceous family) is the most common cause of respiratory allergy in Kuwait. Local nasal accumulation of different cells typical of allergic inflammation is responsible for clinical symptoms of AR. Although nasal smear for Eosinophils (NSE) is one of the earliest included valuable test in diagnosis of AR, with time is underestimated. Explore possible correlation of natural pollen allergen stimulation with appearance and quantity of Eosinophils in nasal smear. A group of randomly selected patients with clinical history suggestive for seasonal AR (SAR), who came to Al Rashed Allergy Center in period from October 2014 to October 2015, obtain Nasal Smear for Eosinophils as a screening test before further diagnostic evaluation. Nasal samples were collected by passing a sterile swab, from each nasal cavity, along the medial surface of the inferior turbinate 2 to 3 times and the specimen smeared on a clear glass slide. Nasal smears were examined by light microscopy after staining with hematoxylin and eosin stain. Skin prick test is performed in all symptomatic patients with a battery of inhalant allergens that include local pollens. The control group was recruited, with their voluntary consent, from the medical stuff with a negative history of any allergic nasal symptoms. In this group we performed only nasal smear for Eosinophils. Air Biology Laboratory Kuwait provided us with daily pollen count. From total 158 study participants, 132 had SAR symptoms and are divided in four groups. Fifth, control, group is non symptomatic. For 38.6% of symptomatic patients NSE were positive, while 45% of these patients have negative SPT. From 62.1% NSE negative patients, 37.8% have negative SPT. Our results showed expected positive correlation of NSE positive patients with pollen season in Kuwait, in SPT positive group. However, presence of Eosinophils in nasal smear was moderate to high also in patients with negative SPT during the highest peak of season, in contrast to control group. NES showed moderate sensitivity, relatively high specificity and importance as screening test in SPT negative patients. Evaluation of AR demand wide and improved diagnostic approach due to significant number of SPT negative patients with positive NSE based on natural allergen stimulation. Our results emphasize locale allergic response of nasal mucosa and importance of target organ diagnostic approach.

Comparison of positive-pressure, passive ultrasonic, and laser-activated irrigations on smear-layer removal from the root canal surface.

PubMed

Sahar-Helft, Sharonit; Sarp, Ayşe Sena Kabaş; Stabholtz, Adam; Gutkin, Vitaly; Redenski, Idan; Steinberg, Doron

2015-03-01

The purpose of this study was to compare the efficacy of three irrigation techniques for smear-layer removal with 17% EDTA. Cleaning and shaping the root canal system during endodontic treatment produces a smear layer and hard tissue debris. Three irrigation techniques were tested for solution infiltration of this layer: positive-pressure irrigation, passive ultrasonic irrigation, and laser-activated irrigation. Sixty extracted teeth were divided into six equal groups; 17% EDTA was used for 60 sec irrigation of five of the groups. The groups were as follows: Group 1, treated only with ProTaper™ F3 Ni-Ti files; Group 2, positive-pressure irrigation, with a syringe; Group 3, passive ultrasonic irrigation, inserted 1 mm short of the working length; Group 4, passive ultrasonic irrigation, inserted in the upper coronal third of the root; Group 5, Er:YAG laser-activated irrigation, inserted 1 mm short of the working length; and Group 6, Er:YAG laser-activated irrigation, inserted in the upper coronal third of the root. Scanning electron microscopy showed that the smear layer is removed most efficiently using laser-activated irrigation at low energy with 17% EDTA, inserted either at the working length or only in the coronal upper third of the root. Amounts of Ca, P, and O were not significantly different on all treated dentin surfaces. Smear-layer removal was most effective when the root canals were irrigated using Er:YAG laser at low energy with 17% EDTA solution. Interestingly, removal of the smear layer along the entire canal was similar when the laser was inserted in the upper coronal third and at 1 mm short of the working length of the root canal. This effect was not observed with the ultrasonic and positive-pressure techniques.

Evaluation of two line probe assays for rapid detection of Mycobacterium tuberculosis, tuberculosis (TB) drug resistance, and non-TB Mycobacteria in HIV-infected individuals with suspected TB.

PubMed

Luetkemeyer, Anne F; Kendall, Michelle A; Wu, Xingye; Lourenço, Maria Cristina; Jentsch, Ute; Swindells, Susan; Qasba, Sarojini S; Sanchez, Jorge; Havlir, Diane V; Grinsztejn, Beatriz; Sanne, Ian M; Firnhaber, Cynthia

2014-04-01

Limited performance data from line probe assays (LPAs), nucleic acid tests used for the rapid diagnosis of tuberculosis (TB), nontuberculosis mycobacteria (NTM), and Mycobacterium tuberculosis drug resistance are available for HIV-infected individuals, in whom paucibacillary TB is common. In this study, the strategy of testing sputum with GenoType MTBDRplus (MTBDR-Plus) and GenoType Direct LPA (Direct LPA) was compared to a gold standard of one mycobacterial growth indicator tube (MGIT) liquid culture. HIV-positive (HIV(+)) individuals with suspected TB from southern Africa and South America with <7 days of TB treatment had 1 sputum specimen tested with Direct LPA, MTBDR-Plus LPA, smear microscopy, MGIT, biochemical identification of mycobacterial species, and culture-based drug-susceptibility testing (DST). Of 639 participants, 59.3% were MGIT M. tuberculosis culture positive, of which 276 (72.8%) were acid-fast bacillus (AFB) smear positive. MTBDR-Plus had a sensitivity of 81.0% and a specificity of 100%, with sensitivities of 44.1% in AFB smear-negative versus 94.6% in AFB smear-positive specimens. For specimens that were positive for M. tuberculosis by MTBDR-Plus, the sensitivity and specificity for rifampin resistance were 91.7% and 96.6%, respectively, and for isoniazid (INH) they were 70.6% and 99.1%. The Direct LPA had a sensitivity of 88.4% and a specificity of 94.6% for M. tuberculosis detection, with a sensitivity of 72.5% in smear-negative specimens. Ten of 639 MGIT cultures grew Mycobacterium avium complex or Mycobacterium kansasii, half of which were detected by Direct LPA. Both LPA assays performed well in specimens from HIV-infected individuals, including in AFB smear-negative specimens, with 72.5% sensitivity for M. tuberculosis identification with the Direct LPA and 44.1% sensitivity with MTBDR-Plus. LPAs have a continued role for use in settings where rapid identification of INH resistance and clinically relevant NTM are priorities.

Smear layer removal and canal cleanliness using different irrigation systems (EndoActivator, EndoVac, and passive ultrasonic irrigation): field emission scanning electron microscopic evaluation in an in vitro study.

PubMed

Mancini, Manuele; Cerroni, Loredana; Iorio, Lorenzo; Armellin, Emiliano; Conte, Gabriele; Cianconi, Luigi

2013-11-01

The purpose of this study was to evaluate the effectiveness of different irrigating methods in removing the smear layer at 1, 3, 5, and 8 mm from the apex of endodontic canals. Sixty-five extracted single-rooted human mandibular premolars were decoronated to a standardized length of 16 mm. Specimens were shaped to ProTaper F4 (Dentsply Maillefer, Ballaigues, Switzerland) and irrigated with 5.25% NaOCl at 37°C. Teeth were divided into 5 groups (2 control groups [n = 10] and 3 test groups [n = 15]) according to the final irrigant activation/delivering technique (ie, sonic irrigation, passive ultrasonic irrigation [PUI], or apical negative pressure). Root canals were then split longitudinally and observed by field emission scanning electron microscopy. The presence of debris and a smear layer at 1, 3, 5, and 8 mm from the apex was evaluated. Scores were analyzed by Kruskal-Wallis and Mann-Whitney U tests. The EndoActivator System (Dentsply Tulsa Dental Specialties, Tulsa, OK) was significantly more efficient than PUI and the control groups in removing the smear layer at 3, 5, and 8 mm from the apex. The EndoVac System (Discus Dental, Culver City, CA) removed statistically significantly more smear layer than all groups at 1, 3, 5, and 8 mm from the apex. At 5 and 8 mm from the apex, PUI and the EndoVac did not differ statistically significantly, but both performed statistically better than the control groups. In our study, none of the activation/delivery systems completely removed the smear layer from the endodontic dentine walls; nevertheless, the EndoActivator and EndoVac showed the best results at 3, 5, and 8 mm (EndoActivator) and 1, 3, 5, and 8 mm (EndoVac) from the apex. Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Utility of PCR in diagnosing pulmonary tuberculosis.

PubMed

Bennedsen, J; Thomsen, V O; Pfyffer, G E; Funke, G; Feldmann, K; Beneke, A; Jenkins, P A; Hegginbothom, M; Fahr, A; Hengstler, M; Cleator, G; Klapper, P; Wilkins, E G

1996-06-01

At present, the rapid diagnosis of pulmonary tuberculosis rests with microscopy. However, this technique is insensitive and many cases of pulmonary tuberculosis cannot be initially confirmed. Nucleic acid amplification techniques are extremely sensitive, but when they are applied to tuberculosis diagnosis, they have given variable results. Investigators at six centers in Europe compared a standardized PCR system (Amplicor; Roche) against conventional culture methods. Defined clinical information was collected. Discrepant samples were retested, and inhibition assays and backup amplification with a separate primer pair were performed. Mycobacterium tuberculosis complex organisms were recovered from 654 (9.1%) of 7,194 samples and 293 (7.8%) of 3,738 patients. Four hundred fifty-two of the M. tuberculosis isolates from 204 patients were smear positive and culture positive. Among the culture-positive specimens, PCR had a sensitivity of 91.4% for smear-positive specimens and 60.9% for smear-negative specimens, with a specificity of 96.1%. Analysis of 254 PCR-positive, culture-negative specimens with discrepant results revealed that 130 were from patients with recently diagnosed tuberculosis and 94 represented a presumed laboratory error. Similar analysis of 118 PCR-negative, culture-positive specimens demonstrated that 27 discrepancies were due to presumed uneven aliquot distribution and 11 were due to presumed laboratory error; PCR inhibitors were detected in 8 specimens. Amplicor enables laboratories with little previous experience with nucleic acid amplification to perform PCR. Disease in more than 60% of the patients with tuberculosis with smear-negative, culture-positive specimens can be diagnosed at the time of admission, and potentially all patients with smear-positive specimens can immediately be confirmed as being infected with M. tuberculosis, leading to improved clinical management.

Fluorescent microscopy and Ziehl-Neelsen staining of bronchoalveolar lavage, bronchial washings, bronchoscopic brushing and post bronchoscopic sputum along with cytological examination in cases of suspected tuberculosis.

PubMed

Bodal, Vijay Kumar; Bal, Manjit S; Bhagat, Sunita; Kishan, Jai; Brar, Rupinder K

2015-01-01

Ever since the discovery of Mycobacterium tuberculosis in 1882, many diagnostic methods have been developed. However "The gold standard" for the diagnosis of tuberculosis (TB) is still the demonstration of acid fast Bacilli (AFB) by microscopic examination of smear or bacteriological confirmation by culture method. In suspected 75 patients with active pulmonary TB, the materials obtained bronchoscopically, were bronchoalveolar lavage (BAL), bronchial brushings, bronchial washings and post bronchoscopic sputum. Four smears were made from each of the specimen. Fluorescent Staining, Ziehl-Neelsen (ZN), Pap and May Grunwald-Giemsa (MGG) stains were carried out for cytological examination. Fluorescent stain yielded maximum AFB positivity in all the methods, that is 36 (48%) in post fibre-optic bronchoscopy (FOB) sputum and 19 (25.33%) by fluorescence microscopy in both bronchial brushings and bronchial washings. Maximum yield of AFB with ZN staining 12 (16%) was equal to the post FOB sputum and bronchial brushings samples. It was followed by 6 cases (8%) in BAL and 4 (5.3%) in bronchial washings. The cytological examination was suggestive of TB in only 8 (10.66%) cases in bronchial washings and 6 (8%) cases in post FOB collection. It was equal in BAL and Bronchial brushings each that is 5 (6.67%). Bronchoscopy is a useful diagnostic tool and fluorescent microscopy is more sensitive than ZN and cytology. On X-ray examination, other diseases like malignancy or fungus can also mimick TB. So apart from ZN staining or fluorescence microscopy, Pap and MGG stain will be worthwhile to identify other microorganisms.

Diagnostic efficacy of microscopy, rapid diagnostic test and polymerase chain reaction for malaria using bayesian latent class analysis.

PubMed

Saha, Sreemanti; Narang, Rahul; Deshmukh, Pradeep; Pote, Kiran; Anvikar, Anup; Narang, Pratibha

2017-01-01

The diagnostic techniques for malaria are undergoing a change depending on the availability of newer diagnostics and annual parasite index of infection in a particular area. At the country level, guidelines are available for selection of diagnostic tests; however, at the local level, this decision is made based on malaria situation in the area. The tests are evaluated against the gold standard, and if that standard has limitations, it becomes difficult to compare other available tests. Bayesian latent class analysis computes its internal standard rather than using the conventional gold standard and helps comparison of various tests including the conventional gold standard. In a cross-sectional study conducted in a tertiary care hospital setting, we have evaluated smear microscopy, rapid diagnostic test (RDT), and polymerase chain reaction (PCR) for diagnosis of malaria using Bayesian latent class analysis. We found the magnitude of malaria to be 17.7% (95% confidence interval: 12.5%-23.9%) among the study subjects. In the present study, the sensitivity of microscopy was 63%, but it had very high specificity (99.4%). Sensitivity and specificity of RDT and PCR were high with RDT having a marginally higher sensitivity (94% vs. 90%) and specificity (99% vs. 95%). On comparison of likelihood ratios (LRs), RDT had the highest LR for positive test result (175) and the lowest LR for negative test result (0.058) among the three tests. In settings like ours conventional smear microscopy may be replaced with RDT and as we move toward elimination and facilities become available PCR may be roped into detect cases with lower parasitaemia.

Are current case-finding methods under-diagnosing tuberculosis among women in Myanmar? An analysis of operational data from Yangon and the nationwide prevalence survey.

PubMed

Khan, M S; Khine, T M; Hutchison, C; Coker, R J; Hane, K M; Innes, A L; Aung, S

2016-03-03

Although there is a large increase in investment for tuberculosis control in Myanmar, there are few operational analyses to inform policies. Only 34% of nationally reported cases are from women. In this study, we investigate sex differences in tuberculosis diagnoses in Myanmar in order to identify potential health systems barriers that may be driving lower tuberculosis case finding among women. From October 2014 to March 2015, we systematically collected data on all new adult smear positive tuberculosis cases in ten township health centres across Yangon, the largest city in Myanmar, to produce an electronic tuberculosis database. We conducted a descriptive cross-sectional analysis of sex differences in tuberculosis diagnoses at the township health centres. We also analysed national prevalence survey data to calculate additional case finding in men and women by using sputum culture when smear microscopy was negative, and estimated the sex-specific impact of using a more sensitive diagnostic tool at township health centres. Overall, only 514 (30%) out of 1371 new smear positive tuberculosis patients diagnosed at the township health centres were female. The proportion of female patients varied by township (from 21% to 37%, p = 0.0172), month of diagnosis (37% in February 2015 and 23% in March 2015 p = 0.0004) and age group (26% in 25-64 years and 49% in 18-25 years, p < 0.0001). Smear microscopy grading of sputum specimens was not substantially different between sexes. The prevalence survey analysis indicated that the use of a more sensitive diagnostic tool could result in the proportion of females diagnosed at township health centres increasing to 36% from 30%. Our study, which is the first to systematically compile and analyse routine operational data from tuberculosis diagnostic centres in Myanmar, found that substantially fewer women than men were diagnosed in all study townships. The sex ratio of newly diagnosed cases varied by age group, month of diagnosis and township of diagnosis. Low sensitivity of tuberculosis diagnosis may lead to a potential under-diagnosis of tuberculosis among women.

Comparison among four commonly used demineralizing agents for root conditioning. A scanning electron microscopy

PubMed Central

do AMARAL, Nathalia Godoy; de REZENDE, Maria Lúcia Rubo; HIRATA, Fabiana; RODRIGUES, Marcus Gustavo Silva; SANT'ANA, Adriana Campos Passanezi; GREGHI, Sebastião Luiz Aguiar; PASSANEZI, Euloir

2011-01-01

Dental roots that have been exposed to the oral cavity and periodontal pocket environment present superficial changes, which can prevent connective tissue reattachment. Demineralizing agents have been used as an adjunct to the periodontal treatment aiming at restoring the biocompatibility of roots. Objective This study compared four commonly used demineralizing agents for their capacity of removing smear layer and opening dentin tubules. Methods Fifty fragments of human dental roots previously exposed to periodontal disease were scaled and randomly divided into the following groups of treatment: 1) CA: demineralization with citric acid for 3 min; 2) TC-HCl: demineralization with tetracycline-HCl for 3 min; 3) EDTA: demineralization with EDTA for 3 min; 4) PA: demineralization with 37% phosphoric acid for 3 min; 5)Control: rubbing of saline solution for 3 min. Scanning electron microscopy was used to check for the presence of residual smear layer and for measuring the number and area of exposed dentin tubules. Results Smear layer was present in 100% of the specimens from the groups PA and control; in 80% from EDTA group; in 33.3% from TC-HCl group and 0% from CA group. The mean numbers of exposed dentin tubules in a standardized area were: TC-HCl=43.8±25.2; CA=39.3±37; PA=12.1±16.3; EDTA=4.4±7.5 and Control=2.3±5.7. The comparison showed significant differences between the following pairs of groups: TC-HCl and Control; TC-HCl and EDTA; CA and Control; and CA and EDTA. The mean percentages of area occupied by exposed dentin tubules were: CA=0.12±0.17%; TC-HCl=0.08±0.06%; PA=0.03±0.05%; EDTA=0.01±0.01% and Control=0±0%. The CA group differed significantly from the others except for the TC-HCl group. Conclusion There was a decreasing ability for smear layer removal and dentin tubule widening as follows: AC>TC-HCl>PA>EDTA. This information can be of value as an extra parameter for choosing one of them for root conditioning. PMID:21986651

Application of principal component analysis to multispectral-multimodal optical image analysis for malaria diagnostics.

PubMed

Omucheni, Dickson L; Kaduki, Kenneth A; Bulimo, Wallace D; Angeyo, Hudson K

2014-12-11

Multispectral imaging microscopy is a novel microscopic technique that integrates spectroscopy with optical imaging to record both spectral and spatial information of a specimen. This enables acquisition of a large and more informative dataset than is achievable in conventional optical microscopy. However, such data are characterized by high signal correlation and are difficult to interpret using univariate data analysis techniques. In this work, the development and application of a novel method which uses principal component analysis (PCA) in the processing of spectral images obtained from a simple multispectral-multimodal imaging microscope to detect Plasmodium parasites in unstained thin blood smear for malaria diagnostics is reported. The optical microscope used in this work has been modified by replacing the broadband light source (tungsten halogen lamp) with a set of light emitting diodes (LEDs) emitting thirteen different wavelengths of monochromatic light in the UV-vis-NIR range. The LEDs are activated sequentially to illuminate same spot of the unstained thin blood smears on glass slides, and grey level images are recorded at each wavelength. PCA was used to perform data dimensionality reduction and to enhance score images for visualization as well as for feature extraction through clusters in score space. Using this approach, haemozoin was uniquely distinguished from haemoglobin in unstained thin blood smears on glass slides and the 590-700 spectral range identified as an important band for optical imaging of haemozoin as a biomarker for malaria diagnosis. This work is of great significance in reducing the time spent on staining malaria specimens and thus drastically reducing diagnosis time duration. The approach has the potential of replacing a trained human eye with a trained computerized vision system for malaria parasite blood screening.

Optimizing morphology through blood cell image analysis.

PubMed

Merino, A; Puigví, L; Boldú, L; Alférez, S; Rodellar, J

2018-05-01

Morphological review of the peripheral blood smear is still a crucial diagnostic aid as it provides relevant information related to the diagnosis and is important for selection of additional techniques. Nevertheless, the distinctive cytological characteristics of the blood cells are subjective and influenced by the reviewer's interpretation and, because of that, translating subjective morphological examination into objective parameters is a challenge. The use of digital microscopy systems has been extended in the clinical laboratories. As automatic analyzers have some limitations for abnormal or neoplastic cell detection, it is interesting to identify quantitative features through digital image analysis for morphological characteristics of different cells. Three main classes of features are used as follows: geometric, color, and texture. Geometric parameters (nucleus/cytoplasmic ratio, cellular area, nucleus perimeter, cytoplasmic profile, RBC proximity, and others) are familiar to pathologists, as they are related to the visual cell patterns. Different color spaces can be used to investigate the rich amount of information that color may offer to describe abnormal lymphoid or blast cells. Texture is related to spatial patterns of color or intensities, which can be visually detected and quantitatively represented using statistical tools. This study reviews current and new quantitative features, which can contribute to optimize morphology through blood cell digital image processing techniques. © 2018 John Wiley & Sons Ltd.

Natural and synthetic oviposition stimulants forCatolaccus grandis (Burks) females.

PubMed

Guerra, A A; Martinez, S; Sonia Del Rio, H

1994-07-01

Oviposition behavior was elicited fromCatolaccus grandis (Burks) (Hymenoptera: Pteromalidae) females, an ectoparasitoid of the boll weevil,Anthonomus grandis Boheman (Coleoptera: Curculionidae), by smears of freshly cut cotton bolls or smears of extracts prepared with boll weevil damaged or undamaged cotton boll tissues. Oviposition behavior was also elicited fromC. grandis females by smears made withn-pentane,n-hexane,n-heptane, and isooctane. This is the first report of oviposition behavior elicited for any parasitoid by these short-chain saturated hydrocarbons (alkanes), introducing a new concept on the chemical mediation of parasitoid behavior during host selection. Oviposition behavior was also elicited fromC. grandis females by volatiles emanating from an artificial diet devoid of insect components that was specifically developed for the in vitro rearing of ectoparasitoids. The possible use of a synergistic combination ofn-hexane and diet to optimize the mechanized production of noncontaminated eggs is also discussed.

Feasibility, accuracy, and clinical effect of point-of-care Xpert MTB/RIF testing for tuberculosis in primary-care settings in Africa: a multicentre, randomised, controlled trial.

PubMed

Theron, Grant; Zijenah, Lynn; Chanda, Duncan; Clowes, Petra; Rachow, Andrea; Lesosky, Maia; Bara, Wilbert; Mungofa, Stanley; Pai, Madhukar; Hoelscher, Michael; Dowdy, David; Pym, Alex; Mwaba, Peter; Mason, Peter; Peter, Jonny; Dheda, Keertan

2014-02-01

The Xpert MTB/RIF test for tuberculosis is being rolled out in many countries, but evidence is lacking regarding its implementation outside laboratories, ability to inform same-day treatment decisions at the point of care, and clinical effect on tuberculosis-related morbidity. We aimed to assess the feasibility, accuracy, and clinical effect of point-of-care Xpert MTB/RIF testing at primary-care health-care facilities in southern Africa. In this pragmatic, randomised, parallel-group, multicentre trial, we recruited adults with symptoms suggestive of active tuberculosis from five primary-care health-care facilities in South Africa, Zimbabwe, Zambia, and Tanzania. Eligible patients were randomly assigned using pregenerated tables to nurse-performed Xpert MTB/RIF at the clinic or sputum smear microscopy. Participants with a negative test result were empirically managed according to local WHO-compliant guidelines. Our primary outcome was tuberculosis-related morbidity (measured with the TBscore and Karnofsky performance score [KPS]) in culture-positive patients who had begun anti-tuberculosis treatment, measured at 2 months and 6 months after randomisation, analysed by intention to treat. This trial is registered with Clinicaltrials.gov, number NCT01554384. Between April 12, 2011, and March 30, 2012, we randomly assigned 758 patients to smear microscopy (182 culture positive) and 744 to Xpert MTB/RIF (185 culture positive). Median TBscore in culture-positive patients did not differ between groups at 2 months (2 [IQR 0-3] in the smear microscopy group vs 2 [0·25-3] in the MTB/RIF group; p=0·85) or 6 months (1 [0-3] vs 1 [0-3]; p=0·35), nor did median KPS at 2 months (80 [70-90] vs 90 [80-90]; p=0·23) or 6 months (100 [90-100] vs 100 [90-100]; p=0·85). Point-of-care MTB/RIF had higher sensitivity than microscopy (154 [83%] of 185 vs 91 [50%] of 182; p=0·0001) but similar specificity (517 [95%] 544 vs 540 [96%] of 560; p=0·25), and had similar sensitivity to laboratory-based MTB/RIF (292 [83%] of 351; p=0·99) but higher specificity (952 [92%] of 1037; p=0·0173). 34 (5%) of 744 tests with point-of-care MTB/RIF and 82 (6%) of 1411 with laboratory-based MTB/RIF failed (p=0·22). Compared with the microscopy group, more patients in the MTB/RIF group had a same-day diagnosis (178 [24%] of 744 vs 99 [13%] of 758; p<0·0001) and same-day treatment initiation (168 [23%] of 744 vs 115 [15%] of 758; p=0·0002). Although, by end of the study, more culture-positive patients in the MTB/RIF group were on treatment due to reduced dropout (15 [8%] of 185 in the MTB/RIF group did not receive treatment vs 28 [15%] of 182 in the microscopy group; p=0·0302), the proportions of all patients on treatment in each group by day 56 were similar (320 [43%] of 744 in the MTB/RIF group vs 317 [42%] of 758 in the microscopy group; p=0·6408). Xpert MTB/RIF can be accurately administered by a nurse in primary-care clinics, resulting in more patients starting same-day treatment, more culture-positive patients starting therapy, and a shorter time to treatment. However, the benefits did not translate into lower tuberculosis-related morbidity, partly because of high levels of empirical-evidence-based treatment in smear-negative patients. European and Developing Countries Clinical Trials Partnership, National Research Foundation, and Claude Leon Foundation. Copyright © 2014 Elsevier Ltd. All rights reserved.

[Efficacy of a rapid test to diagnose Plasmodium vivax in symptomatic patients of Chiapas, Mexico].

PubMed

González-Cerón, Lilia; Rodríguez, Mario H; Betanzos, Angel F; Abadía, Acatl

2005-01-01

To evaluate, under laboratory conditions, the sensitivity and specificity of a rapid diagnostic test (OptiMAL), based on immunoreactive strips, to detect Plasmodium vivax infection in febrile patients in Southern Chiapas, Mexico. The presence of parasites in blood samples of 893 patients was investigated by Giemsa-stained thick blood smear microscopic examination (gold standard). A blood drop from the same sample was smeared on immunoreactive strips to investigate the presence of the parasite pLDH. Discordant results were resolved by PCR amplification of the parasite's 18S SSU rRNA, to discard infection. OptiMAL had an overall sensitivity of 93.3% and its specificity was 99.5%. Its positive and negative predictive values were 96.5% and 98.9%, respectively. Signal intensity in OptiMAL strips correlated well with the parasitemia density in the blood samples (r = 0.601, p = 0.0001). This rapid test had acceptable sensitivity and specificity to detect P. vivax under laboratory conditions and could be useful for malaria diagnosis in field operations in Mexico.

The External Quality Assessment Scheme (EQAS): Experiences of a medium sized accredited laboratory.

PubMed

Bhat, Vivek; Chavan, Preeti; Naresh, Chital; Poladia, Pratik

2015-06-15

We put forth our experiences of EQAS, analyzed the result discrepancies, reviewed the corrective actions and also put forth strategies for risk identification and prevention of potential errors in a medical laboratory. For hematology, EQAS samples - blood, peripheral and reticulocyte smears - were received quarterly every year. All the blood samples were processed on HMX hematology analyzer by Beckman-Coulter. For clinical chemistry, lyophilized samples were received and were processed on Siemens Dimension Xpand and RXL analyzers. For microbiology, EQAS samples were received quarterly every year as lyophilized strains along with smears and serological samples. In hematology no outliers were noted for reticulocyte and peripheral smear examination. Only one outlier was noted for CBC. In clinical chemistry outliers (SDI ≥ 2) were noted in 7 samples (23 parameters) out of total 36 samples (756 parameters) processed. Thirteen of these parameters were analyzed as random errors, 3 as transcriptional errors and seven instances of systemic error were noted. In microbiology, one discrepancy was noted in isolate identification and in the grading of smears for AFB by Ziehl Neelsen stain. EQAS along with IQC is a very important tool for maintaining optimal quality of services. Copyright © 2015 Elsevier B.V. All rights reserved.

Xpert® Mtb/Rif assay for pulmonary tuberculosis and rifampicin resistance in adults

PubMed Central

Steingart, Karen R; Sohn, Hojoon; Schiller, Ian; Kloda, Lorie A; Boehme, Catharina C; Pai, Madhukar; Dendukuri, Nandini

2013-01-01

Background Accurate and rapid detection of tuberculosis (TB) and drug resistance are critical for improving patient care and decreasing the spread of TB. Xpert® MTB/RIF assay (Xpert) is a rapid, automated test that can detect both TB and rifampicin resistance, within two hours after starting the test, with minimal hands-on technical time, but is more expensive than conventional sputum microscopy. Objectives To assess the diagnostic accuracy of Xpert for pulmonary TB (TB detection), both where Xpert was used as an initial test replacing microscopy, and where Xpert was used as an add-on test following a negative smear microscopy result. To assess the diagnostic accuracy of Xpert for rifampicin resistance detection where Xpert was used as the initial test, replacing conventional culture-based drug susceptibility testing. The population of interest was adults suspected of having pulmonary TB or multidrug-resistant TB (MDR-TB), with or without HIV infection. Search methods We performed a comprehensive search of the following databases: Cochrane Infectious Diseases Group Specialized Register; MEDLINE; EMBASE; ISI Web of Knowledge; MEDION; LILACS; BIOSIS; and SCOPUS. We also searched the metaRegister of Controlled Trials (mRCT) and the search portal of the WHO International Clinical Trials Registry Platform to identify ongoing trials. We performed searches on 25 September 2011 and we repeated them on 15 December 2011, without language restriction. Selection criteria We included randomized controlled trials, cross-sectional, and cohort studies that used respiratory specimens to compare Xpert with culture for detecting TB and Xpert with conventional phenotypic drug susceptibility testing for detecting rifampicin resistance. Data collection and analysis For each study, two review authors independently extracted a set of data using a standardized data extraction form. When possible, we extracted data for subgroups by smear and HIV status. We assessed the quality of studies using the QUADAS-2 tool. We carried out meta-analyses to estimate the pooled sensitivity and specificity of Xpert separately for TB detection and rifampicin resistance detection using a bivariate random-effects model. We estimated the median pooled sensitivity and specificity and their 95% credible intervals (CrI). Main results We identified 18 unique studies as eligible for this review, including two multicentre international studies, one with five and the other with six distinct study centres. The majority of studies (55.6%) were performed in low-income and middle-income countries. In 17 of the 18 studies, Xpert was performed by trained technicians in reference laboratories. When used as an initial test replacing smear microscopy (15 studies, 7517 participants), Xpert achieved a pooled sensitivity of 88% (95% CrI 83% to 92%) and pooled specificity of 98% (95% CrI 97% to 99%). As an add-on test following a negative smear microscopy result (14 studies, 5719 participants), Xpert yielded a pooled sensitivity of 67% (95% CrI 58% to 74%) and pooled specificity of 98% (95% CrI 97% to 99%). In clinical subgroups, we found the following accuracy estimates: the pooled sensitivity was 98% (95% CrI 97% to 99%) for smear-positive, culture-positive TB and 68% (95% CrI 59% to 75%) for smear-negative, culture-positive TB (15 studies); the pooled sensitivity was 80% (95% CrI 67% to 88%) in people living with HIV and 89% (95% CrI 81% to 94%) in people without HIV infection (four studies). For rifampicin resistance detection (11 studies, 2340 participants), Xpert achieved a pooled sensitivity of 94% (95% CrI 87% to 97%) and pooled specificity of 98% (95% CrI 97% to 99%). In a separate analysis, Xpert could distinguish between TB and nontuberculous mycobacteria (NTM) in clinical samples with high accuracy: among 139 specimens with NTM, Xpert was positive in only one specimen that grew NTM. In a hypothetical cohort of 1000 individuals suspected of having rifampicin resistance (a proxy for MDR-TB), where the prevalence of rifampicin resistance is 30%, we estimated that on average Xpert would wrongly identify 14 patients as being rifampicin resistant. In comparison, where the prevalence of rifampicin resistance is only 2%, we estimated that the number of individuals wrongly identified as rifampicin resistant would increase to 20, an increase of 43%. Authors' conclusions This review shows that Xpert used as an initial diagnostic test for TB detection and rifampicin resistance detection in patients suspected of having TB, MDR-TB, or HIV-associated TB is sensitive and specific. Xpert may also be valuable as an add-on test following microscopy for patients who have previously been found to be smear-negative. An Xpert result that is positive for rifampicin resistance should be carefully interpreted and take into consideration the risk of MDR-TB in a given patient and the expected prevalence of MDR-TB in a given setting. Studies in this review mainly assessed sensitivity and specificity of the test when used in reference laboratories in research investigations. Most studies were performed in high TB burden countries. Ongoing use of Xpert in high TB burden countries will contribute to the evidence base on the diagnostic accuracy and clinical impact of Xpert in routine programmatic and peripheral health care settings, including settings where the test is performed at the point of care. PMID:23440842

Partial Characterization of a Novel Amphibian Hemoglobin as a Model for Graduate Student Investigation on Peptide Chemistry, Mass Spectrometry, and Atomic Force Microscopy

ERIC Educational Resources Information Center

Bemquerer, Marcelo P.; Macedo, Jessica K. A.; Ribeiro, Ana Carolina J.; Carvalho, Andrea C.; Silva, Debora O. C.; Braz, Juliana M.; Medeiros, Kelliane A.; Sallet, Lunalva A. P.; Campos, Pollyanna F.; Prates, Maura V.; Silva, Luciano P.

2012-01-01

Graduate students in chemistry, and in biological and biomedical fields must learn the fundamentals and practices of peptide and protein chemistry as early as possible. A project-oriented approach was conducted by first-year M.Sc and Ph.D students in biological sciences. A blind glass slide containing a cellular smear and an aqueous cellular…

Market assessment of tuberculosis diagnostics in China in 2012.

PubMed

Zhao, Y-L; Pang, Y; Xia, H; Du, X; Chin, D; Huan, S-T; Dong, H-Y; Zhang, Z-Y; Ginnard, J; Perkins, M D; Boehme, C C; Jefferson, C; Pantoja, A; Qin, Z Z; Chedore, P; Denkinger, C M; Pai, M; Kik, S V

2016-03-01

To assess the 2012 served available market for tuberculosis (TB) diagnostics in China in the sector served by the China Centre for Disease Control and Prevention (CDC) and the hospital sector in China, including both designated TB hospitals and general hospitals. Test volumes and unit costs were assessed for tuberculin skin tests, interferon-gamma release assays (IGRAs), smear microscopy, serology, cultures, speciation tests, nucleic-acid amplification tests (NAATs), drug susceptibility tests and adenosine-deaminase tests (ADA). Data were obtained from electronic databases (CDC sector) and through surveys (hospital sector), and were estimated for the two sectors and for the country as a whole. Test costs were estimated by staff at China CDC, and using published literature. In 2012, the China CDC and hospital sectors performed a total of 44 million TB diagnostic tests at an overall value of US$294 million. Tests used by the CDC sector were smear microscopy, solid and liquid culture and DST, while the hospital sector also used IGRAs, NAATs, ADA and serology. The hospital sector accounted for 76% of the overall test volume and 94% of the market value. China has a very large TB diagnostic market that encompasses a wide range of diagnostic tests, with the majority being performed in Chinese hospitals.

Market assessment of tuberculosis diagnostics in India in 2013.

PubMed

Maheshwari, P; Chauhan, K; Kadam, R; Pujani, A; Kaur, M; Chitalia, M; Dabas, H; Perkins, M D; Boehme, C C; Denkinger, C M; Raizada, N; Ginnard, J; Jefferson, C; Pantoja, A; Rupert, S; Kik, S V; Cohen, C; Chedore, P; Satyanarayana, S; Pai, M

2016-03-01

India represents a significant potential market for new tests. We assessed India's market for tuberculosis (TB) diagnostics in 2013. Test volumes and unit costs were assessed for tuberculin tests, interferon-gamma release assays, sputum smear microscopy, serology, culture, speciation testing, nucleic-acid amplification tests (i.e., in-house polymerase chain reaction, Xpert(®) MTB/RIF, line-probe assays) and drug susceptibility testing. Data from the public sector were collected from the Revised National TB Control Programme reports. Private sector data were collected through a survey of private laboratories and practitioners. Data were also collected from manufacturers. In 2013, India's public sector performed 19.2 million tests, with a market value of US$22.9 million. The private sector performed 13.6 million tests, with a market value of US$60.4 million when prices charged to the patient were applied. The overall market was US$70.8 million when unit costs from the ingredient approach were used for the 32.8 million TB tests performed in the entire country. Smear microscopy was the most common test performed, accounting for 25% of the overall market value. India's estimated market value for TB diagnostics in 2013 was US$70.8 million. These data should be of relevance to test developers, donors and implementers.

Liquid-based cytology for primary cervical cancer screening: a multi-centre study

PubMed Central

Monsonego, J; Autillo-Touati, A; Bergeron, C; Dachez, R; Liaras, J; Saurel, J; Zerat, L; Chatelain, P; Mottot, C

2001-01-01

The aim of this six-centre, split-sample study was to compare ThinPrep fluid-based cytology to the conventional Papanicolaou smear. Six cytopathology laboratories and 35 gynaecologists participated. 5428 patients met the inclusion criteria (age > 18 years old, intact cervix, informed consent). Each cervical sample was used first to prepare a conventional Pap smear, then the sampling device was rinsed into a PreservCyt vial, and a ThinPrep slide was made. Screening of slide pairs was blinded (n = 5428). All non-negative concordant cases (n = 101), all non-concordant cases (n = 206), and a 5% random sample of concordant negative cases (n = 272) underwent review by one independent pathologist then by the panel of 6 investigators. Initial (blinded) screening results for ThinPrep and conventional smears were correlated. Initial diagnoses were correlated with consensus cytological diagnoses. Differences in disease detection were evaluated using McNemar's test. On initial screening, 29% more ASCUS cases and 39% more low-grade squamous intraepithelial lesions (LSIL) and more severe lesions (LSIL+) were detected on the ThinPrep slides than on the conventional smears (P = 0.001), including 50% more LSIL and 18% more high-grade SIL (HSIL). The ASCUS:SIL ratio was lower for the ThinPrep method (115:132 = 0.87:1) than for the conventional smear method (89:94 = 0.95:1). The same trend was observed for the ASCUS/AGUS:LSIL ratio. Independent and consensus review confirmed 145 LSIL+ diagnoses; of these, 18% more had been detected initially on the ThinPrep slides than on the conventional smears (P = 0.041). The ThinPrep Pap Test is more accurate than the conventional Pap test and has the potential to optimize the effectiveness of primary cervical cancer screening. © 2001 Cancer Research Campaign http://www.bjcancer.com PMID:11161401

A Comparative SEM Investigation of Smear Layer Remaining on Dentinal Walls by Three Rotary NiTi Files with Different Cross Sectional Designs in Moderately Curved Canals

PubMed Central

Kakkar, Pooja; Vats, Asit

2015-01-01

Objective: The objective of this study was to compare the smear layer formed on root canal walls during canal preparation of extracted human teeth by Twisted, Mtwo, and ProTaper rotary nickel titanium instruments. Materials and Methods: Sixty single rooted human premolar teeth with root curvature <250 were selected and randomly divided into three Groups (n= 20 teeth per Group). Three types of rotary nickel titanium instruments were used, Twisted (SybronEndo, Orange, CA, USA), Mtwo (VDW, Munich, Germany) and ProTaper (Dentsply Maillefer, Ballaigues, Switzerland) according to manufacturer’s instructions to instrument the root canals. Irrigation for all groups was performed after each instrument change with 3ml of 3% sodium hypochlorite followed by Glyde (File Prep, Dentsply, Maillefer, Ballaigues, Switzerland) as chelator paste and lubricant. Three different areas (coronal, middle and apical thirds) of the root canal were evaluated using scanning electron microscopy (SEM). The canal wall of each sample was assessed and compared using a predefined scale for the presence or absence of smear layer. Data were analysed statistically using ANOVA and Tukey HSD test Results: All three groups showed statistically significant more smear layer in the apical thirds of the canal as compared to the coronal and middle thirds (p<0.001). Mtwo rotary file system produced significantly less smear layer (p<0.001) compared to Twisted and ProTaper rotary instruments in the apical portion. Twisted Files resulted in less smear layer formation in the apical thirds of the canal compared to ProTaper rotary instruments but were statistically insignificant. Conclusion: Completely clean root canals were not found after instrumentation with any of the three instruments. Under the confines of this study Mtwo instruments produced significantly cleaner dentin wall surfaces throughout the canal length in comparison to Twisted and ProTaper rotary files. Twisted Files proved to be comparable to ProTaper rotary instruments with respect to canal cleanliness in the apical thirds of the root canal. PMID:25954704

Fluorescence Spectrum and Decay Measurement for Hsil VS Normal Cytology Differentiation in Liquid Pap Smear Supernatant

NASA Astrophysics Data System (ADS)

Vaitkuviene, A.; Gegzna, V.; Juodkazis, S.; Jursenas, S.; Miasojedovas, S.; Kurtinaitiene, R.; Rimiene, J.; Vaitkus, J.

2009-06-01

Cervical smear material contains endo and exocervical cells, mucus and inflammative, immune cells in cases of pathology. Just not destroyed keratinocytes lay on the glass for microscopy. Liquid cytology supernatant apart other diagnostics could be used for photodiagnostic. The spectroscopic parameters suitable for Normal and HSIL cytology groups supernatant differentiation are demonstrated. The dried liquid PAP supernatant fractions—sediment and liquid were investigated. Excitation and emission matrices (EEM), supernatant fluorescence decay measured under 280 nm diode short pulse excitation and fluorescence spectroscopy by excitation with 355 nm laser light were analyzed. The differences between Normal and HSIL groups were statistically proven in the certain spectral regions. Fluorescence decay peculiarities show spectral regions consisting of few fluorophores. Obtained results on fluorescence differences in Normal and HSIL groups' supernatant shows the potency of photodiagnosis application in cervical screening.

[Detection of Clonorchis sinensis eggs in the ground gallbladder stones by microscopy].

PubMed

Ma, Rui-Hong; Qiao, Tie; Luo, Xiao-Bing

2012-08-30

Sera, feces, bile and gallbladder stones were collected from 179 patients who accepted gallbladder-preserving cholelithotomy during the period of January to June 2010 at the general surgery department in the Second People's Hospital of Panyu District in Guangzhou. Rapid colloidal gold immunochromatography was used to detect IgG against Clonorchis sinensis. C. sinensis eggs were examined by fecal direct smear, and in bile sediments and ground gallbladder stones. The results showed that the positive rate of rapid colloidal gold immunochromatographic assay for IgG was 51.4%, and the egg positive rate in feces, bile sediments and gallbladder stones was 30.7%, 44.7% and 69.8%, respectively. The detection rate of fecal direct smear was the lowest, while that of the gallbladder stone examination was the highest (P < 0.05). Those patients with egg-positive feces and/or bile sediments were all with egg-positive gallbladder stones.

Evaluation of tartar control dentifrices in in vitro models of dentin sensitivity.

PubMed

Mason, S; Levan, A; Crawford, R; Fisher, S; Gaffar, A

1991-01-01

The effects of anticalculus dentifrices were compared with other commercially available dentifrices in in vitro models of dentin sensitivity. Changes in the hydraulic conductance of dentin discs were measured with and without a smear layer before and after treatment and also after a post-treatment acid etch. The capacity of dentifrices to occlude open dentinal tubules in vitro was also assessed by scanning electron microscopy (SEM). There was good correlation (R = 0.98) between our test and values reported in the literature. Tartar control dentifrices gave reductions in fluid flow rates through the dentin discs comparable to those obtained with Promise, Sensodyne, Thermodent and Denquel. Additionally, tartar control dentifrices did not remove microcrystalline debris (smear layers) from the surfaces of dentin in vitro. These results were confirmed by SEM. Thus, according to the hydrodynamic theory of dentin sensitivity, these in vitro results suggest that pyrophosphate-containing dentifrices should reduce dentinal sensitivity.

Diagnosing malaria in pregnancy: an update

PubMed Central

Fried, Michal; Muehlenbachs, Atis; Duffy, Patrick E

2013-01-01

Pregnancy malaria (PM) due to Plasmodium falciparum is a major cause of morbidity and mortality for women and their offspring, but is difficult to recognize and diagnose. During PM, parasites typically sequester in the placenta, whereas peripheral blood smears often appear negative. In addition, many infected women remain asymptomatic, especially in areas of high transmission where systemic immunity is high, although sequelae including maternal anemia and intrauterine growth retardation develop insidiously and increase mortality. New rapid diagnostic tests (RDTs) have shown promise for malaria diagnosis in nonpregnant individuals, including a product recently approved by the US FDA for use in the USA. However, the sensitivity and specificity of RDTs for diagnosis of PM may be suboptimal. Here, we review the methods that are used to detect or diagnose PM, including blood smear microscopy, RDTs, PCR-based methods, and finally placental histology, which is often cited as the gold standard for use in research studies and clinical trials. PMID:23199403

The counting of native blood cells by digital microscopy

NASA Astrophysics Data System (ADS)

Torbin, S. O.; Doubrovski, V. A.; Zabenkov, I. V.; Tsareva, O. E.

2017-03-01

An algorithm for photographic images processing of blood samples in its native state was developed to determine the concentration of erythrocytes, leukocytes and platelets without individual separate preparation of cells' samples. Special "photo templates" were suggested to use in order to identify red blood cells. The effect of "highlighting" of leukocytes, which was found by authors, was used to increase the accuracy of this type of cells counting. Finally to raise the resolution of platelets from leukocytes the areas of their photo images were used, but not their sizes. It is shown that the accuracy of cells counting for native blood samples may be comparable with the accuracy of similar studies for smears. At the same time the proposed native blood analysis simplifies greatly the procedure of sample preparation in comparison to smear, permits to move from the detection of blood cells ratio to the determination of their concentrations in the sample.

Correlation between automatic detection of malaria on thin film and experts' parasitaemia scores

NASA Astrophysics Data System (ADS)

Sunarko, Budi; Williams, Simon; Prescott, William R.; Byker, Scott M.; Bottema, Murk J.

2017-03-01

An algorithm was developed to diagnose the presence of malaria and to estimate the depth of infection by automatically counting individual normal and infected erythrocytes in images of thin blood smears. During the training stage, the parameters of the algorithm were optimized to maximize correlation with estimates of parasitaemia from expert human observers. The correlation was tested on a set of 1590 images from seven thin film blood smears. The correlation between the results from the algorithm and expert human readers was r = 0.836. Results indicate that reliable estimates of parasitaemia may be achieved by computational image analysis methods applied to images of thin film smears. Meanwhile, compared to biological experiments, the algorithm fitted well the three high parasitaemia slides and a mid-level parasitaemia slide, and overestimated the three low parasitaemia slides. To improve the parasitaemia estimation, the sources of the overestimation were identified. Emphasis is laid on the importance of further research in order to identify parasites independently of their erythrocyte hosts

Ultramorphology of the root surface subsequent to hand-ultrasonic simultaneous instrumentation during non-surgical periodontal treatments. An in vitro study

PubMed Central

D. ASPRIELLO, Simone; PIEMONTESE, Matteo; LEVRINI, Luca; SAURO, Salvatore

2011-01-01

Objective The purpose of this study was to investigate the ultramorphology of the root surfaces induced by mechanical instrumentation performed using conventional curettes or piezoelectric scalers when used single-handedly or with a combined technique. Material and Methods Thirty single-rooted teeth were selected and divided into 3 groups: Group A, instrumentation with curettes; Group B instrumentation with titanium nitride coated periodontal tip mounted in a piezoelectric handpiece; Group C, combined technique with curette/ultrasonic piezoelectric instrumentation. The specimens were processed and analyzed using confocal and scanning electron microscopy. Differences between the different groups of instrumentation were determined using Pearson’s χ 2 with significance predetermined at α=0.001. Results Periodontal scaling and root planing performed with curettes, ultrasonic or combined instrumentation induced several morphological changes on the root surface. The curettes produced a compact and thick multilayered smear layer, while the morphology of the root surfaces after ultrasonic scaler treatment appeared irregular with few grooves and a thin smear layer. The combination of curette/ultrasonic instrumentation showed exposed root dentin tubules with a surface morphology characterized by the presence of very few grooves and slender remnants of smear layer which only partially covered the root dentin. In some cases, it was also possible to observe areas with exposed collagen fibrils. Conclusion The curette-ultrasonic simultaneous instrumentation may combine the beneficial effects of each instrument in a single technique creating a root surface relatively free from the physical barrier of smear layer and dentin tubules orifices partial occlusion. PMID:21437474

Infection of apical dentin and root-end cavity disinfection.

PubMed

Aziz, Abdul; Chandler, Nicholas P; Hauman, Catharina H J; Leichter, Jonathan W; McNaughton, Andrew; Tompkins, Geoffrey R

2012-10-01

The purpose of this study was to assess Enterococcus faecalis penetration into the dentin of the apical 3 mm and bacterial death after the application of either chlorhexidine or laser to root-end cavities. Root canals of 60 single-rooted teeth were prepared. In part 1, cementum was removed semicircumferentially from 21 roots, and the smear layer was removed from 15 roots using 17% EDTA/cetrimide. Teeth were inoculated and incubated with E. faecalis for 10 days, rinsed, and live/dead stained. The effect of cementum and smear on bacterial penetration was assessed by confocal laser scanning microscopy (CLSM). In part 2, 39 teeth had root ends resected and cavities ultrasonically prepared. Inoculated roots were assigned to 1 of the following 3 groups: (1) root-end cavities irrigated with 0.2 % chlorhexidine, (2) root-end cavities irradiated with a laser for 20 seconds at 1.5 W, or (3) root-end cavities that received no treatment. Roots were live/dead stained, sectioned, and examined by CLSM. The depth of the bacterial penetration and bacterial survival were compared using the Mann-Whitney U test. The presence of a smear layer and/or cementum did not significantly affect bacterial penetration. In root-end cavities, chlorhexidine was more effective than laser (P < .001), reducing bacterial viability by 93% versus 70% with a laser. E. faecalis invaded the entire width of dentin in the apical 3 mm irrespective of the smear layer and/or cementum. Chlorhexidine was more effective than laser in disinfecting root-end cavities. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Bacterial vaginosis, aerobic vaginitis, vaginal inflammation and major Pap smear abnormalities.

PubMed

Vieira-Baptista, P; Lima-Silva, J; Pinto, C; Saldanha, C; Beires, J; Martinez-de-Oliveira, J; Donders, G

2016-04-01

The purpose of this investigation was to evaluate the impact of the vaginal milieu on the presence of abnormal Pap smears and a positive human papilloma virus (HPV) test. A cross-sectional study was conducted between June 2014 and May 2015, evaluating the vaginal discharge by fresh wet mount microscopy and comparing these data with Pap smear findings. Wet mount slides were scored for bacterial vaginosis (BV), aerobic vaginitis (AV), presence of Candida and Trichomonas vaginalis. Cytologic evaluation was done on all Pap smears according to the Bethesda criteria. The cobas© HPV Test (Roche) was performed for HPV detection. A total of 622 cases were evaluated. The mean age of the patients was 41.6 ± 10.65 years (range 21-75). Eighty-three women (13.3 %) had a cytology result worse than low-grade squamous intraepithelial lesion (LSIL). When comparing this group with the one with normal or minor [atypical squamous cells of undetermined significance (ASC-US) or LSIL] Pap smear abnormalities, there were no differences in the presence of Candida (32.5 % vs. 33.2 %, p = 1.0), absence of lactobacilli (38.6 % vs. 32.5 %, p = 0.32) or BV (20.5 % vs. 13.2 %, p = 0.09). On the other hand, moderate or severe inflammation (msI) (41.0 % vs. 28.8 %, p = 0,04), moderate or severe AV (msAV) (16.9 % vs. 7.2 %, p = 0.009) and msAV/BV (37.3 % vs. 20.0 %, p = 0.001) were more common in women with such major cervical abnormalities. No significant association was found between deviations of the vaginal milieu and high-risk HPV infection. The presence of msI or msAV, but not BV, is independently associated with an increased risk of major cervical cytological abnormalities, but not with HPV infection.

Pre-trained convolutional neural networks as feature extractors toward improved malaria parasite detection in thin blood smear images.

PubMed

Rajaraman, Sivaramakrishnan; Antani, Sameer K; Poostchi, Mahdieh; Silamut, Kamolrat; Hossain, Md A; Maude, Richard J; Jaeger, Stefan; Thoma, George R

2018-01-01

Malaria is a blood disease caused by the Plasmodium parasites transmitted through the bite of female Anopheles mosquito. Microscopists commonly examine thick and thin blood smears to diagnose disease and compute parasitemia. However, their accuracy depends on smear quality and expertise in classifying and counting parasitized and uninfected cells. Such an examination could be arduous for large-scale diagnoses resulting in poor quality. State-of-the-art image-analysis based computer-aided diagnosis (CADx) methods using machine learning (ML) techniques, applied to microscopic images of the smears using hand-engineered features demand expertise in analyzing morphological, textural, and positional variations of the region of interest (ROI). In contrast, Convolutional Neural Networks (CNN), a class of deep learning (DL) models promise highly scalable and superior results with end-to-end feature extraction and classification. Automated malaria screening using DL techniques could, therefore, serve as an effective diagnostic aid. In this study, we evaluate the performance of pre-trained CNN based DL models as feature extractors toward classifying parasitized and uninfected cells to aid in improved disease screening. We experimentally determine the optimal model layers for feature extraction from the underlying data. Statistical validation of the results demonstrates the use of pre-trained CNNs as a promising tool for feature extraction for this purpose.

The Effect of Fragaria vesca Extract on Smear Layer Removal: A Scanning Electron Microscopic Evaluation

PubMed Central

Davoudi, Amin; Razavi, Sayed Alireza; Mosaddeghmehrjardi, Mohammad Hossein; Tabrizizadeh, Mehdi

2015-01-01

Introduction: Successful endodontic treatment depends on elimination of the microorganisms through chemomechanical debridement. The aim of this in vitro study was to evaluate the effectiveness of Fragaria vesca (wild strawberry) extract (FVE) on the removal of smear layer (SL). Methods and Materials: In this analytical-observational study, 40 extracted mandibular and maxillary human teeth were selected. After canal preparation with standard step-back technique, the teeth were randomly divided into 4 groups according to the irrigation solution: saline (negative control), 5.25% NaOCl+EDTA (positive control), FVE and FVE+EDTA. The teeth were split longitudinally so that scanning electron microscopy (SEM) photomicrographs could be taken to evaluate the amount of remnant SL in coronal, middle and apical thirds. The data were analyzed statistically by the Kruskal-Wallis and Mann Whitney U tests and the level of significance was set at 0.05. Results: Significant differences were found among the groups (P<0.001). The use of NaOCl+EDTA was the most effective regimen for removing the SL followed by FVE+EDTA. FVE alone was significantly more effective than saline (P<0.001). Conclusion: FVE with and without EDTA could effectively remove the smear layer; however, compared to NaOCl group it was less effective. PMID:26526069

Diagnostic accuracy of a point-of-care urine test for tuberculosis screening among newly-diagnosed HIV-infected adults: a prospective, clinic-based study.

PubMed

Drain, Paul K; Losina, Elena; Coleman, Sharon M; Giddy, Janet; Ross, Douglas; Katz, Jeffrey N; Walensky, Rochelle P; Freedberg, Kenneth A; Bassett, Ingrid V

2014-02-26

A rapid diagnostic test for active tuberculosis (TB) at the clinical point-of-care could expedite case detection and accelerate TB treatment initiation. We assessed the diagnostic accuracy of a rapid urine lipoarabinomannan (LAM) test for TB screening among HIV-infected adults in a TB-endemic setting. We prospectively enrolled newly-diagnosed HIV-infected adults (≥18 years) at 4 outpatient clinics in Durban from Oct 2011-May 2012, excluding those on TB therapy. A physician evaluated all participants and offered CD4 cell count testing. Trained study nurses collected a sputum sample for acid-fast bacilli smear microscopy (AFB) and mycobacterial culture, and performed urine LAM testing using Determine™ TB LAM in the clinic. The presence of a band regardless of intensity on the urine LAM test was considered positive. We defined as the gold standard for active pulmonary TB a positive sputum culture for Mycobacterium tuberculosis. Diagnostic accuracy of urine LAM was assessed, alone and in combination with smear microscopy, and stratified by CD4 cell count. Among 342 newly-diagnosed HIV-infected participants, 190 (56%) were male, mean age was 35.6 years, and median CD4 was 182/mm3. Sixty participants had culture-positive pulmonary TB, resulting in an estimated prevalence of 17.5% (95% CI 13.7-22.0%). Forty-five (13.2%) participants were urine LAM positive. Mean time from urine specimen collection to LAM test result was 40 minutes (95% CI 34-46 minutes). Urine LAM test sensitivity was 28.3% (95% CI 17.5-41.4) overall, and 37.5% (95% CI 21.1-56.3) for those with CD4 count <100/mm3, while specificity was 90.1% (95% CI 86.0-93.3) overall, and 86.9% (95% CI 75.8-94.2) for those with CD4 < 100/mm3. When combined with sputum AFB (either test positive), sensitivity increased to 38.3% (95% CI 26.0-51.8), but specificity decreased to 85.8% (95% CI 81.1-89.7). In this prospective, clinic-based study with trained nurses, a rapid urine LAM test had low sensitivity for TB screening among newly-diagnosed HIV-infected adults, but improved sensitivity when combined with sputum smear microscopy.

Guidance for National Tuberculosis Programmes on the management of tuberculosis in children. Chapter 1: introduction and diagnosis of tuberculosis in children.

PubMed

2006-10-01

About one million children develop tuberculosis (TB) annually worldwide, accounting for about 11% of all TB cases. Children with TB differ from adults in their immunological and pathophysiological response in ways that may have important implications for the prevention, diagnosis and treatment of TB in children. There is an urgent need to improve the diagnosis and management of children with TB, and the prevention of TB in children, by ensuring their inclusion under the implementation of the Stop TB strategy by National TB Programmes. Critical areas for further research include a better understanding of the epidemiology of childhood TB, vaccine development, the development of better diagnostic techniques, new drug development, and the optimal formulations and dosing of first- and second-line TB drugs in children. Specifically regarding the diagnosis of TB in children, this relies on a careful and thorough assessment of all the evidence derived from a careful history, clinical examination and relevant investigations, e.g., tuberculin skin test, chest radiograph and sputum smear microscopy. Although bacteriological confirmation of TB is not always possible, it should be sought whenever possible, e.g., by sputum microscopy in children with suspected pulmonary TB who are old enough to produce a sputum sample. A trial of treatment with TB medications is not generally recommended as a method to diagnose TB in children. New, improved diagnostic tests are urgently needed.

Requirements for diagnosis of malaria at different levels of the laboratory network in Africa.

PubMed

Long, Earl G

2009-06-01

The rapid increase of resistance to cheap, reliable antimalarials, the increasing cost of effective drugs, and the low specificity of clinical diagnosis has increased the need for more reliable diagnostic methods for malaria. The most commonly used and most reliable remains microscopic examination of stained blood smears, but this technique requires skilled personnel, precision instruments, and ideally a source of electricity. Microscopy has the advantage of enabling the examiner to identify the species, stage, and density of an infection. An alternative to microscopy is the rapid diagnostic test (RDT), which uses a labeled monoclonal antibody to detect circulating parasitic antigens. This test is most commonly used to detect Plasmodium falciparum infections and is available in a plastic cassette format. Both microscopy and RDTs should be available at all levels of laboratory service in endemic areas, but in peripheral laboratories with minimally trained staff, the RDT may be a more practical diagnostic method.

Dietary supplementation with docosahexanoic acid (DHA) increases red blood cell membrane flexibility in mice with sickle cell disease.

PubMed

Wandersee, Nancy J; Maciaszek, Jamie L; Giger, Katie M; Hanson, Madelyn S; Zheng, Suilan; Guo, YiHe; Mickelson, Barbara; Hillery, Cheryl A; Lykotrafitis, George; Low, Philip S; Hogg, Neil

2015-02-01

Humans and mice with sickle cell disease (SCD) have rigid red blood cells (RBCs). Omega-3 fatty acids, such as docosahexanoic acid (DHA), may influence RBC deformability via incorporation into the RBC membrane. In this study, sickle cell (SS) mice were fed natural ingredient rodent diets supplemented with 3% DHA (DHA diet) or a control diet matched in total fat (CTRL diet). After 8weeks of feeding, we examined the RBCs for: 1) stiffness, as measured by atomic force microscopy; 2) deformability, as measured by ektacytometry; and 3) percent irreversibly sickled RBCs on peripheral blood smears. Using atomic force microscopy, it is found that stiffness is increased and deformability decreased in RBCs from SS mice fed CTRL diet compared to wild-type mice. In contrast, RBCs from SS mice fed DHA diet had markedly decreased stiffness and increased deformability compared to RBCs from SS mice fed CTRL diet. Furthermore, examination of peripheral blood smears revealed less irreversibly sickled RBCs in SS mice fed DHA diet as compared to CTRL diet. In summary, our findings indicate that DHA supplementation improves RBC flexibility and reduces irreversibly sickled cells by 40% in SS mice. These results point to potential therapeutic benefits of dietary omega-3 fatty acids in SCD. Copyright © 2014 Elsevier Inc. All rights reserved.

Diagnostic challenges and case management of the first imported case of Plasmodium knowlesi in Sri Lanka.

PubMed

Dewanee Ranaweera, A; Danansuriya, Manjula N; Pahalagedera, Kusumawathie; de A W Gunasekera, W M Kumudunayana T; Dharmawardena, Priyani; Mak, Keng Wai; Wong, Pei-Sze Jeslyn; Li, Mei-Zhi Irene; Tan, Cheong Huat; Hapuarachchi, Hapuarachchige C; Herath, Hema D B; Fernando, Deepika

2017-03-21

Sri Lanka has achieved 'malaria-free' status and is now in the phase of prevention of re-introduction of malaria. Imported malaria remains a challenge to resurgence of the disease. The diagnostic challenges encountered and the rapid response initiated to manage a Plasmodium infection, which was later confirmed as Plasmodium knowlesi, the first reported case from Sri Lanka, is discussed. An army officer who returned from Malaysia in October 2016 was found to be positive for Plasmodium both by microscopy and rapid diagnostic test (RDT) by the Anti Malaria Campaign Sri Lanka (AMC) during his third visit to a health care provider. Microscopy findings were suspicious of P. knowlesi infection as the smears showed parasite stages similar to both Plasmodium malariae and Plasmodium falciparum. Nested PCR at AMC confirmed Plasmodium genus, but not the species. In the absence of species confirmation, the patient was treated as a case of P. falciparum. The presence of P. knowlesi was later confirmed by a semi-nested PCR assay performed at the Environmental Health Institute, National Environmental Agency in Singapore. The parasite strain was also characterized by sequencing the circumsporozoite gene. Extensive case investigation including parasitological and entomological surveillance was carried out. Plasmodium knowlesi should be suspected in patients returning from countries in the South Asian region where the parasite is prevalent and when blood smear results are inconclusive.

Evaluating the diagnostic accuracy of the Xpert MTB/RIF assay on bronchoalveolar lavage fluid: A retrospective study.

PubMed

Lu, Yanjun; Zhu, Yaowu; Shen, Na; Tian, Lei; Sun, Ziyong

2018-02-08

Limited data on the diagnostic accuracy of the Xpert MTB/RIF assay using bronchoalveolar lavage fluid from patients with suspected pulmonary tuberculosis (PTB) have been reported in China. Therefore, a retrospective study was designed to evaluate the diagnostic accuracy of this assay. Clinical, radiological, and microbiological characteristics of 238 patients with suspected PTB were reviewed retrospectively. The sensitivity, specificity, positive predictive value, and negative predictive value for the diagnosis of active PTB were calculated for the Xpert MTB/RIF assay using TB culture or final diagnosis based on clinical and radiological evaluation as the reference standard. The sensitivity and specificity of the Xpert MTB/RIF assay were 84.5% and 98.9%, respectively, and those for smear microscopy were 36.2% and 100%, respectively, when compared to the culture method. However, compared with the sensitivity and specificity of final diagnosis based on clinical and radiological evaluation, the sensitivity and specificity of the assay were 72.9% and 98.7%, respectively, which were significantly higher than those for smear microscopy. The Xpert MTB/RIF assay on bronchoalveolar lavage fluid could serve as an additional rapid diagnostic tool for PTB in a high TB-burden country and improve the time to TB treatment initiation in patients with PTB. Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

The hematology of captive Bobtail lizards (Tiliqua rugosa): blood counts, light microscopy, cytochemistry, and ultrastructure.

PubMed

Moller, Cheryl A; Gaál, Tibor; Mills, Jennifer N

2016-12-01

Bobtail lizards (Tiliqua rugosa) are native to Australia. The only previous study on the hematology of this species documented just 6 animals. The aims of this study were to characterize the light microscopy, ultrastructure and cytochemistry of blood cells, and evaluate CBCs of captive Bobtail lizards. Over 2 consecutive summers, heparinized venous blood was collected from the ventral coccygeal vein of 46 clinically healthy, captive indoor- or outdoor-housed adult Bobtails. Complete blood cell counts and smear evaluations were performed, and cytochemical stains and transmission electron microscopy were used to further characterize blood cells. The eosinophils of this species were uniformly vacuolated: a unique feature not previously reported in reptiles. Heterophils were the predominant leukocyte, with fewer lymphocytes, azurophilic and nonazurophilic monocytes, occasional eosinophils, and basophils. Thrombocytes were frequently clumped. Slight polychromasia (0-15% of erythrocytes) was typically present. Hemogregarine parasites were seen on some smears. The range of CBC results was often wide. The PCV ranged from 11% to 38%. Total plasma proteins by refractometry were between 3.5 and 7.8 g/dL. Hemoglobin ranged between 2.6 and 12.6 g/dL by the modified hemoglobin-hydroxylamine method. Manual RBC count was 0.35-1.27 × 10 6 /μL, and WBC count was 2.86-22.66 × 10 3 /μL. Bobtail lizards housed outdoors had lower PCVs than indoor-housed animals. Bobtails with hemogregarine infections had lower PCVs than noninfected lizards. Ranges for CBC data were often very wide, influenced by preanalytic and analytic factors. Hemogregarine infection is associated with a decreased PCV, suggesting that some hemogregarine species are pathogenic in this population. © 2016 American Society for Veterinary Clinical Pathology.

Malaria Diagnosis Using a Mobile Phone Polarized Microscope

NASA Astrophysics Data System (ADS)

Pirnstill, Casey W.; Coté, Gerard L.

2015-08-01

Malaria remains a major global health burden, and new methods for low-cost, high-sensitivity, diagnosis are essential, particularly in remote areas with low-resource around the world. In this paper, a cost effective, optical cell-phone based transmission polarized light microscope system is presented for imaging the malaria pigment known as hemozoin. It can be difficult to determine the presence of the pigment from background and other artifacts, even for skilled microscopy technicians. The pigment is much easier to observe using polarized light microscopy. However, implementation of polarized light microscopy lacks widespread adoption because the existing commercial devices have complicated designs, require sophisticated maintenance, tend to be bulky, can be expensive, and would require re-training for existing microscopy technicians. To this end, a high fidelity and high optical resolution cell-phone based polarized light microscopy system is presented which is comparable to larger bench-top polarized microscopy systems but at much lower cost and complexity. The detection of malaria in fixed and stained blood smears is presented using both, a conventional polarized microscope and our cell-phone based system. The cell-phone based polarimetric microscopy design shows the potential to have both the resolution and specificity to detect malaria in a low-cost, easy-to-use, modular platform.

Malaria Diagnosis Using a Mobile Phone Polarized Microscope

PubMed Central

Pirnstill, Casey W.; Coté, Gerard L.

2015-01-01

Malaria remains a major global health burden, and new methods for low-cost, high-sensitivity, diagnosis are essential, particularly in remote areas with low-resource around the world. In this paper, a cost effective, optical cell-phone based transmission polarized light microscope system is presented for imaging the malaria pigment known as hemozoin. It can be difficult to determine the presence of the pigment from background and other artifacts, even for skilled microscopy technicians. The pigment is much easier to observe using polarized light microscopy. However, implementation of polarized light microscopy lacks widespread adoption because the existing commercial devices have complicated designs, require sophisticated maintenance, tend to be bulky, can be expensive, and would require re-training for existing microscopy technicians. To this end, a high fidelity and high optical resolution cell-phone based polarized light microscopy system is presented which is comparable to larger bench-top polarized microscopy systems but at much lower cost and complexity. The detection of malaria in fixed and stained blood smears is presented using both, a conventional polarized microscope and our cell-phone based system. The cell-phone based polarimetric microscopy design shows the potential to have both the resolution and specificity to detect malaria in a low-cost, easy-to-use, modular platform. PMID:26303238

Evaluation of the interaction between sodium hypochlorite and several formulations containing chlorhexidine and its effect on the radicular dentin--SEM and push-out bond strength analysis.

PubMed

Graziele Magro, Miriam; Kuga, Milton Carlos; Regina Victorino, Keli; Vázquez-Garcia, Fernando Antonio; Aranda-Garcia, Arturo Javier; Faria-Junior, Norberto Batista; Faria, Gisele; Luis Shinohara, André

2014-01-01

The aim of the current study was to evaluate the presence of debris and smear layer after endodontic irrigation with different formulations of 2% chlorhexidine gluconate (CHX) and its effects on the push-out bond strength of an epoxy-based sealer on the radicular dentin. One hundred extracted human canines were prepared to F5 instrument and irrigated with 2.5% sodium hypochlorite and 17% ethylenediaminetetraacetic acid. Fifty teeth were divided into five groups (n = 10), according to the final irrigation protocol with different 2% CHX formulations: G1 (control, no final rinse irrigation), G2 (CHX solution), G3 (CHX gel), G4 (Concepsis), and G5 (CHX Plus). In sequence, the specimens were submitted to scanning electron microscopy (SEM) analysis, in the cervical-medium and medium-apical segments, to evaluate the presence of debris and smear layer. The other 50 teeth were treated equally to a SEM study, but with the root canals filled with an epoxy-based endodontic sealer and submitted to a push-out bond strength test, in the cervical, middle, and apical thirds. G2, G3, G4, and G5 provided higher precipitation of the debris and smear layer than G1 (P < 0.05), but these groups were similar to each other (P > 0.05), in both segments. The values obtained in the push out test did not differ between groups, independent of the radicular third (P > 0.05). The CHXs formulations caused precipitation of the debris and smear layer on the radicular dentin, but these residues did not interfere in the push-out bond strength of the epoxy-based sealer. Copyright © 2013 Wiley Periodicals, Inc.

A comparison of residual smear layer and erosion following different endodontic irrigation protocols tested under clinical and laboratory conditions.

PubMed

Cehreli, Zafer C; Uyanik, M Ozgur; Nagas, Emre; Tuncel, Behram; Er, Nuray; Comert, Fugen Dagli

2013-09-01

To compare the smear layer removal efficacy and erosive effects of different irrigation protocols under clinical and laboratory conditions. Mandibular third molars (n = 32) of 30-45 year-old patients were instrumented with rotary files and were randomly assigned to one of the following groups for final irrigation: (1) 5.25% NaOCl; (2) 17% EDTA; and (3) BioPure MTAD. Thereafter, the teeth were immediately extracted and processed for micromorphological investigation. In vitro specimen pairs were prepared by repeating the clinical experiments on freshly-extracted mandibular third molars. To compare open and closed systems, laboratory experiments were repeated on 32 additional teeth with enlarged apical foramen. The cleanliness of the root canals and the extent of erosion were assessed by environmental scanning electron microscopy. Specimens prepared under clinical and laboratory conditions had similar cleanliness and erosion scores (p > 0.05). Under both conditions, the tested solutions were more effective in removing the smear layer in the coronal and middle regions than in the apical one. Comparison of closed and open systems showed similar levels of cleanliness and erosion in all regions (p > 0.05), with the exception of 17% EDTA showing significantly higher levels of cleanliness and erosion in the apical third of open-end specimens. Based on clinical correlates of in vitro root canal cleanliness and erosion, laboratory testing of root canal irrigants on extracted teeth with closed apices can serve as a reliable method to simulate the clinical condition. EDTA was the most effective final irrigation solution in removing the smear layer at the expense of yielding the greatest erosive effect.

Effect of a new desensitizing material on human dentin permeability.

PubMed

Rusin, Richard P; Agee, Kelli; Suchko, Michael; Pashley, David H

2010-06-01

Resin-modified glass ionomers (RMGI) have demonstrated clinical success providing immediate and long-term relief from root sensitivity. RMGIs have been recently introduced as paste-liquid systems for convenience of clinical usage. The objective of this study was to measure the ability of a new paste-liquid RMGI to reduce fluid flow through human dentin, compared to an established single-bottle nanofilled total etch resin adhesive indicated for root desensitization. Dentin permeability was measured on human crown sections on etched dentin, presenting a model for the exposed tubules typical of root sensitivity, and permitting measurement of the maximum permeability. In the first two groups, the etched dentin was coated with either the RMGI or adhesive, and permeability measured on the coated dentin. In a third group, a smear layer was created on the dentin with sandpaper, then the specimens were coated with the RMGI; permeability was measured on the smeared and coated dentin. Specimens from each group were sectioned and examined via scanning electron microscopy (SEM). Both the resin adhesive and the new paste-liquid RMGI protective material significantly reduced fluid flow through dentin, and exhibited excellent seal on dentin with either open tubules or smear-layer occluded tubules. The RMGI infiltrated the smear layer with resin during placement, penetrated dentin tubules, and formed resin tags. The RMGI was equivalent to the adhesive in its ability to reduce fluid flow and seal dentin. It is therefore concluded that the new RMGI and the adhesive show the potential to offer excellent sensitivity relief on exposed root dentin. Copyright (c) 2010 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Fragmented red cells reference range (Sysmex XN(®) automated blood cell counter).

PubMed

Lesesve, Jean-François; Daigney, Amandine; Henry, Sylvain; Speyer, Elodie

2015-01-01

Fragmented red cells (FRCs) is a new parameter automatedly determined by recent blood cell counters. Their count might be of interest because FRCs are supposed to reflect schistocytes counts measured on a stained peripheral blood smear observed under the microscope. But FRCs depend from the technical procedure used to detect them and thus reference ranges are device-dependent. The XN-9000(®) is one of the last model from Sysmex series. We aimed to establish reference range for FRCs, from 2389 controls. The mean ± SD was 0.32% ± 0.81, the median 0.02% (95% confidence interval ot the mean: 0.29-0.35%). We observed that the percentage of red blood cells with less than 17 pg of hemoglobin content (Hypo-He) was correlated to FRC increase, Hypo-He increase resulting in spurious FRCs majoration. FRCs reference range should be useful for: 1) laboratory staff in order to select which blood smears to check optically; 2) Sysmex company to set-up more optimal rules proposed with the counter (automated making of blood smear).

Smeared spectrum jamming suppression based on generalized S transform and threshold segmentation

NASA Astrophysics Data System (ADS)

Li, Xin; Wang, Chunyang; Tan, Ming; Fu, Xiaolong

2018-04-01

Smeared Spectrum (SMSP) jamming is an effective jamming in countering linear frequency modulation (LFM) radar. According to the time-frequency distribution difference between jamming and echo, a jamming suppression method based on Generalized S transform (GST) and threshold segmentation is proposed. The sub-pulse period is firstly estimated based on auto correlation function firstly. Secondly, the time-frequency image and the related gray scale image are achieved based on GST. Finally, the Tsallis cross entropy is utilized to compute the optimized segmentation threshold, and then the jamming suppression filter is constructed based on the threshold. The simulation results show that the proposed method is of good performance in the suppression of false targets produced by SMSP.

Impact of point-of-care implementation of Xpert® MTB/RIF: product vs. process innovation.

PubMed

Schumacher, S G; Thangakunam, B; Denkinger, C M; Oliver, A A; Shakti, K B; Qin, Z Z; Michael, J S; Luo, R; Pai, M; Christopher, D J

2015-09-01

Both product innovation (e.g., more sensitive tests) and process innovation (e.g., a point-of-care [POC] testing programme) could improve patient outcomes. To study the respective contributions of product and process innovation in improving patient outcomes. We implemented a POC programme using Xpert(®) MTB/RIF in an out-patient clinic of a tertiary care hospital in India. We measured the impact of process innovation by comparing time to diagnosis with routine testing vs. POC testing. We measured the impact of product innovation by comparing accuracy and time to diagnosis using smear microscopy vs. POC Xpert. We enrolled 1012 patients over a 15-month period. Xpert had high accuracy, but the incremental value of one Xpert over two smears was only 6% (95%CI 3-12). Implementing Xpert as a routine laboratory test did not reduce the time to diagnosis compared to smear-based diagnosis. In contrast, the POC programme reduced the time to diagnosis by 5.5 days (95%CI 4.3-6.7), but required dedicated staff and substantial adaptation of clinic workflow. Process innovation by way of a POC Xpert programme had a greater impact on time to diagnosis than the product per se, and can yield important improvements in patient care that are complementary to those achieved by introducing innovative technologies.

Performance of a Highly Sensitive Mycobacterium tuberculosis Complex Real-Time PCR Assay for Diagnosis of Pulmonary Tuberculosis in a Low-Prevalence Setting: a Prospective Intervention Study.

PubMed

Vinuesa, Víctor; Borrás, Rafael; Briones, María Luisa; Clari, María Ángeles; Cresencio, Vicenta; Giménez, Estela; Muñoz, Carmen; Oltra, Rosa; Servera, Emilio; Scheelje, Talia; Tornero, Carlos; Navarro, David

2018-05-01

The potential impact of routine real-time PCR testing of respiratory specimens from patients with presumptive tuberculosis in terms of diagnostic accuracy and time to tuberculosis treatment inception in low-prevalence settings remains largely unexplored. We conducted a prospective intervention cohort study. Respiratory specimens from 1,020 patients were examined by acid-fast bacillus smear microscopy, tested by a real-time Mycobacterium tuberculosis complex PCR assay (Abbott RealTi me MTB PCR), and cultured in mycobacterial media. Seventeen patients tested positive by PCR (5 were acid-fast bacillus smear positive and 12 acid-fast bacillus smear negative), and Mycobacterium tuberculosis was recovered from cultures for 12 of them. Patients testing positive by PCR and negative by culture ( n = 5) were treated and deemed to have responded to antituberculosis therapy. There were no PCR-negative/culture-positive cases, and none of the patients testing positive for nontuberculous mycobacteria ( n = 20) yielded a positive PCR result. The data indicated that routine testing of respiratory specimens from patients with presumptive tuberculosis by the RealTi me MTB PCR assay improves the tuberculosis diagnostic yield and may reduce the time to antituberculosis treatment initiation. On the basis of our data, we propose a novel mycobacterial laboratory algorithm for tuberculosis diagnosis. Copyright © 2018 American Society for Microbiology.

Towards a precise test for malaria diagnosis in the Brazilian Amazon: comparison among field microscopy, a rapid diagnostic test, nested PCR, and a computational expert system based on artificial neural networks

PubMed Central

2010-01-01

Background Accurate malaria diagnosis is mandatory for the treatment and management of severe cases. Moreover, individuals with asymptomatic malaria are not usually screened by health care facilities, which further complicates disease control efforts. The present study compared the performances of a malaria rapid diagnosis test (RDT), the thick blood smear method and nested PCR for the diagnosis of symptomatic malaria in the Brazilian Amazon. In addition, an innovative computational approach was tested for the diagnosis of asymptomatic malaria. Methods The study was divided in two parts. For the first part, passive case detection was performed in 311 individuals with malaria-related symptoms from a recently urbanized community in the Brazilian Amazon. A cross-sectional investigation compared the diagnostic performance of the RDT Optimal-IT, nested PCR and light microscopy. The second part of the study involved active case detection of asymptomatic malaria in 380 individuals from riverine communities in Rondônia, Brazil. The performances of microscopy, nested PCR and an expert computational system based on artificial neural networks (MalDANN) using epidemiological data were compared. Results Nested PCR was shown to be the gold standard for diagnosis of both symptomatic and asymptomatic malaria because it detected the major number of cases and presented the maximum specificity. Surprisingly, the RDT was superior to microscopy in the diagnosis of cases with low parasitaemia. Nevertheless, RDT could not discriminate the Plasmodium species in 12 cases of mixed infections (Plasmodium vivax + Plasmodium falciparum). Moreover, the microscopy presented low performance in the detection of asymptomatic cases (61.25% of correct diagnoses). The MalDANN system using epidemiological data was worse that the light microscopy (56% of correct diagnoses). However, when information regarding plasma levels of interleukin-10 and interferon-gamma were inputted, the MalDANN performance sensibly increased (80% correct diagnoses). Conclusions An RDT for malaria diagnosis may find a promising use in the Brazilian Amazon integrating a rational diagnostic approach. Despite the low performance of the MalDANN test using solely epidemiological data, an approach based on neural networks may be feasible in cases where simpler methods for discriminating individuals below and above threshold cytokine levels are available. PMID:20459613

Towards a precise test for malaria diagnosis in the Brazilian Amazon: comparison among field microscopy, a rapid diagnostic test, nested PCR, and a computational expert system based on artificial neural networks.

PubMed

Andrade, Bruno B; Reis-Filho, Antonio; Barros, Austeclino M; Souza-Neto, Sebastião M; Nogueira, Lucas L; Fukutani, Kiyoshi F; Camargo, Erney P; Camargo, Luís M A; Barral, Aldina; Duarte, Angelo; Barral-Netto, Manoel

2010-05-06

Accurate malaria diagnosis is mandatory for the treatment and management of severe cases. Moreover, individuals with asymptomatic malaria are not usually screened by health care facilities, which further complicates disease control efforts. The present study compared the performances of a malaria rapid diagnosis test (RDT), the thick blood smear method and nested PCR for the diagnosis of symptomatic malaria in the Brazilian Amazon. In addition, an innovative computational approach was tested for the diagnosis of asymptomatic malaria. The study was divided in two parts. For the first part, passive case detection was performed in 311 individuals with malaria-related symptoms from a recently urbanized community in the Brazilian Amazon. A cross-sectional investigation compared the diagnostic performance of the RDT Optimal-IT, nested PCR and light microscopy. The second part of the study involved active case detection of asymptomatic malaria in 380 individuals from riverine communities in Rondônia, Brazil. The performances of microscopy, nested PCR and an expert computational system based on artificial neural networks (MalDANN) using epidemiological data were compared. Nested PCR was shown to be the gold standard for diagnosis of both symptomatic and asymptomatic malaria because it detected the major number of cases and presented the maximum specificity. Surprisingly, the RDT was superior to microscopy in the diagnosis of cases with low parasitaemia. Nevertheless, RDT could not discriminate the Plasmodium species in 12 cases of mixed infections (Plasmodium vivax + Plasmodium falciparum). Moreover, the microscopy presented low performance in the detection of asymptomatic cases (61.25% of correct diagnoses). The MalDANN system using epidemiological data was worse that the light microscopy (56% of correct diagnoses). However, when information regarding plasma levels of interleukin-10 and interferon-gamma were inputted, the MalDANN performance sensibly increased (80% correct diagnoses). An RDT for malaria diagnosis may find a promising use in the Brazilian Amazon integrating a rational diagnostic approach. Despite the low performance of the MalDANN test using solely epidemiological data, an approach based on neural networks may be feasible in cases where simpler methods for discriminating individuals below and above threshold cytokine levels are available.

Clinicopathologic assessment of Candida colonization of oral leukoplakia.

PubMed

Sarkar, Reena; Rathod, G P

2014-01-01

Leukoplakia is the most common premalignant lesion of the oral mucosa. We studied the colonization of Candida in oral leukoplakia using direct microscopy, culture and histopathology to determine if there is a statistical correlation between Candida invasion and the clinical appearance and presence of epithelial dysplasia in leukoplakia. Samples were collected from 40 patients with oral leukoplakia and 21 controls. The swabs collected were used to inoculate Sabouraud's dextrose agar slant and for direct microscopy with Gram's stain. Culture growths were subjected to germ tube and corn meal agar tests to differentiate between Candida albicans and non-albicans groups. Biopsies were also done in all patients for histopathological confirmation; Gomori's methanamine silver stain was used to identify fungal invasion of lesional epithelium. Nineteen cases of leukoplakia showed Candida on direct smears, compared to 3 controls. Eighteen cases and one control showed growth of Candida on culture. Non-homogenous leukoplakia showed a higher positivity rate on microscopy and culture than homogenous lesions. All these correlations were statistically significant. Forty percent of leukoplakia cases were simultaneously positive for Candida on direct microscopy, culture and histopathologic evaluation. No significant difference was found between non-dysplastic and distinctly dysplastic lesions with respect to Candida detection on microscopy or culture.

Low-cost mobile phone microscopy with a reversed mobile phone camera lens.

PubMed

Switz, Neil A; D'Ambrosio, Michael V; Fletcher, Daniel A

2014-01-01

The increasing capabilities and ubiquity of mobile phones and their associated digital cameras offer the possibility of extending low-cost, portable diagnostic microscopy to underserved and low-resource areas. However, mobile phone microscopes created by adding magnifying optics to the phone's camera module have been unable to make use of the full image sensor due to the specialized design of the embedded camera lens, exacerbating the tradeoff between resolution and field of view inherent to optical systems. This tradeoff is acutely felt for diagnostic applications, where the speed and cost of image-based diagnosis is related to the area of the sample that can be viewed at sufficient resolution. Here we present a simple and low-cost approach to mobile phone microscopy that uses a reversed mobile phone camera lens added to an intact mobile phone to enable high quality imaging over a significantly larger field of view than standard microscopy. We demonstrate use of the reversed lens mobile phone microscope to identify red and white blood cells in blood smears and soil-transmitted helminth eggs in stool samples.

Low-Cost Mobile Phone Microscopy with a Reversed Mobile Phone Camera Lens

PubMed Central

Fletcher, Daniel A.

2014-01-01

The increasing capabilities and ubiquity of mobile phones and their associated digital cameras offer the possibility of extending low-cost, portable diagnostic microscopy to underserved and low-resource areas. However, mobile phone microscopes created by adding magnifying optics to the phone's camera module have been unable to make use of the full image sensor due to the specialized design of the embedded camera lens, exacerbating the tradeoff between resolution and field of view inherent to optical systems. This tradeoff is acutely felt for diagnostic applications, where the speed and cost of image-based diagnosis is related to the area of the sample that can be viewed at sufficient resolution. Here we present a simple and low-cost approach to mobile phone microscopy that uses a reversed mobile phone camera lens added to an intact mobile phone to enable high quality imaging over a significantly larger field of view than standard microscopy. We demonstrate use of the reversed lens mobile phone microscope to identify red and white blood cells in blood smears and soil-transmitted helminth eggs in stool samples. PMID:24854188

Xpert MTB/RIF assay can be used on archived gastric aspirate and induced sputum samples for sensitive diagnosis of paediatric tuberculosis.

PubMed

Singh, Sarman; Singh, Amit; Prajapati, Suneel; Kabra, Sushil K; Lodha, Rakesh; Mukherjee, Aparna; Singh, Varinder; Hesseling, Anneke C; Grewal, Harleen M S

2015-09-29

Tuberculosis (TB) in children is neglected, mainly due to lack of sensitive diagnostic tools. Recently Xpert MTB/RIF assay has revolutionized the diagnostic field, but its usefulness in pediatric TB has not been reported from India and no report is available on its use on long term archived samples. We recruited 130 pediatric patients with probable intrathoracic tuberculosis and their gastric aspirate (GA) and induced sputum (IS) samples on 2 consecutive days were collected between January 2009 and December 2012. All samples (n = 520) were subjected to smear examination, BACTEC-MGIT culture and in-house multiplex PCR. An aliquot of each sample was stored at -80 °C and tested in Xpert MTB/RIF assay in 2013. Sample wise and patient wise detection rate of smear microscopy was 4.4 % and 10 %, while for BACTEC-MGIT culture this rate was 24.4 % and 46.9 %, respectively. Of the 130 day 1 GA samples, 31.5 % and 27.7 % day 2 GA samples were culture positive. Only 17.7 % GA samples were positive on both days. Of the 130 IS samples collected on day 1 and day 2, 15.4 % and 23.1 % samples were culture positive. A combination of GA and IS yielded best results. Combining both GA and IS, the overall sensitivity of Xpert MTB/RIF on smear and culture positive samples was 95.6 %. In smear negative and culture positive samples its sensitivity was 62.5 %. The duration of sample storage impacted the Xpert MTB/RIF test performance (p = 0.0001). In smear positive samples stored for 650-849 days, its sensitivity was 85.7 % and 77.1 % for IS and GA samples which dropped to 33.3 % and 50 %, respectively, if stored for more than 1050 days. Confirmatory diagnosis of tuberculosis particularly in children is a medical challenge. No laboratory or radiological test can reach to a satisfactory level of diagnostic sensitivity. However, in this study we found that combination of multiple samples and multiple diagnostic tests can give much better yield, though not optimum. In present study, combination of 2 gastric aspirates (GA) and 2 induced sputum (IS) samples collected on two consecutive days, and tested on three diagnostic methods yielded a significantly high detection rate. Despite long term storage, the overall sensitivity of Xpert MTB/RIF on smear and -culture positive samples remained very high. But after storing these samples under subfreezing conditions the sensitivity of Xpert MTB/RIF decreased significantly. This is expected because even if the sample is smear and culture positive, the count of surviving mycobacteria goes down, after several years this count can reach to a undetectable level. This report shows that smear and culture positive samples stored at subfreezing conditions for several years can be used in the Xpert MTB/RIF assay, while maintaining appreciable diagnostic test sensitivity and specificity.

Comparison of skin smears and biopsy specimens for demonstration of Leishmania tropica bodies in cutaneous leishmaniasis.

PubMed

Dar, Nasser Rashid; Khurshid, Tariq

2005-12-01

To determine and compare the diagnostic value of skin biopsy, saline aspirate smears, skin slit smears and skin biopsy impression smears to demonstrate Leishmania tropica (LT) bodies in cutaneous leishmaniasis and to determine any association within the applied diagnostic tests. An analytical study. Department of Dermatology, PNS Shifa, Karachi from January to December 2003. Seventyeight patients of cutaneous leishmaniasis were subjected to saline aspirate smears, skin slit smears, skin biopsy impression smears and skin biopsy and looked for LT bodies. The percentage positivity of all methods was calculated. McNemar test was applied to determine the association between percentage positivities of methods. Skin biopsy showed LT bodies in 70 (89.74%) patients, saline aspirate smears in 24 (30.76%), skin slit smears in 25 (32.05%) and skin biopsy impression smears in 27 (34.61%) patients. Smears demonstrated LT bodies in 10.25% cases in which biopsy was negative. Skin biopsy impression smears showed concordant results with skin biopsy in 20.51%, skin slit smears in 16.66% and saline aspirate smears in 19.25% of cases. Saline aspirate smears were positive in 48.1% of cases in which biopsy impression smears were negative, slit smears were positive in 50.9 % of cases in which biopsy impression smears were negative and slit smears were positive in 37% of cases in which saline aspirate smears were negative. Skin biopsy for histopathology is the most useful method in the demonstration of Leishmania parasite. Sensitivity of smears can be increased by a combination of various skin smears.

Optimizing identification of clinically relevant Gram-positive organisms by use of the Bruker Biotyper matrix-assisted laser desorption ionization-time of flight mass spectrometry system.

PubMed

McElvania Tekippe, Erin; Shuey, Sunni; Winkler, David W; Butler, Meghan A; Burnham, Carey-Ann D

2013-05-01

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) can be used as a method for the rapid identification of microorganisms. This study evaluated the Bruker Biotyper (MALDI-TOF MS) system for the identification of clinically relevant Gram-positive organisms. We tested 239 aerobic Gram-positive organisms isolated from clinical specimens. We evaluated 4 direct-smear methods, including "heavy" (H) and "light" (L) smears, with and without a 1-μl direct formic acid (FA) overlay. The quality measure assigned to a MALDI-TOF MS identification is a numerical value or "score." We found that a heavy smear with a formic acid overlay (H+FA) produced optimal MALDI-TOF MS identification scores and the highest percentage of correctly identified organisms. Using a score of ≥2.0, we identified 183 of the 239 isolates (76.6%) to the genus level, and of the 181 isolates resolved to the species level, 141 isolates (77.9%) were correctly identified. To maximize the number of correct identifications while minimizing misidentifications, the data were analyzed using a score of ≥1.7 for genus- and species-level identification. Using this score, 220 of the 239 isolates (92.1%) were identified to the genus level, and of the 181 isolates resolved to the species level, 167 isolates (92.2%) could be assigned an accurate species identification. We also evaluated a subset of isolates for preanalytic factors that might influence MALDI-TOF MS identification. Frequent subcultures increased the number of unidentified isolates. Incubation temperatures and subcultures of the media did not alter the rate of identification. These data define the ideal bacterial preparation, identification score, and medium conditions for optimal identification of Gram-positive bacteria by use of MALDI-TOF MS.

Optimizing Identification of Clinically Relevant Gram-Positive Organisms by Use of the Bruker Biotyper Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry System

PubMed Central

McElvania TeKippe, Erin; Shuey, Sunni; Winkler, David W.; Butler, Meghan A.

2013-01-01

Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) can be used as a method for the rapid identification of microorganisms. This study evaluated the Bruker Biotyper (MALDI-TOF MS) system for the identification of clinically relevant Gram-positive organisms. We tested 239 aerobic Gram-positive organisms isolated from clinical specimens. We evaluated 4 direct-smear methods, including “heavy” (H) and “light” (L) smears, with and without a 1-μl direct formic acid (FA) overlay. The quality measure assigned to a MALDI-TOF MS identification is a numerical value or “score.” We found that a heavy smear with a formic acid overlay (H+FA) produced optimal MALDI-TOF MS identification scores and the highest percentage of correctly identified organisms. Using a score of ≥2.0, we identified 183 of the 239 isolates (76.6%) to the genus level, and of the 181 isolates resolved to the species level, 141 isolates (77.9%) were correctly identified. To maximize the number of correct identifications while minimizing misidentifications, the data were analyzed using a score of ≥1.7 for genus- and species-level identification. Using this score, 220 of the 239 isolates (92.1%) were identified to the genus level, and of the 181 isolates resolved to the species level, 167 isolates (92.2%) could be assigned an accurate species identification. We also evaluated a subset of isolates for preanalytic factors that might influence MALDI-TOF MS identification. Frequent subcultures increased the number of unidentified isolates. Incubation temperatures and subcultures of the media did not alter the rate of identification. These data define the ideal bacterial preparation, identification score, and medium conditions for optimal identification of Gram-positive bacteria by use of MALDI-TOF MS. PMID:23426925

Feasibility of quantitatively diagnosing cornea infection using Raman spectroscopy (Conference Presentation)

NASA Astrophysics Data System (ADS)

Bai, Yanru; Chen, Keren; Mishra, Arti; Beuerman, Roger; Liu, Quan

2017-02-01

Ocular infection is a serious eye disease that could lead to blindness without prompt and proper treatment. In pathology, ocular infection is caused by microorganisms such as bacteria, fungi or viruses. The essential prerequisite for the optimal treatment of ocular infection is to identify the microorganism causing infection early as each type of microorganism requires a different therapeutic approach. The clinical procedure for identifying the microorganism species causing ocular infection includes Gram staining (for bacteria)/microscopy (for fungi) and the culture of corneal surface scraping, or aqueous and vitreous smear samples taken from the surface of infected eyes. The culture procedure is labor intensive and expensive. Moreover, culturing is time consuming, which usually takes a few days or even weeks. Such a long delay in diagnosis could result in the exacerbation of patients' symptoms, the missing of the optimal time frame for initiating treatment and subsequently the rising cost for disease management. Raman spectroscopy has been shown highly effective for non-invasive identification of both fungi and bacteria qualitatively. In this study, we investigate the feasibility of identifying the microorganisms of ocular infection and quantifying the concentrations using Raman spectroscopy by measuring not only gram negative and gram positive bacteria but also infected cornea. By applying a modified orthogonal projection approach, the relative concentration of each bacteria species could be quantified. Our results indicate the great potential of Raman spectroscopy as an alternative tool for non-invasive diagnosis of ocular infection and could play a significantly role in future ophthalmology.

Evaluation of Polymerase Chain Reaction (PCR) with Slit Skin Smear Examination (SSS) to Confirm Clinical Diagnosis of Leprosy in Eastern Nepal

PubMed Central

Rai, Keshav; Bhattarai, Narayan Raj; Agarwal, Sudha; Khanal, Basudha

2016-01-01

Background Detection of Mycobacterium leprae in slit skin smear (SSS) is a gold standard technique for the leprosy diagnosis. Over recent years, molecular diagnosis by using PCR has been increasingly used as an alternative for its diagnosis due to its higher sensitivity. This study was carried out for comparative evaluation of PCR and SSS microscopy in a cohort of new leprosy cases diagnosed in B. P. Koirala Institute of health Sciences, Dharan, Nepal. Methodology/Principal Findings In this prospective crossectional study, 50 new clinically diagnosed cases of leprosy were included. DNA was extracted from SSS and PCR was carried out to amplify 129 bp sequence of M. leprae repetitive element. Sensitivity of SSS and PCR was 18% and 72% respectively. Improvement of 54% case detection by PCR clearly showed its advantage over SSS. Furthermore, PCR could confirm the leprosy diagnosis in 66% of AFB negative cases indicating its superiority over SSS. In the paucibacillary (PB) patients, whose BI was zero; sensitivity of PCR was 44%, whereas it was 78% in the multibacillary patients. Conclusions/Significance Our study showed PCR to be more sensitive than SSS microscopy in diagnosing leprosy. Moreover, it explored the characteristic feature of PCR which detected higher level of early stage(PB) cases tested negative by SSS. Being an expensive technique, PCR may not be feasible in all the cases, however, it would be useful in diagnosis of early cases of leprosy as opposed to SSS. PMID:28027305

Evaluation of Polymerase Chain Reaction (PCR) with Slit Skin Smear Examination (SSS) to Confirm Clinical Diagnosis of Leprosy in Eastern Nepal.

PubMed

Siwakoti, Shraddha; Rai, Keshav; Bhattarai, Narayan Raj; Agarwal, Sudha; Khanal, Basudha

2016-12-01

Detection of Mycobacterium leprae in slit skin smear (SSS) is a gold standard technique for the leprosy diagnosis. Over recent years, molecular diagnosis by using PCR has been increasingly used as an alternative for its diagnosis due to its higher sensitivity. This study was carried out for comparative evaluation of PCR and SSS microscopy in a cohort of new leprosy cases diagnosed in B. P. Koirala Institute of health Sciences, Dharan, Nepal. In this prospective crossectional study, 50 new clinically diagnosed cases of leprosy were included. DNA was extracted from SSS and PCR was carried out to amplify 129 bp sequence of M. leprae repetitive element. Sensitivity of SSS and PCR was 18% and 72% respectively. Improvement of 54% case detection by PCR clearly showed its advantage over SSS. Furthermore, PCR could confirm the leprosy diagnosis in 66% of AFB negative cases indicating its superiority over SSS. In the paucibacillary (PB) patients, whose BI was zero; sensitivity of PCR was 44%, whereas it was 78% in the multibacillary patients. Our study showed PCR to be more sensitive than SSS microscopy in diagnosing leprosy. Moreover, it explored the characteristic feature of PCR which detected higher level of early stage(PB) cases tested negative by SSS. Being an expensive technique, PCR may not be feasible in all the cases, however, it would be useful in diagnosis of early cases of leprosy as opposed to SSS.

Field evaluation of the Meade Readiview handheld microscope for diagnosis of intestinal schistosomiasis in Ugandan school children.

PubMed

Stothard, J Russell; Kabatereine, Narcis B; Tukahebwa, Edridah M; Kazibwe, Francis; Mathieson, William; Webster, Joanne P; Fenwick, Alan

2005-11-01

A novel, inexpensive handheld microscope, the Meade Readiview, was evaluated for field diagnosis of intestinal schistosomiasis by comparison of sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) against conventional compound microscopy as part of a parasitologic survey in nine sentinel schools and a rapid mapping survey across 22 schools in Uganda. Fecal smears from 685 primary school children were examined and the overall prevalence of Schistosoma mansoni was 45%. However, prevalence by school ranged widely from 0% to 100%. For individual diagnosis the Readiview had a sensitivity of 85%, a specificity of 96%, a PPV of 95%, and an NPV of 88%. Due to the poorer movement control of the glass slide on the Readiview stage, fecal smears with less than four eggs could be overlooked. At the highest magnification (160x), egg-like objects could be confounding. Estimating prevalence by school was usually within +/- 7% of that of conventional microscopy. Since the Readiview is more robust and portable, both in size and weight, and one-tenth as expensive as the traditional compound microscope, a change in the logistics and costs associated with field infection surveillance is possible. This inexpensive microscope is a pragmatic alternative to the compound microscope. It could play an important role in the collection of prevalence data to better guide anthelmintic drug delivery and also empower the diagnostic capacity of peripheral health centers where compound microscopes are few or absent.

Use of Occult Blood Detection Cards for Real-Time PCR-Based Diagnosis of Schistosoma Mansoni Infection

PubMed Central

Schunk, Mirjam; Kebede Mekonnen, Seleshi; Wondafrash, Beyene; Mengele, Carolin; Fleischmann, Erna; Herbinger, Karl-Heinz; Verweij, Jaco J.; Geldmacher, Christof; Bretzel, Gisela; Löscher, Thomas; Zeynudin, Ahmed

2015-01-01

Background In Schistosoma mansoni infection, diagnosis and control after treatment mainly rely on parasitological stool investigations which are laborious and have limited sensitivity. PCR methods have shown equal or superior sensitivity but preservation and storage methods limit their use in the field. Therefore, the use of occult blood detection cards (fecal cards) for easy sampling and storage of fecal samples for further PCR testing was evaluated in a pilot study. Methodology Stool specimens were collected in a highly endemic area for S. mansoni in Ethiopia and submitted in an investigator-blinded fashion to microscopic examination by Kato-Katz thick smear as well as to real-time PCR using either fresh frozen stool samples or stool smears on fecal cards which have been stored at ambient temperature for up to ten months. Principal Findings Out of 55 stool samples, 35 were positive by microscopy, 33 and 32 were positive by PCR of frozen samples and of fecal card samples, respectively. When microscopy was used as diagnostic “gold standard”, the sensitivity of PCR on fresh stool was 94.3% (95%-CI: 86.6; 100) and on fecal cards 91.4% (95%-CI: 82.2; 100). Conclusions The use of fecal cards proved to be a simple and useful method for stool collection and prolonged storage prior to PCR based diagnosis of S. mansoni infection. This technique may be a valuable approach for large scale surveillance and post treatment assessments PMID:26360049

Use of Occult Blood Detection Cards for Real-Time PCR-Based Diagnosis of Schistosoma Mansoni Infection.

PubMed

Schunk, Mirjam; Kebede Mekonnen, Seleshi; Wondafrash, Beyene; Mengele, Carolin; Fleischmann, Erna; Herbinger, Karl-Heinz; Verweij, Jaco J; Geldmacher, Christof; Bretzel, Gisela; Löscher, Thomas; Zeynudin, Ahmed

2015-01-01

In Schistosoma mansoni infection, diagnosis and control after treatment mainly rely on parasitological stool investigations which are laborious and have limited sensitivity. PCR methods have shown equal or superior sensitivity but preservation and storage methods limit their use in the field. Therefore, the use of occult blood detection cards (fecal cards) for easy sampling and storage of fecal samples for further PCR testing was evaluated in a pilot study. Stool specimens were collected in a highly endemic area for S. mansoni in Ethiopia and submitted in an investigator-blinded fashion to microscopic examination by Kato-Katz thick smear as well as to real-time PCR using either fresh frozen stool samples or stool smears on fecal cards which have been stored at ambient temperature for up to ten months. Out of 55 stool samples, 35 were positive by microscopy, 33 and 32 were positive by PCR of frozen samples and of fecal card samples, respectively. When microscopy was used as diagnostic "gold standard", the sensitivity of PCR on fresh stool was 94.3% (95%-CI: 86.6; 100) and on fecal cards 91.4% (95%-CI: 82.2; 100). The use of fecal cards proved to be a simple and useful method for stool collection and prolonged storage prior to PCR based diagnosis of S. mansoni infection. This technique may be a valuable approach for large scale surveillance and post treatment assessments.

Sputum smear examination and time to diagnosis in patients with smear-negative pulmonary tuberculosis in the Pacific.

PubMed

Viney, K; Bissell, K; Tabutoa, K; Kienene, T; Linh, N N; Briand, K; Harries, A D

2012-12-21

National tuberculosis programmes (NTPs) in Kiribati and the Marshall Islands, 2006-2010. To determine the proportion of all tuberculosis (TB) cases that were pulmonary smear-negative, and for these patients to determine how many sputum smears were examined and the time from sputum smear examination to registration. A retrospective cross-sectional study involving a record review of national TB and laboratory registers. Of 2420 TB cases identified, 709 (29%) were registered as smear-negative pulmonary TB. Of the 695 (98%) with information on smear examination, 222 (32%) had no smear recorded, 61 (9%) had one smear, 86 (12%) two smears and 326 (47%) three smears. Among the 473 patients who had at least one smear, 238 (50%) were registered before sputum examination, 131 (28%) within 1 week, 72 (15%) between 1 and 4 weeks, and 34 (7%) >4 weeks after sputum examination. NTPs in Kiribati and the Marshall Islands are diagnosing 29% of all TB patients as smear-negative pulmonary TB. Many patients do not have smears done or are registered before undergoing smear examination. Corrective measures are needed.

Malaria Rapid Diagnostic Tests and Malaria Microscopy for Guiding Malaria Treatment of Uncomplicated Fevers in Nigeria and Prereferral Cases in 3 African Countries.

PubMed

Falade, Catherine O; Ajayi, IkeOluwapo O; Nsungwa-Sabiiti, Jesca; Siribié, Mohamadou; Diarra, Amidou; Sermé, Luc; Afonne, Chinenye; Yusuf, Oyindamola B; Gansane, Zakaria; Jegede, Ayodele S; Singlovic, Jan; Gomes, Melba

2016-12-15

 The World Health Organization recommends that malaria treatment be based on demonstration of the infecting Plasmodium parasite specie. Malaria rapid diagnostic tests (RDTs) are recommended at community points of care because they are accurate and rapid. We report on parasitological results in a malaria study in selected rural communities in 3 African countries.  In Nigeria, community health workers (CHWs) performed RDTs (SD-Bioline) and thick blood smears on all children suspected to have malaria. Malaria RDT-positive children able to swallow received artemisinin-based combination therapy (Coartem). In all countries, children unable to take oral drugs received prereferral rectal artesunate irrespective of RDT result and were referred to the nearest health facility. Thick blood smears and RDTs were usually taken at hospital admission. In Nigeria and Burkina Faso, RDT cassettes and blood smears were re-read by an experienced investigator at study end.  Trained CHWs enrolled 2148 children in Nigeria. Complete parasitological data of 1860 (86.6%) enrollees were analyzed. The mean age of enrollees was 30.4 ± 15.7 months. The prevalence of malaria parasitemia in the study population was 77.8% (1447/1860), 77.6% (1439/1855), and 54.1% (862/1593) by RDT performed by CHWs vs an expert clinical research assistant vs microscopy (gold standard), respectively. Geometric mean parasite density was 6946/µL (range, 40-436 450/µL). There were 49 cases of RDT false-negative results with a parasite density range of 40-54 059/µL. False-negative RDT results with high parasitemia could be due to non-falciparum infection or result from a prozone effect. Sensitivity and specificity of SD-Bioline RDT results as read by CHWs were 94.3% and 41.6%, respectively, while the negative and positive predictive values were 86.1% and 65.6%, respectively. The level of agreement in RDT reading by the CHWs and experienced research staff was 86.04% and κ statistic of 0.60. The malaria parasite positivity rate by RDT and microscopy among children with danger signs in the 3 countries was 67.9% and 41.8%, respectively.  RDTs are useful in guiding malaria management and were successfully used for diagnosis by trained CHWs. However, false-negative RDT results were identified and can undermine confidence in results and control efforts. © 2016 World Health Organization; licensee Oxford Journals.

Malaria Rapid Diagnostic Tests and Malaria Microscopy for Guiding Malaria Treatment of Uncomplicated Fevers in Nigeria and Prereferral Cases in 3 African Countries

PubMed Central

Falade, Catherine O.; Ajayi, IkeOluwapo O.; Nsungwa-Sabiiti, Jesca; Siribié, Mohamadou; Diarra, Amidou; Sermé, Luc; Afonne, Chinenye; Yusuf, Oyindamola B.; Gansane, Zakaria; Jegede, Ayodele S.; Singlovic, Jan; Gomes, Melba

2016-01-01

Background. The World Health Organization recommends that malaria treatment be based on demonstration of the infecting Plasmodium parasite specie. Malaria rapid diagnostic tests (RDTs) are recommended at community points of care because they are accurate and rapid. We report on parasitological results in a malaria study in selected rural communities in 3 African countries. Methods. In Nigeria, community health workers (CHWs) performed RDTs (SD-Bioline) and thick blood smears on all children suspected to have malaria. Malaria RDT-positive children able to swallow received artemisinin-based combination therapy (Coartem). In all countries, children unable to take oral drugs received prereferral rectal artesunate irrespective of RDT result and were referred to the nearest health facility. Thick blood smears and RDTs were usually taken at hospital admission. In Nigeria and Burkina Faso, RDT cassettes and blood smears were re-read by an experienced investigator at study end. Results. Trained CHWs enrolled 2148 children in Nigeria. Complete parasitological data of 1860 (86.6%) enrollees were analyzed. The mean age of enrollees was 30.4 ± 15.7 months. The prevalence of malaria parasitemia in the study population was 77.8% (1447/1860), 77.6% (1439/1855), and 54.1% (862/1593) by RDT performed by CHWs vs an expert clinical research assistant vs microscopy (gold standard), respectively. Geometric mean parasite density was 6946/µL (range, 40–436 450/µL). There were 49 cases of RDT false-negative results with a parasite density range of 40–54 059/µL. False-negative RDT results with high parasitemia could be due to non-falciparum infection or result from a prozone effect. Sensitivity and specificity of SD-Bioline RDT results as read by CHWs were 94.3% and 41.6%, respectively, while the negative and positive predictive values were 86.1% and 65.6%, respectively. The level of agreement in RDT reading by the CHWs and experienced research staff was 86.04% and κ statistic of 0.60. The malaria parasite positivity rate by RDT and microscopy among children with danger signs in the 3 countries was 67.9% and 41.8%, respectively. Conclusions. RDTs are useful in guiding malaria management and were successfully used for diagnosis by trained CHWs. However, false-negative RDT results were identified and can undermine confidence in results and control efforts. PMID:27941107

Use of a three-band HRP2/pLDH combination rapid diagnostic test increases diagnostic specificity for falciparum malaria in Ugandan children.

PubMed

Hawkes, Michael; Conroy, Andrea L; Opoka, Robert O; Namasopo, Sophie; Liles, W Conrad; John, Chandy C; Kain, Kevin C

2014-02-01

Rapid diagnostic tests (RDTs) for malaria provide a practical alternative to light microscopy for malaria diagnosis in resource-limited settings. Three-band RDTs incorporating two parasite antigens may have enhanced diagnostic specificity, relative to two-band RDTs with a single parasite antigen (typically histidine-rich protein 2 [HRP2]). Phase 1: 2,000 children, two months to five years of age, admitted to a referral hospital in Jinja, Uganda, with acute febrile illness were enrolled. A WHO highly rated three-band RDT was compared to light microscopy of thick peripheral blood films read by local expert microscopists.Phase 2: the three-band RDT was used as a screening tool for inclusion of patients in a clinical trial, and subjects with three positive RDT bands were tested by microscopy using blood samples drawn in parallel. Discordant results were adjudicated by PCR. Phase 1: 1,648 children had both a RDT and peripheral blood smear performed. The specificity of a RDT with all three bands positive was 82% (95% CI: 79-85%) compared to 62% (95% CI: 59-66%) for HRP2 alone. The sensitivity was 88% (95% CI: 85-89%) and 94% (95% CI: 92-95%) for three-band positive RDT and HRP2 antigen, respectively. 119 patients (7.2%) had a positive HRP2 band, but negative parasite lactate dehydrogenase (pLHD) band and negative peripheral smear, and 72 (61%) of these had received pre-treatment with anti-malarials, suggesting a false positive HRP2 result (p = 0.002).Phase 2: the positive predictive value (PPV) of the three-band RDT was 94% (95% CI 89%-97%) using microscopy as the reference standard. However, microscopy-discordant results were shown to be positive for P. falciparum by PCR in all cases, suggesting that the PPV was in fact higher. The pLDH antigen on three-band RDTs, used in combination with HRP2, provides added diagnostic specificity for malaria parasitaemia and may be useful to distinguish acute infection from recently treated infection. In situations where diagnostic specificity is desirable (e.g., for selection of malaria-infected participants in clinical trials), a three-band RDT should be considered in a sub-Saharan African setting.

Sputum smear examination and time to diagnosis in patients with smear-negative pulmonary tuberculosis in the Pacific

PubMed Central

Bissell, K.; Tabutoa, K.; Kienene, T.; Linh, N. N.; Briand, K.; Harries, A. D.

2012-01-01

Setting: National tuberculosis programmes (NTPs) in Kiribati and the Marshall Islands, 2006–2010. Objective: To determine the proportion of all tuberculosis (TB) cases that were pulmonary smear-negative, and for these patients to determine how many sputum smears were examined and the time from sputum smear examination to registration. Design: A retrospective cross-sectional study involving a record review of national TB and laboratory registers. Results: Of 2420 TB cases identified, 709 (29%) were registered as smear-negative pulmonary TB. Of the 695 (98%) with information on smear examination, 222 (32%) had no smear recorded, 61 (9%) had one smear, 86 (12%) two smears and 326 (47%) three smears. Among the 473 patients who had at least one smear, 238 (50%) were registered before sputum examination, 131 (28%) within 1 week, 72 (15%) between 1 and 4 weeks, and 34 (7%) >4 weeks after sputum examination. Conclusion: NTPs in Kiribati and the Marshall Islands are diagnosing 29% of all TB patients as smear-negative pulmonary TB. Many patients do not have smears done or are registered before undergoing smear examination. Corrective measures are needed. PMID:26392971

Improved diagnostic testing and malaria treatment practices in Zambia.

PubMed

Hamer, Davidson H; Ndhlovu, Micky; Zurovac, Dejan; Fox, Matthew; Yeboah-Antwi, Kojo; Chanda, Pascalina; Sipilinyambe, Naawa; Simon, Jonathon L; Snow, Robert W

2007-05-23

Improving the accuracy of malaria diagnosis with rapid antigen-detection diagnostic tests (RDTs) has been proposed as an approach for reducing overtreatment of malaria in the current era of widespread implementation of artemisinin-based combination therapy in sub-Saharan Africa. To assess the association between use of microscopy and RDT and the prescription of antimalarials. Cross-sectional, cluster sample survey, carried out between March and May 2006, of all outpatients treated during 1 working day at government and mission health facilities in 4 sentinel districts in Zambia. Proportions of patients undergoing malaria diagnostic procedures and receiving antimalarial treatment. Seventeen percent of the 104 health facilities surveyed had functional microscopy, 63% had RDTs available, and 73% had 1 or more diagnostics available. Of patients with fever (suspected malaria), 27.8% (95% confidence interval [CI], 13.1%-42.5%) treated in health facilities with malaria diagnostics were tested and 44.6% had positive test results. Of patients with negative blood smear results, 58.4% (95% CI, 36.7%-80.2%) were prescribed an antimalaria drug, as were 35.5% (95% CI, 16.0%-55.0%) of those with a negative RDT result. Of patients with fever who did not have diagnostic tests done, 65.9% were also prescribed antimalarials. In facilities with artemether-lumefantrine in stock, this antimalarial was prescribed to a large proportion of febrile patients with a positive diagnostic test result (blood smear, 75.0% [95% CI, 51.7%-98.3%]; RDT, 70.4% [95% CI, 39.3%-100.0%]), but also to some of those with a negative diagnostic test result (blood smear, 30.4% [95% CI, 8.0%-52. 9%]; RDT, 26.7% [95% CI, 5.7%-47.7%]). Despite efforts to expand the provision of malaria diagnostics in Zambia, they continue to be underused and patients with negative test results frequently receive antimalarials. Provision of new tools to reduce inappropriate use of new expensive antimalarial treatments must be accompanied by a major change in clinical treatment of patients presenting with fever but lacking evidence of malaria infection.

A Diagnostic Accuracy Study of Xpert®MTB/RIF in HIV-Positive Patients with High Clinical Suspicion of Pulmonary Tuberculosis in Lima, Peru

PubMed Central

Carriquiry, Gabriela; Otero, Larissa; González-Lagos, Elsa; Zamudio, Carlos; Sánchez, Eduardo; Nabeta, Pamela; Campos, Miguel; Echevarría, Juan; Seas, Carlos; Gotuzzo, Eduardo

2012-01-01

Background Diagnosis of pulmonary tuberculosis (TB) among human immunodeficiency virus (HIV) patients remains complex and demands easy to perform and accurate tests. Xpert®MTB/RIF (MTB/RIF) is a molecular TB diagnostic test which is rapid and convenient; the test requires minimal human resources and reports results within two hours. The majority of performance studies of MTB/RIF have been performed in high HIV burden settings, thus TB diagnostic studies among HIV patients in low HIV prevalence settings such as Peru are still needed. Methodology/Principal Findings From April 2010 to May 2011, HIV-positive patients with high clinical suspicion of TB were enrolled from two tertiary hospitals in Lima, Peru. Detection of TB by MTB/RIF was compared to a composite reference standard Löwenstein-Jensen (LJ) and liquid culture. Detection of rifampicin resistance was compared to the LJ proportion method. We included 131 patients, the median CD4 cell count was 154.5 cells/mm3 and 45 (34.4%) had TB. For TB detection among HIV patients, sensitivity of MTB/RIF was 97.8% (95% CI 88.4–99.6) (44/45); specificity was 97.7% (95% CI 91.9–99.4) (84/86); the positive predictive value was 95.7% (95% CI 85.5–98.8) (44/46); and the negative predictive value, 98.8% (95% CI 93.6–99.8) (84/85). MTB/RIF detected 13/14 smear-negative TB cases, outperforming smear microscopy [97.8% (44/45) vs. 68.9% (31/45); p = 0.0002]. For rifampicin resistance detection, sensitivity of MTB/RIF was 100% (95% CI 61.0–100.0) (6/6); specificity was 91.0% (95% CI 76.4–96.9) (30/33); the positive predictive value was 66.7% (95% CI 35.4–87.9) (6/9); and the negative predictive value was 100% (95% CI 88.7 –100.0) (30/30). Conclusions/Significance In HIV patients in our population with a high clinical suspicion of TB, MTB/RIF performed well for TB diagnosis and outperformed smear microscopy. PMID:22970271

Bio-sensing with butterfly wings: naturally occurring nano-structures for SERS-based malaria parasite detection.

PubMed

Garrett, Natalie L; Sekine, Ryo; Dixon, Matthew W A; Tilley, Leann; Bambery, Keith R; Wood, Bayden R

2015-09-07

Surface enhanced Raman scattering (SERS) is a powerful tool with great potential to provide improved bio-sensing capabilities. The current 'gold-standard' method for diagnosis of malaria involves visual inspection of blood smears using light microscopy, which is time consuming and can prevent early diagnosis of the disease. We present a novel surface-enhanced Raman spectroscopy substrate based on gold-coated butterfly wings, which enabled detection of malarial hemozoin pigment within lysed blood samples containing 0.005% and 0.0005% infected red blood cells.

A PARASITOLOGIC AND MOLECULAR SURVEY OF HEPATOZOON CANIS INFECTION IN STRAY DOGS IN NORTHEAST OF IRAN.

PubMed

Barati, Ali; Razmi, Gholamreza

2018-05-15

Canine hepatozoonosis, caused by H. canis, is a tick-borne disease in domestic and wild dogs that is transmitted by ingestion of Rhipicephalus sanguineus ticks. The aim of the study was to detect H. canis in stray dogs in Iran using blood smear examination and molecular techniques. From October 2014 to September 2015, 150 EDTA blood samples were collected from stray dogs in the northeast region of Iran. Blood smears were microscopically examined for the presence of Hepatozoon gamonts; whole blood was evaluated by PCR, with subsequent sequencing and phylogenetic analysis. Hepatozoon spp. Gamonts were observed in the neutrophils of 5/150 (3.3%) blood smears, whereas Hepatozoon spp. 18S rDNA was detected in 12/150 (8.0%) blood samples from stray dogs. There was a good agreement between microscopy and PCR methods. (Kappa= 0.756). The highest rate of infection was seasonally detected in the summer (p<0.05). The difference of frequency of Hepatozoon spp infection was not significant by gender and age factors (p>0.05). The alignment analysis of the sequenced samples showed ≥99% similarity with other nucleotide sequences of Hepatozoon spp. in GenBank. The phylogenetic tree also revealed that the nucleotide sequences in this study were clustered in the H. canis clade and different from the H. felis and H. americanum clades. According to the results, it is concluded that H. canis infection is present among dogs in northeastern region of Iran.

Prevalence of gestational, placental and congenital malaria in north-west Colombia

PubMed Central

2013-01-01

Background The frequency of pregnancy-associated malaria is increasingly being documented in American countries. In Colombia, with higher frequency of Plasmodium vivax over Plasmodium falciparum infection, recent reports confirmed gestational malaria as a serious public health problem. Thick smear examination is the gold standard to diagnose malaria in endemic settings, but in recent years, molecular diagnostic methods have contributed to elucidate the dimension of the problem of gestational malaria. The study was aimed at exploring the prevalence of gestational, placental and congenital malaria in women who delivered at the local hospitals of north-west Colombia, between June 2008 and April 2011. Methods A group of 129 parturient women was selected to explore the prevalence of gestational, placental and congenital malaria in a descriptive, prospective and transversal (prevalence) design. Diagnosis was based on the simultaneous application of two independent diagnostic tests: microscopy of thick blood smears and a polymerase chain reaction assay (PCR). Results The prevalence of gestational malaria (thick smear /PCR) was 9.1%/14.0%; placental malaria was 3.3%/16.5% and congenital malaria was absent. A history of gestational malaria during the current pregnancy was significantly associated with gestational malaria at delivery. Plasmodium vivax caused 65% of cases of gestational malaria, whereas P. falciparum caused most cases of placental malaria. Conclusions Gestational and placental malaria are a serious problem in the region, but the risk of congenital malaria is low. A history of malaria during pregnancy may be a practical indicator of infection at delivery. PMID:24053184

International standards for tuberculosis care: relevance and implications for laboratory professionals.

PubMed

Pai, M; Daley, P; Hopewell, P C

2007-04-01

On World Tuberculosis (TB) Day 2006, the International Standards for Tuberculosis Care (ISTC) was officially released and widely endorsed by several agencies and organizations. The ISTC release was the culmination of a year long global effort to develop and set internationally acceptable, evidence-based standards for tuberculosis care. The ISTC describes a widely endorsed level of care that all practitioners, public and private, should seek to achieve in managing individuals who have or are suspected of having, TB and is intended to facilitate the effective engagement of all healthcare providers in delivering high quality care for patients of all ages, including those with smear-positive, smear-negative and extra-pulmonary TB, TB caused by drug-resistant Mycobacterium tuberculosis and TB/HIV coinfection. In this article, we present the ISTC, with a special focus on the diagnostic standards and describe their implications and relevance for laboratory professionals in India and worldwide. Laboratory professionals play a critical role in ensuring that all the standards are actually met by providing high quality laboratory services for smear microscopy, culture and drug susceptibility testing and other services such as testing for HIV infection. In fact, if the ISTC is widely followed, it can be expected that there will be a greater need and demand for quality assured laboratory services and this will have obvious implications for all laboratories in terms of work load, requirement for resources and trained personnel and organization of quality assurance systems.

Evaluation of a miniature microscope objective designed for fluorescence array microscopy detection of Mycobacterium tuberculosis.

PubMed

McCall, Brian; Olsen, Randall J; Nelles, Nicole J; Williams, Dawn L; Jackson, Kevin; Richards-Kortum, Rebecca; Graviss, Edward A; Tkaczyk, Tomasz S

2014-03-01

A prototype miniature objective that was designed for a point-of-care diagnostic array microscope for detection of Mycobacterium tuberculosis and previously fabricated and presented in a proof of concept is evaluated for its effectiveness in detecting acid-fast bacteria. To evaluate the ability of the microscope to resolve submicron features and details in the image of acid-fast microorganisms stained with a fluorescent dye, and to evaluate the accuracy of clinical diagnoses made with digital images acquired with the objective. The lens prescription data for the microscope design are presented. A test platform is built by combining parts of a standard microscope, a prototype objective, and a digital single-lens reflex camera. Counts of acid-fast bacteria made with the prototype objective are compared to counts obtained with a standard microscope over matched fields of view. Two sets of 20 smears, positive and negative, are diagnosed by 2 pathologists as sputum smear positive or sputum smear negative, using both a standard clinical microscope and the prototype objective under evaluation. The results are compared to a reference diagnosis of the same sample. More bacteria are counted in matched fields of view in digital images taken with the prototype objective than with the standard clinical microscope. All diagnostic results are found to be highly concordant. An array microscope built with this miniature lens design will be able to detect M tuberculosis with high sensitivity and specificity.

Tuberculosis control in Sudan against seemingly insurmountable odds.

PubMed

El Sony, A I; Baraka, O; Enarson, D A; Bjune, G

2000-07-01

Sudan, Africa's largest and one of its poorest countries, in which civil disturbance, resource limitation and communications difficulties are substantial impediments to delivery of health services. To 1) illustrate the burden of tuberculosis; 2) review measures taken to control the disease; 3) outline the introduction of the DOTS strategy; and 4) demonstrate the trend in the output of the DOTS strategy. Published information on general health, tuberculosis and health structure provide the setting. Routine reports illustrate the trend in case notification in Sudan, and outcome of treatment by period of enrollment on treatment (cohort). Since 1992, sputum smear microscopy centres have been established in existing health facilities (179 of a total 290 targeted centres). By the end of the second quarter of 1998, 82,860 cases of tuberculosis had been reported, of whom 52% were sputum smear-positive cases. Of these, 89% had no history of previous treatment for as much as one month. The treatment outcomes for 11,000 new smear-positive cases were reported by the end of the second quarter of 1997; the proportion of notified cases for whom treatment results were available increased from 16% in 1994 to 63% in 1996. Of these, 72% were successfully treated, increasing from 62% in 1994 to 73% in 1996. Despite seemingly overwhelming odds, the DOTS strategy has been successfully commenced and is in the process of expansion throughout the country, with monitoring of the quality of diagnostic examinations and improvements in treatment outcome. Further improvement is necessary, but appears feasible.

Occurrence of filaria in domestic dogs of Samburu pastoralists in Northern Kenya and its associations with canine distemper.

PubMed

Albrechtová, Kateřina; Sedlák, Kamil; Petrželková, Klára J; Hlaváč, Jan; Mihalca, Andrei D; Lesingirian, Alison; Kanyari, Paul W N; Modrý, David

2011-12-15

Samples of blood (serum, smears and blood preserved with ethanol) were collected from dogs during a vaccination campaign in northern Kenya in the years 2006 and 2007. Blood was screened for filarial parasites using molecular and microscopy methods and sera were tested for antibodies against canine distemper virus (CDV). Parasitological examination revealed the presence of two species of canine filariae: Acanthocheilonema dracunculoides and A. reconditum. The DNA from the former species was detected in 58% dogs sampled in 2006 and 36% dogs sampled in 2007, whereas the latter was found only in 4.2% samples collected in 2007. Microfilariae were found in 33.8% blood smears collected in 2006 and 10.6% blood smears collected in 2007. The seroprevalence of CDV was 33.4% in 2006 and 11.2% in 2007. The effect of sex, age and CDV-seropositivity/seronegativity on the occurrence of A. dracunculoides was evaluated. Infection by A. dracunculoides was more common in males and in dogs with a positive antibody titer for canine distemper, but evenly distributed among different age groups. The difference in the prevalence of A. dracunculoides in two isolated mountain ranges was not statistically significant. Methodologies available for detection and determination of canine filariae are compared, underlining methodical pitfalls arising through the determination of less common filarial species. The role of single epidemiological factors and possible association between canine distemper and filariasis are discussed. Copyright © 2011 Elsevier B.V. All rights reserved.

Epithelioid variant of malignant peripheral nerve sheath tumor (malignant schwannoma) of the urinary bladder.

PubMed

Eltoum, I A; Moore, R J; Cook, W; Crowe, D R; Rodgers, W H; Siegal, G P

1999-10-01

Sarcoma represents less than 2% of all neoplasms diagnosed or recognized in effusions. Epithelioid peripheral nerve sheath tumor is a rare tumor that is difficult to differentiate from other epithelioid tumors without the use of ancillary studies. A 39-year-old paraplegic man presented with hematuria and a bladder mass that extended to involve the pelvic peritoneum. Light microscopy using hematoxylin-eosin, Papanicolaou, and immunohistochemical stains as well as transmission electron microscopy showed features of epithelioid malignant peripheral nerve sheath tumor with rhabdoid features and an accompanying eosinophilic infiltrate. Cytologic smears confirmed the similarities between the primary tumor in the bladder and the cells in the pelvic fluid and excluded the possibility of reactive changes related to postsurgical radiation. Ancillary studies were critical in narrowing the differential diagnoses and reaching the final conclusion.

A portable microscopy system for fluorescence, polarized, and brightfield imaging

NASA Astrophysics Data System (ADS)

Gordon, Paul; Wattinger, Rolla; Lewis, Cody; Venancio, Vinicius Paula; Mertens-Talcott, Susanne U.; Coté, Gerard

2018-02-01

The use of mobile phones to conduct diagnostic microscopy at the point-of-care presents intriguing possibilities for the advancement of high-quality medical care in remote settings. However, it is challenging to create a single device that can adapt to the ever-varying camera technologies in phones or that can image with the customization that multiple modalities require for applications such as malaria diagnosis. A portable multi-modal microscope system is presented that utilizes a Raspberry Pi to collect and transmit data wirelessly to a myriad of electronic devices for image analysis. The microscopy system is capable of providing to the user correlated brightfield, polarized, and fluorescent images of samples fixed on traditional microscopy slides. The multimodal diagnostic capabilities of the microscope were assessed by measuring parasitemia of Plasmodium falciparum-infected thin blood smears. The device is capable of detecting fluorescently-labeled DNA using FITC excitation (490 nm) and emission (525 nm), the birefringent P. falciparum byproduct hemozoin, and detecting brightfield absorption with a resolution of 0.78 micrometers (element 9-3 of a 1951 Air Force Target). This microscopy system is a novel portable imaging tool that may be a viable candidate for field implementation if challenges of system durability, cost considerations, and full automation can be overcome.

Comparison of LED and Conventional Fluorescence Microscopy for Detection of Acid Fast Bacilli in a Low-Incidence Setting

PubMed Central

Minion, Jessica; Pai, Madhukar; Ramsay, Andrew; Menzies, Dick; Greenaway, Christina

2011-01-01

Introduction Light emitting diode fluorescence microscopes have many practical advantages over conventional mercury vapour fluorescence microscopes, which would make them the preferred choice for laboratories in both low- and high-resource settings, provided performance is equivalent. Methods In a nested case-control study, we compared diagnostic accuracy and time required to read slides with the Zeiss PrimoStar iLED, LW Scientific Lumin, and a conventional fluorescence microscope (Leica DMLS). Mycobacterial culture was used as the reference standard, and subgroup analysis by specimen source and organism isolated were performed. Results There was no difference in sensitivity or specificity between the three microscopes, and agreement was high for all comparisons and subgroups. The Lumin and the conventional fluorescence microscope were equivalent with respect to time required to read smears, but the Zeiss iLED was significantly time saving compared to both. Conclusions Light emitting diode microscopy should be considered by all tuberculosis diagnostic laboratories, including those in high income countries, as a replacement for conventional fluorescence microscopes. Our findings provide support to the recent World Health Organization policy recommending that conventional fluorescence microscopy be replaced by light emitting diode microscopy using auramine staining in all settings where fluorescence microscopy is currently used. PMID:21811622

Reliability of rapid diagnostic test for diagnosing peripheral and placental malaria in an area of unstable malaria transmission in Eastern Sudan

PubMed Central

2013-01-01

Background Diagnosing Plasmodium falciparum malaria during pregnancy is a great challenge for clinicians because of the low density of parasites in the peripheral blood and parasite sequestration in the placenta. Nevertheless, few data on the use of malaria rapid diagnostic test (RDT) during pregnancy have been published. Methods P. falciparum infections were assessed in 156 febrile pregnant women by microscopic examination of their blood smears and by RDT and polymerase chain reactions (PCR). In addition, 150 women were assessed at the time of delivery by microscopy, RDT, PCR and placental histology investigations. The study was conducted at the Gadarif Hospital, Eastern Sudan. The SD Bioline P. f / P. v (Bio Standard Diagnostics, Gurgaon, Korea) RDT kit was evaluated in this study. Results Among the febrile pregnant women, 17 (11.0%), 26 (16.7%) and 18 (11.5%) positive cases of P. falciparum were detected by microscopy, RDT, and PCR, respectively. The sensitivity and specificity of the microscopy was 94.4% and 100%, respectively. The corresponding values for RDT evaluation were 83.3% and 92.0%, as compared with PCR as the gold standard. While there were no detected cases of malaria by microscopic examination of blood smears, 27 (18.0%), 21(14.0%) and 46 (30.7%) out of the 150 placentae investigated had P. falciparum as determined by RDT, PCR, and histology, respectively. The sensitivity and specificity for RDT was 17.4% and 81.7%, respectively. The corresponding values for PCR were 6.5% and 82.7%, where histology was used as the gold standard. Conclusions The RDT kit used in this study has poor performance for peripheral and placental P. falciparum malaria detection in this setting. Virtual slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1092363465928479 PMID:23587371

Deformation mechanisms and evolution of the microstructure of gouge in the Main Fault in Opalinus Clay in the Mont Terri rock laboratory (CH)

NASA Astrophysics Data System (ADS)

Laurich, Ben; Urai, Janos L.; Vollmer, Christian; Nussbaum, Christophe

2018-01-01

We studied gouge from an upper-crustal, low-offset reverse fault in slightly overconsolidated claystone in the Mont Terri rock laboratory (Switzerland). The laboratory is designed to evaluate the suitability of the Opalinus Clay formation (OPA) to host a repository for radioactive waste. The gouge occurs in thin bands and lenses in the fault zone; it is darker in color and less fissile than the surrounding rock. It shows a matrix-based, P-foliated microfabric bordered and truncated by micrometer-thin shear zones consisting of aligned clay grains, as shown with broad-ion-beam scanning electron microscopy (BIB-SEM) and optical microscopy. Selected area electron diffraction based on transmission electron microscopy (TEM) shows evidence for randomly oriented nanometer-sized clay particles in the gouge matrix, surrounding larger elongated phyllosilicates with a strict P foliation. For the first time for the OPA, we report the occurrence of amorphous SiO2 grains within the gouge. Gouge has lower SEM-visible porosity and almost no calcite grains compared to the undeformed OPA. We present two hypotheses to explain the origin of gouge in the Main Fault: (i) authigenic generation consisting of fluid-mediated removal of calcite from the deforming OPA during shearing and (ii) clay smear consisting of mechanical smearing of calcite-poor (yet to be identified) source layers into the fault zone. Based on our data we prefer the first or a combination of both, but more work is needed to resolve this. Microstructures indicate a range of deformation mechanisms including solution-precipitation processes and a gouge that is weaker than the OPA because of the lower fraction of hard grains. For gouge, we infer a more rate-dependent frictional rheology than suggested from laboratory experiments on the undeformed OPA.

Highly sensitive detection of human papillomavirus type 16 DNA using time-resolved fluorescence microscopy and long lifetime probes

NASA Astrophysics Data System (ADS)

Wang, Xue F.; Periasamy, Ammasi; Wodnicki, Pawel; Siadat-Pajouh, M.; Herman, Brian

1995-04-01

We have been interested in the role of Human Papillomavirus (HPV) in cervical cancer and its diagnosis; to that end we have been developing microscopic imaging and fluorescent in situ hybridization (FISH) techniques to genotype and quantitate the amount of HPV present at a single cell level in cervical PAP smears. However, we have found that low levels of HPV DNA are difficult to detect accurately because theoretically obtainable sensitivity is never achieved due to nonspecific autofluorescence, fixative induced fluorescence of cells and tissues, and autofluorescence of the optical components in the microscopic system. In addition, the absorption stains used for PAP smears are intensely autofluorescent. Autofluorescence is a rapidly decaying process with lifetimes in the range of 1-100 nsec, whereas phosphorescence and delayed fluorescence have lifetimes in the range of 1 microsecond(s) ec-10 msec. The ability to discriminate between specific fluorescence and autofluorescence in the time-domain has improved the sensitivity of diagnostic test such that they perform comparably to, or even more sensitive than radioisotopic assays. We have developed a novel time-resolved fluorescence microscope to improve the sensitivity of detection of specific molecules of interest in slide based specimens. This time-resolved fluorescence microscope is based on our recently developed fluorescence lifetime imaging microscopy (FILM) in conjunction with the use of long lifetime fluorescent labels. By using fluorescence in situ hybridization and the long lifetime probe (europium), we have demonstrated the utility of this technique for detection of HPV DNA in cervicovaginal cells. Our results indicate that the use of time-resolved fluorescence microscopy and long lifetime probes increases the sensitivity of detection by removing autofluorescence and will thus lead to improved early diagnosis of cervical cancer. Since the highly sensitive detection of DNA in clinical samples using fluorescence in situ hybridization image is useful for the diagnosis of many other type of diseases, the system we have developed should find numerous applications for the diagnosis of disease states.

Diagnosing Polyparasitism in a High-Prevalence Setting in Beira, Mozambique: Detection of Intestinal Parasites in Fecal Samples by Microscopy and Real-Time PCR.

PubMed

Meurs, Lynn; Polderman, Anton M; Vinkeles Melchers, Natalie V S; Brienen, Eric A T; Verweij, Jaco J; Groosjohan, Bernhard; Mendes, Felisberto; Mechendura, Manito; Hepp, Dagmar H; Langenberg, Marijke C C; Edelenbosch, Rosanne; Polman, Katja; van Lieshout, Lisette

2017-01-01

Many different intestinal parasite species can co-occur in the same population. However, classic diagnostic tools can only frame a particular group of intestinal parasite species. Hence, one or two tests do not suffice to provide a complete picture of infecting parasite species in a given population. The present study investigated intestinal parasitic infections in Beira, Mozambique, i.e. in the informal settlement of Inhamudima. Diagnostic accuracy of five classical microscopy techniques and real-time PCR for the detection of a broad spectrum of parasites was compared. A cross-sectional population-based survey was performed. One stool sample per participant (n = 303) was examined by direct smear, formal-ether concentration (FEC), Kato smear, Baermann method, coproculture and real-time PCR. We found that virtually all people (96%) harbored at least one helminth, and that almost half (49%) harbored three helminths or more. Remarkably, Strongyloides stercoralis infections were widespread with a prevalence of 48%, and Ancylostoma spp. prevalence was higher than that of Necator americanus (25% versus 15%), the hookworm species that is often assumed to prevail in East-Africa. Among the microscopic techniques, FEC was able to detect the broadest spectrum of parasite species. However, FEC also missed a considerable number of infections, notably S. stercoralis, Schistosoma mansoni and G. intestinalis. PCR outperformed microscopy in terms of sensitivity and range of parasite species detected. We showed intestinal parasites-especially helminths-to be omnipresent in Inhamudima, Beira. However, it is a challenge to achieve high diagnostic sensitivity for all species. Classical techniques such as FEC are useful for the detection of some intestinal helminth species, but they lack sensitivity for other parasite species. PCR can detect intestinal parasites more accurately but is generally not feasible in resource-poor settings, at least not in peripheral labs. Hence, there is a need for a more field-friendly, sensitive approach for on-the-spot diagnosis of parasitic infections.

High Prevalence of Anaplasma spp. in Small Ruminants in Morocco.

PubMed

Ait Lbacha, H; Alali, S; Zouagui, Z; El Mamoun, L; Rhalem, A; Petit, E; Haddad, N; Gandoin, C; Boulouis, H-J; Maillard, R

2017-02-01

The prevalence of infection by Anaplasma spp. (including Anaplasma phagocytophilum) was determined using blood smear microscopy and PCR through screening of small ruminant blood samples collected from seven regions of Morocco. Co-infections of Anaplasma spp., Babesia spp, Theileria spp. and Mycoplasma spp. were investigated and risk factors for Anaplasma spp. infection assessed. A total of 422 small ruminant blood samples were randomly collected from 70 flocks. Individual animal (breed, age, tick burden and previous treatment) and flock data (GPS coordinate of farm, size of flock and livestock production system) were collected. Upon examination of blood smears, 375 blood samples (88.9%) were found to contain Anaplasma-like erythrocytic inclusion bodies. Upon screening with a large spectrum PCR targeting the Anaplasma 16S rRNA region, 303 (71%) samples were found to be positive. All 303 samples screened with the A. phagocytophilum-specific PCR, which targets the msp2 region, were found to be negative. Differences in prevalence were found to be statistically significant with regard to region, altitude, flock size, livestock production system, grazing system, presence of clinical cases and application of tick and tick-borne diseases prophylactic measures. Kappa analysis revealed a poor concordance between microscopy and PCR (k = 0.14). Agreement with PCR is improved by considering microscopy and packed cell volume (PCV) in parallel. The prevalence of double infections was found to be 1.7, 2.5 and 24% for Anaplasma-Babesia, Anaplasma-Mycoplasma and Anaplasma-Theileria, respectively. Co-infection with three or more haemoparasites was found in 1.6% of animals examined. In conclusion, we demonstrate the high burden of anaplasmosis in small ruminants in Morocco and the high prevalence of co-infections of tick-borne diseases. There is an urgent need to improve the control of this neglected group of diseases. © 2015 Blackwell Verlag GmbH.

Reliability of rapid diagnostic test for diagnosing peripheral and placental malaria in an area of unstable malaria transmission in Eastern Sudan.

PubMed

Kashif, Awadalla H; Adam, Gamal K; Mohmmed, Ahmed A; Elzaki, Salah E; AbdelHalim, Ahmed M; Adam, Ishag

2013-04-15

Diagnosing Plasmodium falciparum malaria during pregnancy is a great challenge for clinicians because of the low density of parasites in the peripheral blood and parasite sequestration in the placenta. Nevertheless, few data on the use of malaria rapid diagnostic test (RDT) during pregnancy have been published. P. falciparum infections were assessed in 156 febrile pregnant women by microscopic examination of their blood smears and by RDT and polymerase chain reactions (PCR). In addition, 150 women were assessed at the time of delivery by microscopy, RDT, PCR and placental histology investigations. The study was conducted at the Gadarif Hospital, Eastern Sudan. The SD Bioline P. f / P. v (Bio Standard Diagnostics, Gurgaon, Korea) RDT kit was evaluated in this study. Among the febrile pregnant women, 17 (11.0%), 26 (16.7%) and 18 (11.5%) positive cases of P. falciparum were detected by microscopy, RDT, and PCR, respectively. The sensitivity and specificity of the microscopy was 94.4% and 100%, respectively. The corresponding values for RDT evaluation were 83.3% and 92.0%, as compared with PCR as the gold standard.While there were no detected cases of malaria by microscopic examination of blood smears, 27 (18.0%), 21(14.0%) and 46 (30.7%) out of the 150 placentae investigated had P. falciparum as determined by RDT, PCR, and histology, respectively. The sensitivity and specificity for RDT was 17.4% and 81.7%, respectively. The corresponding values for PCR were 6.5% and 82.7%, where histology was used as the gold standard. The RDT kit used in this study has poor performance for peripheral and placental P. falciparum malaria detection in this setting. The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1092363465928479.

An Evaluation of Passive and Active Approaches to Improve Tuberculosis Notifications in Afghanistan

PubMed Central

Sanaie, A.; Nasrat, A.; Seddiq, M. K.; Mahmoodi, S. D.; Stevens, R. H.; Creswell, J.

2016-01-01

Background In Afghanistan, improving TB case detection remains challenging. In 2014, only half of the estimated incident TB cases were notified, and notifications have decreased since peaking in 2007. Active case finding has been increasingly considered to improve TB case notifications. While access to health services has improved in Afghanistan, it remains poor and many people seeking health services won’t receive proper care. Methods From October 2011 through December 2012 we conducted three separate case finding strategies in six provinces of Afghanistan and measured impact on TB case notification. Systematically screening cough among attendees at 47 health facilities, active household contact investigation of smear-positive index TB patients, and active screening at 15 camps for internally displaced people were conducted. We collected both intervention yield and official quarterly notification data. Additional TB notifications were calculated by comparing numbers of cases notified during the intervention with those notified before the intervention, then adjusting for secular trends in notification. Results We screened 2,022,127 people for TB symptoms during the intervention, tested 59,838 with smear microscopy and detected 5,046 people with smear-positive TB. Most cases (81.7%, 4,125) were identified in health facilities while nearly 20% were found through active case finding. A 56% increase in smear-positive TB notifications was observed between the baseline and intervention periods among the 47 health facilities, where cases detected by all three strategies were notified. Discussion While most people with TB are likely to be identified through health facility screening, there are many people who remain without a proper diagnosis if outreach is not attempted. This is especially true in places like Afghanistan where access to general services is poor. Targeted active case finding can improve the number of people who are detected and treated for TB and can push towards the targets of the Stop TB Global Plan and End TB Strategy. PMID:27701446

Sealer penetration into dentinal tubules in the presence or absence of smear layer: a confocal laser scanning microscopic study.

PubMed

Kuçi, Astrit; Alaçam, Tayfun; Yavaş, Ozer; Ergul-Ulger, Zeynep; Kayaoglu, Guven

2014-10-01

The aim of this study was to test the dentinal tubule penetration of AH26 (Dentsply DeTrey, Konstanz, Germany) and MTA Fillapex (Angelus, Londrina, PR, Brazil) in instrumented root canals obturated by using cold lateral compaction or warm vertical compaction techniques in either the presence or absence of the smear layer. Forty-five extracted single-rooted human mandibular premolar teeth were used. The crowns were removed, and the root canals were instrumented by using the Self-Adjusting File (ReDent-Nova, Ra'anana, Israel) with continuous sodium hypochlorite (2.6%) irrigation. Final irrigation was either with 5% EDTA or with sodium hypochlorite. The canals were dried and obturated by using rhodamine B-labeled AH26 or MTA Fillapex in combination with the cold lateral compaction or the warm vertical compaction technique. After setting, the roots were sectioned horizontally at 4-, 8-, and 12-mm distances from the apical tip. On each section, sealer penetration in the dentinal tubules was measured by using confocal laser scanning microscopy. Regardless of the usage of EDTA, MTA Fillapex, compared with AH26, was associated with greater sealer penetration when used with the cold lateral compaction technique, and, conversely, AH26, compared with MTA Fillapex, was associated with greater sealer penetration when used with the warm vertical compaction technique (P < .05). Removal of the smear layer increased the penetration depth of MTA Fillapex used with the cold lateral compaction technique (P < .05); however, it had no significant effect on the penetration depth of AH26. Greater sealer penetration could be achieved with either the MTA Fillapex-cold lateral compaction combination or with the AH26-warm vertical compaction combination. Smear layer removal was critical for the penetration of MTA Fillapex; however, the same did not hold for AH26. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

An Evaluation of Passive and Active Approaches to Improve Tuberculosis Notifications in Afghanistan.

PubMed

Sanaie, A; Mergenthaler, C; Nasrat, A; Seddiq, M K; Mahmoodi, S D; Stevens, R H; Creswell, J

2016-01-01

In Afghanistan, improving TB case detection remains challenging. In 2014, only half of the estimated incident TB cases were notified, and notifications have decreased since peaking in 2007. Active case finding has been increasingly considered to improve TB case notifications. While access to health services has improved in Afghanistan, it remains poor and many people seeking health services won't receive proper care. From October 2011 through December 2012 we conducted three separate case finding strategies in six provinces of Afghanistan and measured impact on TB case notification. Systematically screening cough among attendees at 47 health facilities, active household contact investigation of smear-positive index TB patients, and active screening at 15 camps for internally displaced people were conducted. We collected both intervention yield and official quarterly notification data. Additional TB notifications were calculated by comparing numbers of cases notified during the intervention with those notified before the intervention, then adjusting for secular trends in notification. We screened 2,022,127 people for TB symptoms during the intervention, tested 59,838 with smear microscopy and detected 5,046 people with smear-positive TB. Most cases (81.7%, 4,125) were identified in health facilities while nearly 20% were found through active case finding. A 56% increase in smear-positive TB notifications was observed between the baseline and intervention periods among the 47 health facilities, where cases detected by all three strategies were notified. While most people with TB are likely to be identified through health facility screening, there are many people who remain without a proper diagnosis if outreach is not attempted. This is especially true in places like Afghanistan where access to general services is poor. Targeted active case finding can improve the number of people who are detected and treated for TB and can push towards the targets of the Stop TB Global Plan and End TB Strategy.

A malaria diagnostic tool based on computer vision screening and visualization of Plasmodium falciparum candidate areas in digitized blood smears.

PubMed

Linder, Nina; Turkki, Riku; Walliander, Margarita; Mårtensson, Andreas; Diwan, Vinod; Rahtu, Esa; Pietikäinen, Matti; Lundin, Mikael; Lundin, Johan

2014-01-01

Microscopy is the gold standard for diagnosis of malaria, however, manual evaluation of blood films is highly dependent on skilled personnel in a time-consuming, error-prone and repetitive process. In this study we propose a method using computer vision detection and visualization of only the diagnostically most relevant sample regions in digitized blood smears. Giemsa-stained thin blood films with P. falciparum ring-stage trophozoites (n = 27) and uninfected controls (n = 20) were digitally scanned with an oil immersion objective (0.1 µm/pixel) to capture approximately 50,000 erythrocytes per sample. Parasite candidate regions were identified based on color and object size, followed by extraction of image features (local binary patterns, local contrast and Scale-invariant feature transform descriptors) used as input to a support vector machine classifier. The classifier was trained on digital slides from ten patients and validated on six samples. The diagnostic accuracy was tested on 31 samples (19 infected and 12 controls). From each digitized area of a blood smear, a panel with the 128 most probable parasite candidate regions was generated. Two expert microscopists were asked to visually inspect the panel on a tablet computer and to judge whether the patient was infected with P. falciparum. The method achieved a diagnostic sensitivity and specificity of 95% and 100% as well as 90% and 100% for the two readers respectively using the diagnostic tool. Parasitemia was separately calculated by the automated system and the correlation coefficient between manual and automated parasitemia counts was 0.97. We developed a decision support system for detecting malaria parasites using a computer vision algorithm combined with visualization of sample areas with the highest probability of malaria infection. The system provides a novel method for blood smear screening with a significantly reduced need for visual examination and has a potential to increase the throughput in malaria diagnostics.

Factors Infuencing Women in Pap Smear Uptake

NASA Astrophysics Data System (ADS)

Wijayanti, K. E.; Alam, I. G.

2017-03-01

Objective: Pap smear has proven can decrease death caused by cervical cancer. However, in Indonesia, only few woman who already did pap smear. The aim of this study was to investigate women’s knowledge about pap smear cervical cancer, and to investigate factors influence women to do pap smear test. Methods: Quantitative data colected through questionairre towards 31 women who did pap smear and 55 women who did not do pap smear. Questionairre was made using Health Belief model as a guideline to examine percieved susceptibility, perceived serioussnes, perceived benefits and perceived barriers. Chi square and multiple logistic regresion were used to investigate difference in knowledge and what the most factor that influence women to take pap smear test. Results: There’s significance knowledge difference betweeen women who did and did not do pap smear. But furthermore, by using Multiple Logistic Regression test, appearantly knowledge was not a strong predictor factor for women to take pap smear test (koefisiensi β = -0,164) Conclusion: Perceived barriers were factors that affected pap smear uptake in women in Indonesia. Few respondents get the wrong informations about pap smear, cevical cancer and its symptoms

[Multidrug-resistant tuberculosis (MDR-TB) in a black African carceral area: Experience of Mali].

PubMed

Toloba, Y; Ouattara, K; Soumaré, D; Kanouté, T; Berthé, G; Baya, B; Konaté, B; Keita, M; Diarra, B; Cissé, A; Camara, F S; Diallo, S

2018-02-01

Prison constitutes a risk factor for the emergence of multi-drug resistance of tuberculosis (MDR-TB). The aim of this work was to study MDR-TB in a black African carceral center. Prospective study from January to December 2016 at the central house of arrest for men, Bamako. The study population was composed of tuberculous detainee. The suspicion of MDR-TB was done in any tuberculosis case remained positive in the second month of first-line treatment or in contact with an MDR-TB case. Among 1622 detainee, 21 cases of pulmonary tuberculosis were notified (1.29%), with an annual incidence of 13 cases/1000 detainee, they were 16 cases of SP-PTB (microscopy smear positive tuberculosis) and five cases of microscopy smear negative tuberculosis. The mean age was 28±7 years, extremes of 18 and 46 years. A negative association was found between the notion of smoking and occupation in the occurrence of tuberculosis (OR=0.036, [95% CI: 0.03-0.04], P=0.03. Among the 21 tuberculosis cases notified, one confirmed case of MDR-TB was detected (4.7%). In the first semester of 2016 cohort, we notified a cure rate of 87.5% (7/8 SP-PTB cases), and the confirmed MDR-TB case on treatment (21-month regimen), evolution enameled of pulmonary and hearing sequelae at seven months treatment. It was the first case of MDR-TB detected in a prison in Mali. Late diagnosis, evolution is enameled of sequelae and side effects. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Wide-field computational imaging of pathology slides using lens-free on-chip microscopy.

PubMed

Greenbaum, Alon; Zhang, Yibo; Feizi, Alborz; Chung, Ping-Luen; Luo, Wei; Kandukuri, Shivani R; Ozcan, Aydogan

2014-12-17

Optical examination of microscale features in pathology slides is one of the gold standards to diagnose disease. However, the use of conventional light microscopes is partially limited owing to their relatively high cost, bulkiness of lens-based optics, small field of view (FOV), and requirements for lateral scanning and three-dimensional (3D) focus adjustment. We illustrate the performance of a computational lens-free, holographic on-chip microscope that uses the transport-of-intensity equation, multi-height iterative phase retrieval, and rotational field transformations to perform wide-FOV imaging of pathology samples with comparable image quality to a traditional transmission lens-based microscope. The holographically reconstructed image can be digitally focused at any depth within the object FOV (after image capture) without the need for mechanical focus adjustment and is also digitally corrected for artifacts arising from uncontrolled tilting and height variations between the sample and sensor planes. Using this lens-free on-chip microscope, we successfully imaged invasive carcinoma cells within human breast sections, Papanicolaou smears revealing a high-grade squamous intraepithelial lesion, and sickle cell anemia blood smears over a FOV of 20.5 mm(2). The resulting wide-field lens-free images had sufficient image resolution and contrast for clinical evaluation, as demonstrated by a pathologist's blinded diagnosis of breast cancer tissue samples, achieving an overall accuracy of ~99%. By providing high-resolution images of large-area pathology samples with 3D digital focus adjustment, lens-free on-chip microscopy can be useful in resource-limited and point-of-care settings. Copyright © 2014, American Association for the Advancement of Science.

Prevalence of abnormal Pap smears in a consecutive and previously unscreened population in Romania.

PubMed

Stolnicu, Simona; Musca, Simona; Micu, Dorian; Micu, Luminita; Moldovan, Cosmin; Puscasiu, Lucian

2014-02-01

To determine the prevalence of abnormal cervical smears in a previously unscreened and asymptomatic population in Romania and to compare the data with those from other countries in Europe. In a retrospective study, data were reviewed from smears obtained from women in Romania who had been referred to the gynecologist between January 2006 and December 2011. The smears were collected through 3 regional opportunistic programs of cervical screening and were classified according to the Bethesda system. During the study period, 50536 smear tests were carried out. Of these, 100 smears (0.2%) were unsatisfactory and excluded from the study. Among the remaining 50436 smears, 2965 patients (5.9%) had abnormal epithelial changes. Most of the abnormal smears were represented by atypical squamous cells of undetermined significance (2.6% of all smears). The data confirmed that there is a high prevalence of high-grade intraepithelial squamous-type lesions (0.9% of all smears) in Romania, and of abnormal smears in women younger than 25years of age (14.0% of all abnormal smears). The data show that there is a high prevalence of epithelial abnormalities among cervical smears in Romania compared with other European countries that run a national screening program. © 2013.

[Value of optimization of bedside Gram staining of sputum smear in the early diagnosis and treatment of ventilator-associated pneumonia].

PubMed

Liao, Xinyan; Ran, Yu; Bian, Shichang; Wang, Chao; Xu, Lei

2014-12-01

To investigate the significance of optimization of bedside Gram staining of sputum smear in the early diagnosis and antimicrobial treatment for ventilator-associated pneumonia (VAP) patients. The data of patients with VAP undergoing mechanical ventilation over 48 hours in the Department of Critical Care Medicine of Tianjin Fourth Central Hospital from June 2009 to June 2014 were analyzed. The patients were divided into two groups according to whether or not bedside Gram staining of sputum smear was used or not. The sputum samples from lower respiratory tract of all VAP patients were collected daily with tracheal catheter. In empirical examination group (from June 2009 to December 2011, n=43), the patients received antibiotics at the time of onset of VAP, selection of antibiotics depended on the information of bacterial epidemiology of the intensive care unit (ICU), and also existence of high risk factors of multi-drug resistant bacteria. In target treatment group (from January 2012 to June 2014, n=43), the patients received antibiotics according to the results of bedside instant sputum smear examination and empirical antibiotic regime. The correlation between the results of sputum smear examination and culture result was analyzed. The levels of body temperature, white blood cell (WBC) count, procalcitonin (PCT) level,and high sensitivity C- reactive protein (hs-CRP) were measured on the 1st day and 3rd day. The length of antibiotics treatment, duration of mechanical ventilation, and the time of ICU stay were recorded for both groups. There were 512 qualified sputum specimens for culture, from which 336 pathogens were found, and 358 strains of pathogenic bacteria were found from microscopic examination of 512 qualified sputum smear. The coincidence rate of results of bedside examination of sputum smear and that of sputum culture was 78.32%(401/512). The diagnostic acumen of the former was 85.42% (287/336), specificity was 64.77% (114/176), positive predictive value was 80.17% (287/358), and negative predictive value was 74.03% (114/154). On the 1st day, no statistical differences in infection index between the two groups could be found, but on the 3rd day, the results were significantly improved in both groups. Compared with the empirical treatment group, the body temperature, WBC, PCT and hs-CRP in the target treatment group were significantly lower [body temperature (centigrade): 36.83 ± 0.69 vs. 37.64 ± 0.71, WBC (× 10⁹/L): 7.91 ± 2.75 vs. 9.66 ± 3.39, PCT (μg/L): 7.14 ± 3.89 vs. 10.14 ± 4.32, hs-CRP (mg/L): 12.24 ± 6.28 vs. 15.54 ± 5.94, P<0.05 or P<0.01]. Compared with the empirical treatment group, the time of antibiotics use(days: 6.00 ± 2.55 vs. 9.20 ± 3.46), the duration of mechanical ventilation (days: 5.00 ± 1.73 vs. 7.00 ± 1.94), and the length of ICU stay (days: 7.43 ± 1.72 vs. 12.57 ± 4.16) were significantly shortened (P<0.05 or P<0.01). The results of bedside sputum examination and sputum culture showed a good correlation, and the former is helpful in early diagnosis and treatment of VAP. The result of high quality sputum smear in significant in guiding the first choice of antibiotics, reduce the time of antibiotic use, shorten the duration of mechanical ventilation and the length of ICU stay, and improve the outcome of the patients.

Optimal Design of Grid-Stiffened Composite Panels Using Global and Local Buckling Analysis

NASA Technical Reports Server (NTRS)

Ambur, Damodar R.; Jaunky, Navin; Knight, Norman F., Jr.

1996-01-01

A design strategy for optimal design of composite grid-stiffened panels subjected to global and local buckling constraints is developed using a discrete optimizer. An improved smeared stiffener theory is used for the global buckling analysis. Local buckling of skin segments is assessed using a Rayleigh-Ritz method that accounts for material anisotropy and transverse shear flexibility. The local buckling of stiffener segments is also assessed. Design variables are the axial and transverse stiffener spacing, stiffener height and thickness, skin laminate, and stiffening configuration. The design optimization process is adapted to identify the lightest-weight stiffening configuration and pattern for grid stiffened composite panels given the overall panel dimensions, design in-plane loads, material properties, and boundary conditions of the grid-stiffened panel.

Detection of isolated protein-bound metal ions by single-particle cryo-STEM.

PubMed

Elad, Nadav; Bellapadrona, Giuliano; Houben, Lothar; Sagi, Irit; Elbaum, Michael

2017-10-17

Metal ions play essential roles in many aspects of biological chemistry. Detecting their presence and location in proteins and cells is important for understanding biological function. Conventional structural methods such as X-ray crystallography and cryo-transmission electron microscopy can identify metal atoms on protein only if the protein structure is solved to atomic resolution. We demonstrate here the detection of isolated atoms of Zn and Fe on ferritin, using cryogenic annular dark-field scanning transmission electron microscopy (cryo-STEM) coupled with single-particle 3D reconstructions. Zn atoms are found in a pattern that matches precisely their location at the ferroxidase sites determined earlier by X-ray crystallography. By contrast, the Fe distribution is smeared along an arc corresponding to the proposed path from the ferroxidase sites to the mineral nucleation sites along the twofold axes. In this case the single-particle reconstruction is interpreted as a probability distribution function based on the average of individual locations. These results establish conditions for detection of isolated metal atoms in the broader context of electron cryo-microscopy and tomography.

Detection of isolated protein-bound metal ions by single-particle cryo-STEM

PubMed Central

Elad, Nadav; Bellapadrona, Giuliano; Houben, Lothar; Sagi, Irit; Elbaum, Michael

2017-01-01

Metal ions play essential roles in many aspects of biological chemistry. Detecting their presence and location in proteins and cells is important for understanding biological function. Conventional structural methods such as X-ray crystallography and cryo-transmission electron microscopy can identify metal atoms on protein only if the protein structure is solved to atomic resolution. We demonstrate here the detection of isolated atoms of Zn and Fe on ferritin, using cryogenic annular dark-field scanning transmission electron microscopy (cryo-STEM) coupled with single-particle 3D reconstructions. Zn atoms are found in a pattern that matches precisely their location at the ferroxidase sites determined earlier by X-ray crystallography. By contrast, the Fe distribution is smeared along an arc corresponding to the proposed path from the ferroxidase sites to the mineral nucleation sites along the twofold axes. In this case the single-particle reconstruction is interpreted as a probability distribution function based on the average of individual locations. These results establish conditions for detection of isolated metal atoms in the broader context of electron cryo-microscopy and tomography. PMID:28973937

Economic evaluation of DNA ploidy analysis vs liquid-based cytology for cervical screening.

PubMed

Nghiem, V T; Davies, K R; Beck, J R; Follen, M; MacAulay, C; Guillaud, M; Cantor, S B

2015-06-09

DNA ploidy analysis involves automated quantification of chromosomal aneuploidy, a potential marker of progression toward cervical carcinoma. We evaluated the cost-effectiveness of this method for cervical screening, comparing five ploidy strategies (using different numbers of aneuploid cells as cut points) with liquid-based Papanicolaou smear and no screening. A state-transition Markov model simulated the natural history of HPV infection and possible progression into cervical neoplasia in a cohort of 12-year-old females. The analysis evaluated cost in 2012 US$ and effectiveness in quality-adjusted life-years (QALYs) from a health-system perspective throughout a lifetime horizon in the US setting. We calculated incremental cost-effectiveness ratios (ICERs) to determine the best strategy. The robustness of optimal choices was examined in deterministic and probabilistic sensitivity analyses. In the base-case analysis, the ploidy 4 cell strategy was cost-effective, yielding an increase of 0.032 QALY and an ICER of $18 264/QALY compared to no screening. For most scenarios in the deterministic sensitivity analysis, the ploidy 4 cell strategy was the only cost-effective strategy. Cost-effectiveness acceptability curves showed that this strategy was more likely to be cost-effective than the Papanicolaou smear. Compared to the liquid-based Papanicolaou smear, screening with a DNA ploidy strategy appeared less costly and comparably effective.

Prevalence of K13 mutation and Day-3 positive parasitaemia in artemisinin-resistant malaria endemic area of Cambodia: a cross-sectional study.

PubMed

Kheang, Soy Ty; Sovannaroth, Siv; Ek, Sovann; Chy, Say; Chhun, Phally; Mao, Sokkieng; Nguon, Sokomar; Lek, Dy Soley; Menard, Didier; Kak, Neeraj

2017-09-13

The presence of artemisinin-resistant malaria parasites was confirmed in western Cambodia in 2009. In 2013, mutations in the propeller domain of the kelch protein K13 was found to be associated with artemisinin resistance. A cross-sectional study was conducted to determine the prevalence of Day-3 parasitaemia, estimate the frequency of k13 molecular marker and assess their relationship in the context of operational research. Blood smears and filter paper blood spots were collected from febrile patients in Kravanh District, Pursat Province. The blood smears were examined by microscopy, and blood spots by a k13 mutation assay. Data from 92 patients were analysed. Only one was positive for Day-3 parasitaemia. Results of the k13 assay were interpretable for 76 of the 92 samples. The findings were: wild type: 9 (12%), C580Y: 64 (84%), Y493H: 3 (4%). Therefore, despite the high prevalence of k13 mutants (67/76: 88%), only 1 of the 92 patients remained blood smear positive for Plasmodium falciparum on Day-3. These preliminary findings suggest good potency of artemisinin despite the dominance of k13 mutation in Kravanh, but the result is not necessarily representative of the western part of Cambodia. Further investigation should be made to determine if k13 marker remains useful as a tool for tracking artemisinin resistance and predicting the trend of the efficacy of artemisinin combination therapy once the mutant alleles have been well established in the population.

Cytologic features of the normal pineal gland on squash preparations.

PubMed

Murro, Diana; Alsadi, Alaa; Nag, Sukriti; Arvanitis, Leonidas; Gattuso, Paolo

2014-11-01

As primary pineal lesions are extremely rare, many surgical pathologists are unfamiliar with normal pineal cytologic features. We describe cytologic features of the normal pineal gland in patients of varying ages and identify common diagnostic pitfalls. We performed a retrospective review of pineal gland biopsies performed at our institution, where approximately 30,000 surgical specimens are accessioned yearly, for the last 23 years. Only two pineal gland biopsies were found. Although both cases were initially diagnosed as low-grade gliomas on frozen section, the final diagnosis was benign pineal tissue based on light microscopy and immunohistochemistry results. Additionally, we performed squash preparations of five normal pineal gland autopsy specimens with Papanicolaou and Diff-Quik® (Dade Behring, Newark, DE) stains. Infant preparations were highly cellular smears composed of numerous, uniform, single cells with indistinct cytoplasm, small round-to-oval nuclei, fine chromatin, and absent nucleoli and calcifications. The vague microfollicular pattern mimicked a pineocytoma and the fine fibrillary background mimicked a glial neoplasm. Young adult smears were similar; however, microcalcifications were present with fewer background single cells. Older patients had much less cellular smears composed of small clusters of cells with fusiform-to-spindle nuclei, a fine chromatin pattern, and indistinct cytoplasmic borders. There were fewer background single cells and more microcalcifications. The cytologic features of the native pineal gland vary with age. Normal pineal tissue can be confused with a pineocytoma or low-grade glioma. Familiarity with normal pineal gland cytological features will help to avoid a potential misdiagnosis. © 2014 Wiley Periodicals, Inc.

Plastic spatula with narrow long tip provides higher satisfactory smears for Pap test.

PubMed

Kaur, Pervinder; Kushtagi, Pralhad

2013-07-01

Ayre spatula for cervical smear collection is being used despite the suggestion that different modified spatulas provide more satisfactory sampling. To see whether the cytological pickup improves with the use of long tipped spatula. Rurally based University Hospital; crossover study. Pap smear using Ayre spatula in 500 and with plastic narrow long tip (Szalay) spatula in 500 clinic attending women was taken and analyzed. Crossover smears were taken with modified spatula in 163 and using Ayre spatula in 187 women after 2 weeks of initial smears. The same pathologist made cytological reporting for all smears and was unaware of the type of spatula used. Smears from Ayre spatula had significantly higher reports of inadequate smears (94 of 500 vs. 68 of 500 for Ayre and Szalay, respectively; P = 0.032) and it remained so even after crossover (94 of 187 vs. 70 of 163 for Ayre and Szalay, respectively; P = 0.2). Cellular quality appeared better with smears taken using Szalay spatula, but the overall abnormal smear detection rate remained similar with either collection tool (χ(2) = 1.5; P = 0.2). Proportion of satisfactory smears is higher when long tip plastic spatula is used for collection of sample.

Young Singaporean women's knowledge of cervical cancer and pap smear screening: a descriptive study.

PubMed

Shea, Juanna; Klainin-Yobas, Piyanee; Mackey, Sandra

2013-12-01

To assess the knowledge of young female Singaporeans regarding cervical cancer and pap smear, the intention to participate in pap smear and whether there is any relationship between knowledge and intention to participate in pap smear screening. While cervical cancer has poor prognosis in the later stages, pap smear is effective in identifying precancerous lesions, which are more treatable. Pap smear screening is available to women in Singapore, but its uptake is opportunistic. Research has shown that knowledge about pap smear and cervical cancer is important determinant of screening behaviour in Singaporean women. Cross-sectional descriptive correlational design was used. Three hundred and ninety-three young Singaporean undergraduates, aged 18-25 years, were recruited via convenience sampling from a local university over a four-month period. Data were collected through self-administered questionnaires. Majority of the participants knew the term 'pap smear' and its function. However, knowledge of the risk factors for cervical cancer was lacking among the young women. Knowledge of pap smear and cervical cancer had a weak correlation with the intention to go for the future uptake of pap smear. Educational efforts among younger Singaporean women on the knowledge of pap smear and risk factors for cervical cancer are needed. Improving knowledge will enable them to understand the importance of reducing exposure to risk factors and regular pap smear screening. All health professionals working with young Asian women should be prepared to educate and counsel young women to participate in pap smear screening according to current guidelines. In particular, knowledge of the age to attend the first pap smear and the recommended frequency for screening need to be targeted for health education. © 2013 John Wiley & Sons Ltd.

Epitope enhancement for immunohistochemical demonstration of tartrate-resistant acid phosphatase.

PubMed

Janckila, A J; Lear, S C; Martin, A W; Yam, L T

1996-03-01

We have developed a monoclonal antibody (9C5) for immunohistochemical localization of tartrate-resistant acid phosphatase (TRAcP). This antibody reacts with a denatured epitope of TRAcP and requires enhancement methods to promote antigenicity in paraffin-embedded tissues. We used this antibody to systematically examine proteolytic digestion and heat denaturation conditions for epitope enhancement in both paraffin sections and fixed smears. The goal was to increase the sensitivity of the immunohistochemical stain for TRAcP. Optimal conditions for proteolytic digestion were established. Denaturation in a conventional boiling water bath was compared to microwave irradiation in several commonly used solutions. Immunohistochemistry was compared directly to TRAcP cytochemistry in fixed smears from hairy cell leukemia specimens to gauge the level of sensitivity of our improved method. Attempts were made to "retrieve" the 9C5 epitope from overfixed tissues and aged smears. Maximal immunoreactivity of TRAcP was achieved by microwave irradiation in a citrate or Tris buffer of pH 6.0-8.0 without the need for a subsequent protease digestion step. With this method of epitope enhancement, immunohistochemistry with antibody 9C5 was as sensitive as direct cytochemical staining of TRAcP activity. However, once a tissue specimen had been overfixed or a smear stored for a year or more, the 9C5 epitope was no longer retrievable. The key element in epitope enhancement for 9C5 immunohistochemistry is heat denaturation of the target epitope. Immunohistochemistry of TRAcP in paraffin sections would be a great asset to the study of specialized forms of the monocyte/macrophage lineage and to the process of macrophage activation. It would also provide another means for more precise evaluation of residual disease in bone marrow of patients treated for hairy cell leukemia.

Prevalence of pulmonary TB and spoligotype pattern of Mycobacterium tuberculosis among TB suspects in a rural community in Southwest Ethiopia

PubMed Central

2012-01-01

Background In Ethiopia where there is no strong surveillance system and state of the art diagnostic facilities are limited, the real burden of tuberculosis (TB) is not well known. We conducted a community based survey to estimate the prevalence of pulmonary TB and spoligotype pattern of the Mycobacterium tuberculosis isolates in Southwest Ethiopia. Methods A total of 30040 adults in 10882 households were screened for pulmonary TB in Gilgel Gibe field research centre in Southwest Ethiopia. A total of 482 TB suspects were identified and smear microscopy and culture was done for 428 TB suspects. Counseling and testing for HIV/AIDS was done for all TB suspects. Spoligotyping was done to characterize the Mycobacterium tuberculosis isolates. Results Majority of the TB suspects were females (60.7%) and non-literates (83.6%). Using smear microscopy, a total of 5 new and 4 old cases of pulmonary TB cases were identified making the prevalence of TB 30 per 100,000. However, using the culture method, we identified 17 new cases with a prevalence of 76.1 per 100,000. There were 4.3 undiagnosed pulmonary TB cases for every TB case who was diagnosed through the passive case detection mechanism in the health facility. Eleven isolates (64.7%) belonged to the six previously known spoligotypes: T, Haarlem and Central-Asian (CAS). Six new spoligotype patterns of Mycobacterium tuberculosis, not present in the international database (SpolDB4) were identified. None of the rural residents was HIV infected and only 5 (5.5%) of the urban TB suspects were positive for HIV. Conclusion The prevalence of TB in the rural community of Southwest Ethiopia is low. There are large numbers of undiagnosed TB cases in the community. However, the number of sputum smear-positive cases was very low and therefore the risk of transmitting the infection to others may be limited. Active case finding through health extension workers in the community can improve the low case detection rate in Ethiopia. A large scale study on the genotyping of Mycobacterium tuberculosis in Ethiopia is crucial to understand transmission dynamics, identification of drug resistant strains and design preventive strategies. PMID:22414165

The incidence and clinical implication of sputum with positive acid-fast bacilli smear but negative in mycobacterial culture in a tertiary referral hospital in South Korea.

PubMed

Lee, Jae Seok; Kim, Eui-Chong; Joo, Sei Ick; Lee, Sang-Min; Yoo, Chul-Gyu; Kim, Young Whan; Han, Sung Koo; Shim, Young-Soo; Yim, Jae-Joon

2008-10-01

Although it is not rare to find sputum that is positive acid-fast bacilli (AFB) smear but subsequent culture fails to isolate mycobacteria in clinical practice, the incidence and clinical implication of those sputa from new patients has not been clearly elucidated. The aim of this study was to determine the incidence and clinical implication of sputum with positive AFB smear but negative in mycobacterial culture. All sputa that were positive AFB smear requested during diagnostic work up for new patients visiting Seoul National University Hospital from 1 January 2005 through 31 December 2006 were included. Sputa producing a positive AFB smear but negative mycobacterial culture were classified into one of four categories: laboratory failure to isolate mycobacteria, false positive AFB smear, pathogen may show a positive AFB smear other than mycobacteria, and indeterminate results. Out of 447 sputa with a positive AFB smear, 29 (6.5%) failed to culture any organism. Among these 29 sputa, 18 were caused by laboratory failure to isolate mycobacteria, six were false positive smears, and five indeterminate. Although most sputum with a positive AFB smear but negative culture could be classified as a laboratory failure, clinicians should consider the possibility of false positive AFB smear.

Factors associated with smoking among tuberculosis patients in Spain.

PubMed

Jiménez-Fuentes, María Ángeles; Rodrigo, Teresa; Altet, María Neus; Jiménez-Ruiz, Carlos A; Casals, Martí; Penas, Antón; Mir, Isabel; Solano Reina, Segismundo; Riesco-Miranda, Juan Antonio; Caylá, Joan A

2016-09-14

To determine the prevalence of smoking and analyze associated factors in a cohort of patients diagnosed with tuberculosis (TB) in Spain between 2006 and 2013. Multicenter, cross-sectional, descriptive, observational study using a national database of TB patients, using logistic regression to calculate odds ratios (OR) and confidence intervals (CI). We analyzed 5,846 cases (62 % men, mean age 39 years, 33 % foreigners). 23.4 % were alcohol abuser, 1.3 % were injected drug users (IDU), 4.6 % were co-infected with HIV, and 7.5 % had a history of TB treatment. 6.6 % and 0.8 % showed resistance to one and multiple drugs, respectively. The predominant clinical presentation was pulmonary (71 %) with a cavitary radiological pattern in 32.8 % of cases. 82 % of cases were confirmed microbiologically, and 54 % were smear-positive microscopy. 2,300 (39.3 %) patients were smokers. The following factors were associated with smoking: male sex (OR = 2.26;CI:1.97;2.60), Spanish origin (OR = 2.79;CI:2.40-3.24), alcoholism (OR = 2.85;CI:2.46;3.31), IDU (OR = 2.78;CI:1.48;5.52), homelessness (OR = 1.99;CI:1.14-3.57), pulmonary TB (OR = 1.61;CI:1.16;2.24), cavitary radiological pattern (OR = 1.99;CI:1.43;2.79) and a smear-positive microscopy at the time of diagnosis (OR = 1.39;CI:1.14;1.17). The prevalence of smoking among TB patients is high. Smokers with TB have a distinct sociodemographic, clinical, radiological and microbiological profile to non-smokers.

Cutting efficiency of a mid-infrared laser on human enamel.

PubMed

Levy, G; Koubi, G F; Miserendino, L J

1998-02-01

In this study, the cutting ability of a newly developed dental laser was compared with a dental high-speed handpiece and rotary bur for removal of enamel. Measurements of the volume of tissue removed, energy emitted, and time of exposure were used to quantify the ablation rate (rate of tissue removal) for each test group and compared. Cutting efficiency (mm3/s) of the laser was calculated based on the mean volume of tissue removed per pulse (mm3/pulse) and unit energy expended (mm3/J) over the range of applied powers (2, 4, 6, and 8 W). The specimens were then examined by light microscopy and scanning electron micrographs for qualitative analysis of the amount of remaining debris and the presence of the smear layer on the prepared enamel surface. Calculations of the cutting efficiency of the laser over the range of powers tested revealed a linear relationship with the level of applied power. The maximum average rate of tissue removal by the laser was 0.256 mm3/s at 8 W, compared with 0.945 mm3/s by the dental handpiece. Light microscopy and scanning electron micrograph examinations revealed a reduction in the amount of remaining debris and smear layer in the laser-prepared enamel surfaces, compared with the conventional method. Based on the results of this study, the cutting efficiency of the high-speed handpiece and dental bur was 3.7 times greater than the laser over the range of powers tested, but the laser appeared to create a cleaner enamel surface with minimal thermal damage. Further modifications of the laser system are suggested for improvement of laser cutting efficiency.

Bacteriologic monitoring of multidrug-resistant tuberculosis patients in five DOTS-Plus pilot projects.

PubMed

Gammino, V M; Taylor, A B; Rich, M L; Bayona, J; Becerra, M C; Bonilla, C; Gelmanova, I; Hollo, V; Jaramillo, E; Keshavjee, S; Leimane, V; Mitnick, C D; Quelapio, M I D; Riektsina, V; Tupasi, T E; Wells, C D; Zignol, M; Cegielski, P J

2011-10-01

Multidrug-resistant tuberculosis programs in DOTS-Plus pilot sites in five countries. To calculate sputum conversion time and its relationship to treatment outcome, document the frequency of culture reversions and examine concordance of smear and culture to assess the potential consequences of monitoring by smear microscopy alone. Retrospective cohort analysis of 1926 patients receiving individualized, second-line therapy. Among 1385 sputum culture-positive cases at baseline, 1146 (83%) experienced at least one culture conversion during treatment. Conversion, however, was not sustained in all patients: 201 (15%) experienced initial culture conversion and at least one subsequent culture reversion to positive; 1064 (77%) achieved sustained culture conversion. Median time to culture conversion was 3 months. Among 206 patients whose nal conversion occurred 7-18 months after the initiation of therapy, 71% were cured or had completed treatment. Prolonged treatment for patients with delayed conversion may be beneficial, as 71% of late converters still achieved cure or completed treatment. This has implications for programs with de ned end points for treatment failure. The interval between rst and nal conversion among patients whose initial con- version is not sustained raises concern with respect to the ongoing debate regarding duration of treatment and the definition of cure.

Comparison of scanty AFB smears against culture in an area with high HIV prevalence.

PubMed

Lawson, L; Yassin, M A; Ramsay, A; Emenyonu, N E; Squire, S B; Cuevas, L E

2005-08-01

To verify among tuberculosis (TB) suspects attending hospitals in Abuja, Nigeria, if sputum smears graded as scanty are false-positive, sputum smears from 1068 patients were graded with the International Union Against Tuberculosis and Lung Disease classification. One specimen was cultured. Eight hundred and twenty-four (26%) smears were positive, 137 (4%) scanty and 2243 negative. Of 1068 cultures, 680 (64%) were positive. One hundred and thirty (95%) scanty and 809 (98%) positive smears were culture-positive. Twelve of 18 patients with a single scanty smear and 51 of 52 with > or = 2 scanty smears were culture-positive. Fewer than < 5% scanty results, < 1% of the patients treated for TB, are false-positive.

Maximizing the Biochemical Resolving Power of Fluorescence Microscopy

PubMed Central

Esposito, Alessandro; Popleteeva, Marina; Venkitaraman, Ashok R.

2013-01-01

Most recent advances in fluorescence microscopy have focused on achieving spatial resolutions below the diffraction limit. However, the inherent capability of fluorescence microscopy to non-invasively resolve different biochemical or physical environments in biological samples has not yet been formally described, because an adequate and general theoretical framework is lacking. Here, we develop a mathematical characterization of the biochemical resolution in fluorescence detection with Fisher information analysis. To improve the precision and the resolution of quantitative imaging methods, we demonstrate strategies for the optimization of fluorescence lifetime, fluorescence anisotropy and hyperspectral detection, as well as different multi-dimensional techniques. We describe optimized imaging protocols, provide optimization algorithms and describe precision and resolving power in biochemical imaging thanks to the analysis of the general properties of Fisher information in fluorescence detection. These strategies enable the optimal use of the information content available within the limited photon-budget typically available in fluorescence microscopy. This theoretical foundation leads to a generalized strategy for the optimization of multi-dimensional optical detection, and demonstrates how the parallel detection of all properties of fluorescence can maximize the biochemical resolving power of fluorescence microscopy, an approach we term Hyper Dimensional Imaging Microscopy (HDIM). Our work provides a theoretical framework for the description of the biochemical resolution in fluorescence microscopy, irrespective of spatial resolution, and for the development of a new class of microscopes that exploit multi-parametric detection systems. PMID:24204821

Automatic diagnosis of malaria based on complete circle-ellipse fitting search algorithm.

PubMed

Sheikhhosseini, M; Rabbani, H; Zekri, M; Talebi, A

2013-12-01

Diagnosis of malaria parasitemia from blood smears is a subjective and time-consuming task for pathologists. The automatic diagnostic process will reduce the diagnostic time. Also, it can be worked as a second opinion for pathologists and may be useful in malaria screening. This study presents an automatic method for malaria diagnosis from thin blood smears. According to this fact that malaria life cycle is started by forming a ring around the parasite nucleus, the proposed approach is mainly based on curve fitting to detect parasite ring in the blood smear. The method is composed of six main phases: stain object extraction step, which extracts candidate objects that may be infected by malaria parasites. This phase includes stained pixel extraction step based on intensity and colour, and stained object segmentation by defining stained circle matching. Second step is preprocessing phase which makes use of nonlinear diffusion filtering. The process continues with detection of parasite nucleus from resulted image of previous step according to image intensity. Fourth step introduces a complete search process in which the circle search step identifies the direction and initial points for direct least-square ellipse fitting algorithm. Furthermore in the ellipse searching process, although parasite shape is completed undesired regions with high error value are removed and ellipse parameters are modified. Features are extracted from the parasite candidate region instead of whole candidate object in the fifth step. By employing this special feature extraction way, which is provided by special searching process, the necessity of employing clump splitting methods is removed. Also, defining stained circle matching process in the first step speeds up the whole procedure. Finally, a series of decision rules are applied on the extracted features to decide on the positivity or negativity of malaria parasite presence. The algorithm is applied on 26 digital images which are provided from thin blood smear films. The images are contained 1274 objects which may be infected by parasite or healthy. Applying the automatic identification of malaria on provided database showed a sensitivity of 82.28% and specificity of 98.02%. © 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

[Effects of Nd: YAG laser irradiation on the root surfaces and adhesion of Streptococcus mutans].

PubMed

Yuanhong, Li; Zhongcheng, Li; Mengqi, Luo; Daonan, Shen; Shu, Zhang; Shu, Meng

2016-12-01

This study aimed to evaluate the effects of treatment with different powers of Nd: YAG laser irradiation on root surfaces and Streptococcus mutans (S. mutans) adhesion. Extracted teeth because of severe periodontal disease were divided into the following four groups: control group, laser group 1, laser group 2, and laser group 3. After scaling and root planning, laser group 1, laser group 2, and laser group 3 were separately treated with Nd: YAG laser irradiation (4/6/8 W, 60 s); however, the control group did not receive the treatment. Scanning electron microscopy (SEM) was used to determine the morphology. S. mutans were cultured with root slices from each group. Colony forming unit per mL (CFU·mL⁻¹) was used to count and compare the amounts of bacteria adhesion among groups. SEM was used to observe the difference of bacteria adhesion to root surfaces between control group (scaling) and laser group 2 (6 W, 60 s), thereby indicating the different bacteria adhesions because of different treatments. Morphology alterations indicated that root surfaces in control group contain obvious smear layer, debris, and biofilm; whereas the root surfaces in laser group contain more cracks with less smear layer and debris. The bacteria counting indicated that S. mutans adhesion to laser group was weaker than that of control group (P<0.05). No statistical significance among the laser groups (P>0.05) was observed. Morphology alterations also verified that S. mutans adhesion to laser group 2 (6 W, 60 s) was weaker than that of control group (scaling). This study demonstrated that Nd: YAG laser irradiation treatment after scaling can reduce smear layer, debris, and biofilm on the root surfaces as compared with conventional scaling. The laser treatment reduces the adhesion of S. mutans as well. However, Nd: YAG laser irradiation can cause cracks on the root surfaces. In this experiment, the optimum laser power of 6 W can thoroughly remove the smear layer and debris, as well as relatively improve the control of thermal damagee.

Catching the missing million: experiences in enhancing TB & DR-TB detection by providing upfront Xpert MTB/RIF testing for people living with HIV in India.

PubMed

Raizada, Neeraj; Sachdeva, Kuldeep Singh; Sreenivas, Achuthan; Kulsange, Shubhangi; Gupta, Radhey Shyam; Thakur, Rahul; Dewan, Puneet; Boehme, Catharina; Paramsivan, Chinnambedu Nainarappan

2015-01-01

A critical challenge in providing TB care to People Living with HIV (PLHIV) is establishing an accurate bacteriological diagnosis. Xpert MTB/RIF, a highly sensitive and specific rapid tool, offers a promising solution in addressing these challenges. This study presents results from PLHIV taking part in a large demonstration study across India wherein upfront Xpert MTB/RIF testing was offered to all presumptive PTB cases in public health facilities. The study covered a population of 8.8 million across 18 sub-district level tuberculosis units (TU), with one Xpert MTB/RIF platform established at each TU. All HIV-infected patients suspected of TB (both TB and Drug Resistant TB (DR-TB)) accessing public health facilities in study area were prospectively enrolled and provided upfront Xpert MTB/RIF testing. 2,787 HIV-infected presumptive pulmonary TB cases were enrolled and 867 (31.1%, 95% Confidence Interval (CI) 29.4‒32.8) HIV-infected TB cases were diagnosed under the study. Overall 27.6% (CI 25.9-29.3) of HIV-infected presumptive PTB cases were positive by Xpert MTB/RIF, compared with 12.9% (CI 11.6-14.1) who had positive sputum smears. Upfront Xpert MTB/RIF testing of presumptive PTB and DR-TB cases resulted in diagnosis of 73 (9.5%, CI 7.6‒11.8) and 16 (11.2%, CI 6.7‒17.1) rifampicin resistance cases, respectively. Positive predictive value (PPV) for rifampicin resistance detection was high 97.7% (CI 89.3‒99.8), with no significant difference with or without prior history of TB treatment. The study results strongly demonstrate limitations of using smear microscopy for TB diagnosis in PLHIV, leading to low TB and DR-TB detection which can potentially lead to either delayed or sub-optimal TB treatment. Our findings demonstrate the usefulness and feasibility of addressing this diagnostic gap with upfront of Xpert MTB/RIF testing, leading to overall strengthening of care and support package for PLHIV.

Catching the Missing Million: Experiences in Enhancing TB & DR-TB Detection by Providing Upfront Xpert MTB/RIF Testing for People Living with HIV in India

PubMed Central

Raizada, Neeraj; Sachdeva, Kuldeep Singh; Sreenivas, Achuthan; Kulsange, Shubhangi; Gupta, Radhey Shyam; Thakur, Rahul; Dewan, Puneet; Boehme, Catharina; Paramsivan, Chinnambedu Nainarappan

2015-01-01

Background A critical challenge in providing TB care to People Living with HIV (PLHIV) is establishing an accurate bacteriological diagnosis. Xpert MTB/RIF, a highly sensitive and specific rapid tool, offers a promising solution in addressing these challenges. This study presents results from PLHIV taking part in a large demonstration study across India wherein upfront Xpert MTB/RIF testing was offered to all presumptive PTB cases in public health facilities. Method The study covered a population of 8.8 million across 18 sub-district level tuberculosis units (TU), with one Xpert MTB/RIF platform established at each TU. All HIV-infected patients suspected of TB (both TB and Drug Resistant TB (DR-TB)) accessing public health facilities in study area were prospectively enrolled and provided upfront Xpert MTB/RIF testing. Result 2,787 HIV-infected presumptive pulmonary TB cases were enrolled and 867 (31.1%, 95% Confidence Interval (CI) 29.4‒32.8) HIV-infected TB cases were diagnosed under the study. Overall 27.6% (CI 25.9–29.3) of HIV-infected presumptive PTB cases were positive by Xpert MTB/RIF, compared with 12.9% (CI 11.6–14.1) who had positive sputum smears. Upfront Xpert MTB/RIF testing of presumptive PTB and DR-TB cases resulted in diagnosis of 73 (9.5%, CI 7.6‒11.8) and 16 (11.2%, CI 6.7‒17.1) rifampicin resistance cases, respectively. Positive predictive value (PPV) for rifampicin resistance detection was high 97.7% (CI 89.3‒99.8), with no significant difference with or without prior history of TB treatment. Conclusion The study results strongly demonstrate limitations of using smear microscopy for TB diagnosis in PLHIV, leading to low TB and DR-TB detection which can potentially lead to either delayed or sub-optimal TB treatment. Our findings demonstrate the usefulness and feasibility of addressing this diagnostic gap with upfront of Xpert MTB/RIF testing, leading to overall strengthening of care and support package for PLHIV. PMID:25658091

Who is getting Pap smears in urban Peru?

PubMed Central

Paz Soldan, Valerie A; Lee, Frank H; Carcamo, Cesar; Holmes, King K; Garnett, Geoff P; Garcia, Patricia

2008-01-01

Background Cervical cancer, although usually preventable by Pap smear screening, remains the leading cause of cancer-related deaths among women in Peru. The percentages and characteristics of women in Peru who have or have not had a Pap smear have not been defined. Methods In an urban community randomized trial of sexually transmitted infection (STI)/HIV prevention in Peru, 6712 randomly selected women between the ages of 18 and 29 from 20 cities were interviewed regarding having had cervical Pap smears. Results Among women sampled, only 30.9% had had a Pap smear. By multivariate analysis, the main predictors of having a Pap smear were having had sex, having had children, completion of secondary education and increasing age. Regional variations were also found: women from the highlands and rainforest were less likely to have had Pap smears than women from the coast. Conclusion A norm of seeking and receiving Pap smears has not been established among sexually active young Peruvian women. To improve Pap smear coverage in Peru, promotion efforts should target underserved women and regions with less coverage. PMID:18653515

Quality of diagnosis and monitoring of tuberculosis in Northern Ethiopia: medical records-based retrospective study.

PubMed

Mala, George; Spigt, Mark G; Gidding, Luc G; Blanco, Roman; Dinant, Geert-Jan

2015-10-01

To determine quality of diagnosis and monitoring of treatment response of patients with smear-negative pulmonary tuberculosis (TB) compared with smear-positive cases in Ethiopia. A retrospective analysis of medical records of newly diagnosed pulmonary TB cases that were registered for taking anti-TB medication and had completed treatment between 2010 and 2012. We evaluated the percentage of cases that were managed according to the International Standards of Tuberculosis Care (ISTC) and compared smear-negative with smear-positive cases. We analysed 1168 cases of which 742 (64%) were sputum smear-negative cases. Chest radiography examination at diagnosis and microbiological testing at the end of the intensive phase of treatment was performed in a smaller proportion than in smear-positive TB cases (70% vs. 79%, P value <0.001) and (70% vs. 95%, P value <0.001), respectively. Clinical actions recommended in the ISTC are of greatest importance in minimising pitfalls in care of smear-negative TB yet were performed less often in smear-negative than smear-positive TB cases. © The Author(s) 2015.

White blood cell counting analysis of blood smear images using various segmentation strategies

NASA Astrophysics Data System (ADS)

Safuan, Syadia Nabilah Mohd; Tomari, Razali; Zakaria, Wan Nurshazwani Wan; Othman, Nurmiza

2017-09-01

In white blood cell (WBC) diagnosis, the most crucial measurement parameter is the WBC counting. Such information is widely used to evaluate the effectiveness of cancer therapy and to diagnose several hidden infection within human body. The current practice of manual WBC counting is laborious and a very subjective assessment which leads to the invention of computer aided system (CAS) with rigorous image processing solution. In the CAS counting work, segmentation is the crucial step to ensure the accuracy of the counted cell. The optimal segmentation strategy that can work under various blood smeared image acquisition conditions is remain a great challenge. In this paper, a comparison between different segmentation methods based on color space analysis to get the best counting outcome is elaborated. Initially, color space correction is applied to the original blood smeared image to standardize the image color intensity level. Next, white blood cell segmentation is performed by using combination of several color analysis subtraction which are RGB, CMYK and HSV, and Otsu thresholding. Noises and unwanted regions that present after the segmentation process is eliminated by applying a combination of morphological and Connected Component Labelling (CCL) filter. Eventually, Circle Hough Transform (CHT) method is applied to the segmented image to estimate the number of WBC including the one under the clump region. From the experiment, it is found that G-S yields the best performance.

Tuberculosis screening in patients with HIV: use of audit and feedback to improve quality of care in Ghana

PubMed Central

Bjerrum, Stephanie; Bonsu, Frank; Hanson-Nortey, Nii Nortey; Kenu, Ernest; Johansen, Isik Somuncu; Andersen, Aase Bengaard; Bjerrum, Lars; Jarbøl, Dorte; Munck, Anders

2016-01-01

Background Tuberculosis screening of people living with HIV (PLHIV) can contribute to early tuberculosis diagnosis and improved patient outcomes. Evidence-based guidelines for tuberculosis screening are available, but literature assessing their implementation and the quality of clinical practice is scarce. Objectives To assess tuberculosis screening practices and the effectiveness of audit and performance feedback to improve quality of tuberculosis screening at HIV care clinics in Ghana. Design Healthcare providers at 10 large HIV care clinics prospectively registered patient consultations during May and October 2014, before and after a performance feedback intervention in August 2014. The outcomes of interest were overall tuberculosis suspicion rate during consultations and provider adherence to the International Standards for Tuberculosis Care and the World Health Organizations’ guidelines for symptom-based tuberculosis screening among PLHIV. Results Twenty-one healthcare providers registered a total of 2,666 consultations; 1,368 consultations before and 1,298 consultations after the feedback intervention. Tuberculosis suspicion rate during consultation increased from 12.6 to 20.9% after feedback (odds ratio, OR 1.83; 95% confidence interval, CI: 1.09–3.09). Before feedback, sputum smear microscopy was requested for 58.7% of patients with suspected tuberculosis, for 47.2% of patients with cough ≥2 weeks, and for 27.5% of patients with a positive World Health Organization (WHO) symptom screen (any of current cough, fever, weight loss or night sweats). After feedback, patients with a positive WHO symptom screen were more likely to be suspected of tuberculosis (OR 2.21; 95% CI: 1.19–4.09) and referred for microscopy (OR 2.71; 95% CI: 1.25–5.86). Conclusions A simple prospective audit tool identified flaws in clinical practices for tuberculosis screening of PLHIV. There was no systematic identification of people with suspected active tuberculosis. We found low initial tuberculosis suspicion rate compounded by low referral rates of relevant patients for sputum smear microscopy. Adherence to recommended standards and guidelines for tuberculosis screening improved after performance feedback. PMID:27569593

Evaluating the impact of decentralising tuberculosis microscopy services to rural township hospitals in gansu province, china

PubMed Central

2011-01-01

Background In 2004, the Ministry of Health issued the policy of decentralising microscopy services (MCs) to one third of all township hospitals in China. The study was conducted in Gansu Province, a poor western one in China. Ganzhou was one county in Gansu Province. Ganzhou County was identified as a unique case of further decentralisation of tuberculosis (TB) treatment services in township hospitals. The study evaluated the impact of the MC policy on providers and patients in Gansu Province. The second objective was to assess the unique case of Ganzhou County compared with other counties in the province. Methods Both quantitative and qualitative methods were used. All 523 MCs in the province completed an institutional survey regarding their performance. Four counties were selected for in-depth investigation, where 169 TB suspects were randomly selected from the MC and county TB dispensary registers for questionnaire surveys. Informant interviews were conducted with 38 health staff at the township and county levels in the four counties. Results Gansu established MCs in 39% of its township hospitals. From January 2006 to June 2007, 8% of MCs identified more than 10 TB sputum smear positive patients while 54% did not find any. MCs identified 1546 TB sputum smear positive patients, accounting for 9% of the total in the province. The throughputs of MCs in Ganzhou County were eight times of those in other counties. Interviews identified several barriers to implement the MC policy, such as inadequate health financing, low laboratory capacity, lack of human resources, poor treatment and management capacities, and lack of supervisions from county TB dispensaries. Conclusion Microscopy centre throughputs were generally low in Gansu Province, and the contribution of MCs to TB case detection was insignificant taking account the number of MCs established. As a unique case of full decentralisation of TB service, Ganzhou County presented better results. However, standards and quality of TB care needed to be improved. The MC policy needs to be reviewed in light of evidence from this study. PMID:21324106

Diagnosing Polyparasitism in a High-Prevalence Setting in Beira, Mozambique: Detection of Intestinal Parasites in Fecal Samples by Microscopy and Real-Time PCR

PubMed Central

Polderman, Anton M.; Vinkeles Melchers, Natalie V. S.; Brienen, Eric A. T.; Verweij, Jaco J.; Groosjohan, Bernhard; Mendes, Felisberto; Mechendura, Manito; Hepp, Dagmar H.; Langenberg, Marijke C. C.; Edelenbosch, Rosanne; Polman, Katja; van Lieshout, Lisette

2017-01-01

Background Many different intestinal parasite species can co-occur in the same population. However, classic diagnostic tools can only frame a particular group of intestinal parasite species. Hence, one or two tests do not suffice to provide a complete picture of infecting parasite species in a given population. The present study investigated intestinal parasitic infections in Beira, Mozambique, i.e. in the informal settlement of Inhamudima. Diagnostic accuracy of five classical microscopy techniques and real-time PCR for the detection of a broad spectrum of parasites was compared. Methodology/Principal Findings A cross-sectional population-based survey was performed. One stool sample per participant (n = 303) was examined by direct smear, formal-ether concentration (FEC), Kato smear, Baermann method, coproculture and real-time PCR. We found that virtually all people (96%) harbored at least one helminth, and that almost half (49%) harbored three helminths or more. Remarkably, Strongyloides stercoralis infections were widespread with a prevalence of 48%, and Ancylostoma spp. prevalence was higher than that of Necator americanus (25% versus 15%), the hookworm species that is often assumed to prevail in East-Africa. Among the microscopic techniques, FEC was able to detect the broadest spectrum of parasite species. However, FEC also missed a considerable number of infections, notably S. stercoralis, Schistosoma mansoni and G. intestinalis. PCR outperformed microscopy in terms of sensitivity and range of parasite species detected. Conclusions/Significance We showed intestinal parasites—especially helminths—to be omnipresent in Inhamudima, Beira. However, it is a challenge to achieve high diagnostic sensitivity for all species. Classical techniques such as FEC are useful for the detection of some intestinal helminth species, but they lack sensitivity for other parasite species. PCR can detect intestinal parasites more accurately but is generally not feasible in resource-poor settings, at least not in peripheral labs. Hence, there is a need for a more field-friendly, sensitive approach for on-the-spot diagnosis of parasitic infections. PMID:28114314

A Review of Automatic Methods Based on Image Processing Techniques for Tuberculosis Detection from Microscopic Sputum Smear Images.

PubMed

Panicker, Rani Oomman; Soman, Biju; Saini, Gagan; Rajan, Jeny

2016-01-01

Tuberculosis (TB) is an infectious disease caused by the bacteria Mycobacterium tuberculosis. It primarily affects the lungs, but it can also affect other parts of the body. TB remains one of the leading causes of death in developing countries, and its recent resurgences in both developed and developing countries warrant global attention. The number of deaths due to TB is very high (as per the WHO report, 1.5 million died in 2013), although most are preventable if diagnosed early and treated. There are many tools for TB detection, but the most widely used one is sputum smear microscopy. It is done manually and is often time consuming; a laboratory technician is expected to spend at least 15 min per slide, limiting the number of slides that can be screened. Many countries, including India, have a dearth of properly trained technicians, and they often fail to detect TB cases due to the stress of a heavy workload. Automatic methods are generally considered as a solution to this problem. Attempts have been made to develop automatic approaches to identify TB bacteria from microscopic sputum smear images. In this paper, we provide a review of automatic methods based on image processing techniques published between 1998 and 2014. The review shows that the accuracy of algorithms for the automatic detection of TB increased significantly over the years and gladly acknowledges that commercial products based on published works also started appearing in the market. This review could be useful to researchers and practitioners working in the field of TB automation, providing a comprehensive and accessible overview of methods of this field of research.

Mathematical algorithm for the automatic recognition of intestinal parasites.

PubMed

Alva, Alicia; Cangalaya, Carla; Quiliano, Miguel; Krebs, Casey; Gilman, Robert H; Sheen, Patricia; Zimic, Mirko

2017-01-01

Parasitic infections are generally diagnosed by professionals trained to recognize the morphological characteristics of the eggs in microscopic images of fecal smears. However, this laboratory diagnosis requires medical specialists which are lacking in many of the areas where these infections are most prevalent. In response to this public health issue, we developed a software based on pattern recognition analysis from microscopi digital images of fecal smears, capable of automatically recognizing and diagnosing common human intestinal parasites. To this end, we selected 229, 124, 217, and 229 objects from microscopic images of fecal smears positive for Taenia sp., Trichuris trichiura, Diphyllobothrium latum, and Fasciola hepatica, respectively. Representative photographs were selected by a parasitologist. We then implemented our algorithm in the open source program SCILAB. The algorithm processes the image by first converting to gray-scale, then applies a fourteen step filtering process, and produces a skeletonized and tri-colored image. The features extracted fall into two general categories: geometric characteristics and brightness descriptions. Individual characteristics were quantified and evaluated with a logistic regression to model their ability to correctly identify each parasite separately. Subsequently, all algorithms were evaluated for false positive cross reactivity with the other parasites studied, excepting Taenia sp. which shares very few morphological characteristics with the others. The principal result showed that our algorithm reached sensitivities between 99.10%-100% and specificities between 98.13%- 98.38% to detect each parasite separately. We did not find any cross-positivity in the algorithms for the three parasites evaluated. In conclusion, the results demonstrated the capacity of our computer algorithm to automatically recognize and diagnose Taenia sp., Trichuris trichiura, Diphyllobothrium latum, and Fasciola hepatica with a high sensitivity and specificity.

GeneXpert MTB/RIF Testing in the Management of Patients with Active Tuberculosis; A Real Life Experience from Saudi Arabia.

PubMed

Omrani, Ali S; Al-Otaibi, Mohammed F; Al-Ateah, Souad M; Al-Onazi, Fahad M; Baig, Kamran; El-Khizzi, Noura A; Albarrak, Ali M

2014-03-01

GeneXpert MTB/RIF is a real-time PCR assay with established diagnostic performance in pulmonary and extra-pulmonary forms of tuberculosis. The aim of this study was to assess the contribution of GeneXpert MTB/RIF assay to the management of patients with any form of active tuberculosis in a single large tertiary center in Saudi Arabia, with a special focus on the impact on time to start of antituberculous therapy compared with Ziehl-Neelsen (ZN) smears and mycobacterial cultures. Clinical, radiological and laboratory records for all patients who were commenced on antituberculous therapy between March 2011 and February 2013 were retrospectively reviewed. A total of 140 patients were included, 38.6% of which had pulmonary tuberculosis. GeneXpert MTB/RIF was requested for only 39.2% of patients and was the only reason for starting antituberculous therapy for only 12.1%. The median time to a positive GeneXpert MTB/RIF result was 0 days (IQR 3) compared with 0 day (IQR 1) for smear microscopy (P > 0.999) and 22 days (IQR 21) for mycobacterial cultures (P < 0.001). No patients discontinued antituberculous therapy because of a negative GeneXpert MTB/RIF result. In a setting wherein physicians are highly experienced in the diagnosis and treatment of tuberculosis, GeneXpert MTB/RIF was remarkably under-utilized and had only a limited impact on decisions related to starting or stopping antituberculous therapy. Cost-effectiveness and clinical utility of routine testing of all smear-negative clinical samples submitted for tuberculosis investigations by GeneXpert MTB/RIF warrant further study.

The challenge of maintaining microscopist capacity at basic levels for malaria elimination in Jiangsu Province, China.

PubMed

Ding, Guisheng; Zhu, Guoding; Cao, Caiqun; Miao, Ping; Cao, Yuanyuan; Wang, Weiming; Gu, Yaping; Xu, Sui; Wang, Shengqiang; Zhou, Huayun; Cao, Jun

2018-04-12

Local malaria transmission has decreased rapidly since the National Malaria Elimination Action Plan was launched in China in 2010. However, imported malaria cases from Africa and Southeast Asia still occur in China due to overseas laborers. Diagnosis by microscopy is the gold standard for malaria and is used in most hospitals in China. However, the current capacity of microscopists to manage malaria cases in hospitals and public health facilities to meet the surveillance needs to eliminate and prevent the reintroduction of malaria is unknown. Malaria diagnoses were assessed by comparing the percentage of first visit and confirmed malaria diagnoses at Centers for Disease Control and Prevention (CDCs) and hospitals. The basic personnel information for public health departments and hospitals at different levels was investigated. The skills of microscopists for blood smear preparation and slide interpretation were also examined at the county and township levels. Inaccurate rate with 13.49% and 7.32%, respectively, in 2013 and 2014, from 341 and 355 reported cases from sub-provincial levels in Jiangsu province. Most of the 523 malaria cases reported in Nantong Prefecture from 2000 to 2014 involved patients who first visited county CDCs seeking treatment, however, none of these cases received confirmed diagnosis of malaria in townships or villages.The staff at county CDCs and hospitals with a higher education background performed better at making and interpreting blood smears than staff from townships. The network for malaria elimination in an entire province has been well established. However, an insufficient capacity for malaria diagnosis was observed, especially the preparing and reading the blood smears at the township and village levels, which is a challenge to achieving and maintaining malaria elimination.

Assessment of a gel-type chelating preparation containing 1-hydroxyethylidene-1, 1-bisphosphonate.

PubMed

Girard, S; Paqué, F; Badertscher, M; Sener, B; Zehnder, M

2005-11-01

To test an aqueous gel containing 1-hydroxyethylidene-1, 1-bisphosphonate (HEBP) regarding its interactions with sodium hypochlorite, its calcium binding capacity, and its potential in preventing the formation of a smear layer when used in conjunction with rotary root canal preparation. The experimental aqueous gel consisted of (w/v) 2% alginate, 3% aerosil, 10% Tween 80 and 18% HEBP. Interactions of gel components with hypochlorite were assessed using iodometric titration and monochromatic ultraviolet spectrometry. Two commercial paste-type chelators containing ethylenediaminetetraacetic acid (EDTA) and peroxide (RC-Prep and Glyde) served as controls. Calcium-binding capacities were measured in mixtures with a Ca2+ standard solution buffered at pH 10 using a calcium-selective measuring chain. Finally, root canals of 16 extracted single-rooted premolars per group were instrumented using ProFile instruments dipped in the experimental gel, RC-Prep, or nothing. Additionally, canals were rinsed with 10 mL of a 1% NaOCl solution during/after preparation. Smear scores in instrumented teeth were monitored using scanning electron microscopy. None of the experimental gel components showed short-term interactions with hypochlorite, whilst EDTA, peroxide, RC-Prep and Glyde immediately reduced the hypochlorite in solution. The experimental gel chelated 30 mg Ca2+ g-1, compared with 16 mg Ca2+ g-1 and 11 mg Ca2+ g-1 chelated by RC-Prep and Glyde respectively. Smear scores obtained with the experimental gel were significantly (P<0.05) lower than with RC-Prep in coronal and middle root thirds, whilst no differences were observed in apical root thirds. Under the conditions of this study, an HEBP gel appeared advantageous over currently available products.

Mathematical algorithm for the automatic recognition of intestinal parasites

PubMed Central

Alva, Alicia; Cangalaya, Carla; Quiliano, Miguel; Krebs, Casey; Gilman, Robert H.; Sheen, Patricia; Zimic, Mirko

2017-01-01

Parasitic infections are generally diagnosed by professionals trained to recognize the morphological characteristics of the eggs in microscopic images of fecal smears. However, this laboratory diagnosis requires medical specialists which are lacking in many of the areas where these infections are most prevalent. In response to this public health issue, we developed a software based on pattern recognition analysis from microscopi digital images of fecal smears, capable of automatically recognizing and diagnosing common human intestinal parasites. To this end, we selected 229, 124, 217, and 229 objects from microscopic images of fecal smears positive for Taenia sp., Trichuris trichiura, Diphyllobothrium latum, and Fasciola hepatica, respectively. Representative photographs were selected by a parasitologist. We then implemented our algorithm in the open source program SCILAB. The algorithm processes the image by first converting to gray-scale, then applies a fourteen step filtering process, and produces a skeletonized and tri-colored image. The features extracted fall into two general categories: geometric characteristics and brightness descriptions. Individual characteristics were quantified and evaluated with a logistic regression to model their ability to correctly identify each parasite separately. Subsequently, all algorithms were evaluated for false positive cross reactivity with the other parasites studied, excepting Taenia sp. which shares very few morphological characteristics with the others. The principal result showed that our algorithm reached sensitivities between 99.10%-100% and specificities between 98.13%- 98.38% to detect each parasite separately. We did not find any cross-positivity in the algorithms for the three parasites evaluated. In conclusion, the results demonstrated the capacity of our computer algorithm to automatically recognize and diagnose Taenia sp., Trichuris trichiura, Diphyllobothrium latum, and Fasciola hepatica with a high sensitivity and specificity. PMID:28410387

The Effect of Smear Layer Removal on Endodontic Outcomes

DTIC Science & Technology

2012-06-01

Sundqvist G. The antibacterial effect of sodium hypochlorite and EDTA in 60 cases of endodontic therapy. Int Endod J 1985;18:35-40. 11. Conner DA, Caplan...outcomes. KEY WORDS: Smear layer, smear layer removal, smear layer creation, EDTA, Sodium hypochlorite , Periapical index INTRODUCTION: The...identified the most efficient process for removal of the smear layer. Normal saline and sodium hypochlorite (NaOCl) have been shown not to remove

Role of 2-month sputum smears in predicting culture conversion in pulmonary tuberculosis.

PubMed

Su, W-J; Feng, J-Y; Chiu, Y-C; Huang, S-F; Lee, Y-C

2011-02-01

Sputum smears and culture conversion are frequently used to evaluate treatment response in pulmonary tuberculosis patients. Limited data are available on the evaluation of the correlation between under-treatment sputum smear results and culture conversion. This prospective study included sputum culture-proven pulmonary tuberculosis patients at six hospitals in Taiwan. At least two sets of sputum were collected at the completion of 8 weeks of TB treatment. The sensitivities and specificities of 2-month sputum smears were estimated based on culture conversion status. A total of 371 patients were enrolled for analysis. Factors associated with culture conversion included having a smear positive before treatment, presence of a cavity on radiography, rifampicin resistance and usage of the DOTS (directly observed therapy, short course) strategy. The sensitivities of 2-month sputum smears for culture conversion among all patients, initially smear-positive patients and initially smear-negative patients were 64.3, 71.4 and 38%, respectively, and the specificities were 81.6, 69.9 and 92.8%, respectively. In patients who were 2-month sputum smear-positive, the 2-month culture conversion rate was 80% if the patients were under DOTS and without cavitary lesions in radiograms. The predictive value of 2-month sputum smears in culture conversion was limited and highly influenced by clinical factors in pulmonary tuberculosis patients.

Knowledge of cervical cancer, attitude and husband’s support of Pap smear among multiparous women which have Pap’s smear examination in Aviati clinic Padang Bulan Medan

NASA Astrophysics Data System (ADS)

Feriyawati, L.; Anggraini, D. R.; Fitrie, A. A.; Anggreini, R. N.

2018-03-01

Cervical cancer is a serious health problem and stated as the second cause of death of woman worldwide. Several studies have noted a higher incidence of cervical cancer with increasing parity. Early detection with Pap smear is proven to reduce mortality of patients. Knowledge, attitude and husband’s support contributed to theled womanto follow Pap smear examination. This study explores the knowledge of cervical cancer, attitude and husband’ s support of Pap smearin multiparous women that have Pap smear examination. This research is a quantitative study with cross sectional approach recruited 50 respondents as multiparous women that have Pap smear examination inAviati Clinic, Padang Bulan, Medan. The data were collected by self-reports using structured objectives by questionnaires. The result of this study showed that 66% respondents have high knowledge of cervical cancer and 76% respondents have ahigh attitude of Pap smear, but they almost have low husband’s support of Pap smear including information support (62%), emotional support (46%) and real support (50%). This study has revealed that multiparous women that had Pap smear examination generally had high knowledge about cervical cancer and positive attitude about Pap smear, even most of them had low husband’s support.

Spot sputum samples are at least as good as early morning samples for identifying Mycobacterium tuberculosis.

PubMed

Murphy, Michael E; Phillips, Patrick P J; Mendel, Carl M; Bongard, Emily; Bateson, Anna L C; Hunt, Robert; Murthy, Saraswathi; Singh, Kasha P; Brown, Michael; Crook, Angela M; Nunn, Andrew J; Meredith, Sarah K; Lipman, Marc; McHugh, Timothy D; Gillespie, Stephen H

2017-10-27

The use of early morning sputum samples (EMS) to diagnose tuberculosis (TB) can result in treatment delay given the need for the patient to return to the clinic with the EMS, increasing the chance of patients being lost during their diagnostic workup. However, there is little evidence to support the superiority of EMS over spot sputum samples. In this new analysis of the REMoxTB study, we compare the diagnostic accuracy of EMS with spot samples for identifying Mycobacterium tuberculosis pre- and post-treatment. Patients who were smear positive at screening were enrolled into the study. Paired sputum samples (one EMS and one spot) were collected at each trial visit pre- and post-treatment. Microscopy and culture on solid LJ and liquid MGIT media were performed on all samples; those missing corresponding paired results were excluded from the analyses. Data from 1115 pre- and 2995 post-treatment paired samples from 1931 patients enrolled in the REMoxTB study were analysed. Patients were recruited from South Africa (47%), East Africa (21%), India (20%), Asia (11%), and North America (1%); 70% were male, median age 31 years (IQR 24-41), 139 (7%) co-infected with HIV with a median CD4 cell count of 399 cells/μL (IQR 318-535). Pre-treatment spot samples had a higher yield of positive Ziehl-Neelsen smears (98% vs. 97%, P = 0.02) and LJ cultures (87% vs. 82%, P = 0.006) than EMS, but there was no difference for positivity by MGIT (93% vs. 95%, P = 0.18). Contaminated and false-positive MGIT were found more often with EMS rather than spot samples. Surprisingly, pre-treatment EMS had a higher smear grading and shorter time-to-positivity, by 1 day, than spot samples in MGIT culture (4.5 vs. 5.5 days, P < 0.001). There were no differences in time to positivity in pre-treatment LJ culture, or in post-treatment MGIT or LJ cultures. Comparing EMS and spot samples in those with unfavourable outcomes, there were no differences in smear or culture results, and positive results were not detected earlier in Kaplan-Meier analyses in either EMS or spot samples. Our data do not support the hypothesis that EMS samples are superior to spot sputum samples in a clinical trial of patients with smear positive pulmonary TB. Observed small differences in mycobacterial burden are of uncertain significance and EMS samples do not detect post-treatment positives any sooner than spot samples.

Modelling the influence of noise of the image sensor for blood cells recognition in computer microscopy

NASA Astrophysics Data System (ADS)

Nikitaev, V. G.; Nagornov, O. V.; Pronichev, A. N.; Polyakov, E. V.; Dmitrieva, V. V.

2017-12-01

The first stage of diagnostics of blood cancer is the analysis of blood smears. The application of decision-making support systems would reduce the subjectivity of the diagnostic process and avoid errors, resulting in often irreversible changes in the patient's condition. In this regard, the solution of this problem requires the use of modern technology. One of the tools of the program classification of blood cells are texture features, and the task of finding informative among them is promising. The paper investigates the effect of noise of the image sensor to informative texture features with application of methods of mathematical modelling.

Low sensitivity of malaria rapid diagnostic tests stored at room temperature in the Brazilian Amazon Region.

PubMed

Gomes, Luciano T; Tada, Mauro S; Katsuragawa, Tony H; Povoa, Marinete M; Viana, Giselle Mr; Alecrim, Maria das Gracas C; De Santana-Filho, Frankllin S; Arcanjo, Ana Ruth L; Couto, Alvaro A R A; Calvosa, Vanja S P; Nery, Andreia F; Fontes, Cor J F

2013-03-14

In remote areas of the Amazon Region, diagnosis of malaria by microscopy is practically impossible. This study aimed to evaluate the performance of two rapid diagnostic tests (RDTs) targeting different malaria antigens stored at room temperature in the Brazilian Amazon Region. Performance of the OptiMal Pf/Pan test and ICT-Now Pf/Pan test was analyzed retrospectively in 1,627 and 1,602 blood samples, respectively. Tests were performed over a 15-month period. Kits were stored at room temperature in five community health centres located in the Brazilian Amazon Region. RDT results were compared with thick blood smear (TBS) results to determine sensitivity, specificity, and accuracy of the RDT. The sensitivities of the OptiMal Pf/Pan test were 79.7% for Plasmodium falciparum malaria diagnosis and 85.7% for non-P. falciparum infections. The results showed a crude agreement of 88.5% for P. falciparum, and 88.3% for non-P. falciparum infections (Kappa index = 0.74 and 0.75, respectively). For the ICT-Now Pf/Pan test (CI 95%), the sensitivities were 87.9% for P. falciparum malaria diagnosis and 72.5% for non-P. falciparum infection. Crude agreement between the ICT-Now Pf/Pan test and TBS was 91.4% for P. falciparum and 79.7% for non-P. falciparum infection. The Kappa index was 0.81 and 0.59 for the final diagnosis of P. falciparum and non-P. falciparum, respectively. Higher levels of parasitaemia were associated with higher crude agreement between RDT and TBS. The sensitivities of RDTs stored at room temperature over a 15-month period and performed in field conditions were lower than those previously reported.

Effectiveness of a computer-based tutorial for teaching how to make a blood smear.

PubMed

Preast, Vanessa; Danielson, Jared; Bender, Holly; Bousson, Maury

2007-09-01

Computer-aided instruction (CAI) was developed to teach veterinary students how to make blood smears. This instruction was intended to replace the traditional instructional method in order to promote efficient use of faculty resources while maintaining learning outcomes and student satisfaction. The purpose of this study was to evaluate the effect of a computer-aided blood smear tutorial on 1) instructor's teaching time, 2) students' ability to make blood smears, and 3) students' ability to recognize smear quality. Three laboratory sessions for senior veterinary students were taught using traditional methods (control group) and 4 sessions were taught using the CAI tutorial (experimental group). Students in the control group received a short demonstration and lecture by the instructor at the beginning of the laboratory and then practiced making blood smears. Students in the experimental group received their instruction through the self-paced, multimedia tutorial on a laptop computer and then practiced making blood smears. Data was collected from observation, interview, survey questionnaires, and smear evaluation by students and experts using a scoring rubric. Students using the CAI made better smears and were better able to recognize smear quality. The average time the instructor spent in the room was not significantly different between groups, but the quality of the instructor time was improved with the experimental instruction. The tutorial implementation effectively provided students and instructors with a teaching and learning experience superior to the traditional method of instruction. Using CAI is a viable method of teaching students to make blood smears.

Optimizing single-nanoparticle two-photon microscopy by in situ adaptive control of femtosecond pulses

DOE Office of Scientific and Technical Information (OSTI.GOV)

Li, Donghai; Deng, Yongkai; Chu, Saisai

2016-07-11

Single-nanoparticle two-photon microscopy shows great application potential in super-resolution cell imaging. Here, we report in situ adaptive optimization of single-nanoparticle two-photon luminescence signals by phase and polarization modulations of broadband laser pulses. For polarization-independent quantum dots, phase-only optimization was carried out to compensate the phase dispersion at the focus of the objective. Enhancement of the two-photon excitation fluorescence intensity under dispersion-compensated femtosecond pulses was achieved. For polarization-dependent single gold nanorod, in situ polarization optimization resulted in further enhancement of two-photon photoluminescence intensity than phase-only optimization. The application of in situ adaptive control of femtosecond pulse provides a way for object-orientedmore » optimization of single-nanoparticle two-photon microscopy for its future applications.« less

Correlative Stochastic Optical Reconstruction Microscopy and Electron Microscopy

PubMed Central

Kim, Doory; Deerinck, Thomas J.; Sigal, Yaron M.; Babcock, Hazen P.; Ellisman, Mark H.; Zhuang, Xiaowei

2015-01-01

Correlative fluorescence light microscopy and electron microscopy allows the imaging of spatial distributions of specific biomolecules in the context of cellular ultrastructure. Recent development of super-resolution fluorescence microscopy allows the location of molecules to be determined with nanometer-scale spatial resolution. However, correlative super-resolution fluorescence microscopy and electron microscopy (EM) still remains challenging because the optimal specimen preparation and imaging conditions for super-resolution fluorescence microscopy and EM are often not compatible. Here, we have developed several experiment protocols for correlative stochastic optical reconstruction microscopy (STORM) and EM methods, both for un-embedded samples by applying EM-specific sample preparations after STORM imaging and for embedded and sectioned samples by optimizing the fluorescence under EM fixation, staining and embedding conditions. We demonstrated these methods using a variety of cellular targets. PMID:25874453

Increased uptake of cervical screening by women with HIV infection in Auckland regardless of ethnicity, requirement for an interpreter or level of education.

PubMed

Lowe, Michele; Handy, Rupert; Ingram, Joan; Nisbet, Mitzi; Ritchie, Stephen; Thomas, Mark; Briggs, Simon

2016-07-15

Current guidelines recommend that women with HIV infection receive annual cervical smears. We evaluated the uptake of annual cervical smears by women with HIV infection under the care of the Infectious Disease Service at Auckland City Hospital. In an attempt to identify potential barriers to regularly receiving an annual cervical smear, we invited the women to complete a questionnaire. The responses from women who had regularly received an annual cervical smear were compared with those who had not. The proportion of women who had received a cervical smear increased from 44% in 2001, to 73% in 2010 (p=0.001). Ninety-three women (76%) completed the study questionnaire. No statistically significant differences were found in the questionnaire responses between the women who had regularly received an annual cervical smear and those who had not. The proportion of women in this cohort who received a cervical smear in 2010 is comparable with other studies of women with HIV infection in New Zealand and overseas. We have not been able to identify barriers that prevent women with HIV infection in Auckland regularly receiving an annual cervical smear. We plan to encourage women who have not received a cervical smear in the previous 2-year period to have a cervical smear performed when they attend the Infectious Disease Clinic, and will continue to notify the National Cervical Screening Programme that all women who are newly diagnosed with HIV infection should have an annual recall code attached to future cervical smear reports. We expect that these interventions will further increase the proportion of women with HIV infection in Auckland who receive an annual cervical smear.

The rate of sputum smear-positive tuberculosis after treatment default in a high-burden setting: a retrospective cohort study.

PubMed

Marx, Florian M; Dunbar, Rory; Enarson, Donald A; Beyers, Nulda

2012-01-01

High rates of recurrent tuberculosis after successful treatment have been reported from different high burden settings in Sub-Saharan Africa. However, little is known about the rate of smear-positive tuberculosis after treatment default. In particular, it is not known whether or not treatment defaulters continue to be or become again smear-positive and thus pose a potential for transmission of infection to others. To investigate, in a high tuberculosis burden setting, the rate of re-treatment for smear-positive tuberculosis among cases defaulting from standardized treatment compared to successfully treated cases. Retrospective cohort study among smear-positive tuberculosis cases treated between 1996 and 2008 in two urban communities in Cape Town, South Africa. Episodes of re-treatment for smear-positive tuberculosis were ascertained via probabilistic record linkage. Survival analysis and Poisson regression were used to compare the rate of smear-positive tuberculosis after treatment default to that after successful treatment. A total of 2,136 smear-positive tuberculosis cases were included in the study. After treatment default, the rate of re-treatment for smear-positive tuberculosis was 6.86 (95% confidence interval [CI]: 5.59-8.41) per 100 person-years compared to 2.09 (95% CI: 1.81-2.41) after cure (adjusted Hazard Ratio [aHR]: 3.97; 95% CI: 3.00-5.26). Among defaulters, the rate was inversely associated with treatment duration and sputum conversion prior to defaulting. Smear grade at start of the index treatment episode (Smear3+: aHR 1.61; 95%CI 1.11-2.33) was independently associated with smear-positive tuberculosis re-treatment, regardless of treatment outcome. In this high-burden setting, there is a high rate of subsequent smear-positive tuberculosis after treatment default. Treatment defaulters are therefore likely to contribute to the pool of infectious source cases in the community. Our findings underscore the importance of preventing treatment default, as a means of successful tuberculosis control in high-burden settings.

Optimal Design of Grid-Stiffened Panels and Shells With Variable Curvature

NASA Technical Reports Server (NTRS)

Ambur, Damodar R.; Jaunky, Navin

2001-01-01

A design strategy for optimal design of composite grid-stiffened structures with variable curvature subjected to global and local buckling constraints is developed using a discrete optimizer. An improved smeared stiffener theory is used for the global buckling analysis. Local buckling of skin segments is assessed using a Rayleigh-Ritz method that accounts for material anisotropy and transverse shear flexibility. The local buckling of stiffener segments is also assessed. Design variables are the axial and transverse stiffener spacing, stiffener height and thickness, skin laminate, and stiffening configuration. Stiffening configuration is herein defined as a design variable that indicates the combination of axial, transverse and diagonal stiffeners in the stiffened panel. The design optimization process is adapted to identify the lightest-weight stiffening configuration and stiffener spacing for grid-stiffened composite panels given the overall panel dimensions. in-plane design loads, material properties. and boundary conditions of the grid-stiffened panel or shell.

Cost-effectiveness of microscopy of urethral smears for asymptomatic Mycoplasma genitalium urethritis in men in England.

PubMed

Sutton, Andrew J; Roberts, Tracy E; Jackson, Louise; Saunders, John; White, Peter J; Birger, Ruthie; Estcourt, Claudia

2018-01-01

The objective was to determine whether or not the limited use of urethral microscopy to diagnose asymptomatic and symptomatic non-chlamydial, non-gonococcal urethritis (NCNGU) in men is a cost-effective strategy to avert pelvic inflammatory disease (PID), ectopic pregnancy or infertility in female partners. Outputs from a transmission dynamic model of NCNGU in a population of 16-30 year olds in England simulating the number of consultations, PID cases and patients treated over time amongst others, were used along with secondary data to undertake a cost-effectiveness analysis carried out from a health care provider perspective. The main outcome measure was cost per case of PID averted. A secondary outcome measure was cost per major outcome averted, where a major outcome is a case of symptomatic PID, ectopic pregnancy, or infertility. Offering a limited number of asymptomatic men urethral microscopy was more effective than the current practice of no microscopy in terms of reducing the number of cases of PID with an incremental cost-effectiveness ratio of £15,700, meaning that an investment of £15,800 is required to avert one case of PID. For major outcomes averted, offering some asymptomatic men urethral microscopy was again found to be more effective than no microscopy, but here an investment of £49,900 is required to avert one major outcome. Testing asymptomatic men for NCNGU in a small number of genitourinary medicine settings in England is not cost-effective, and thus by maintaining the current practice of not offering this patient group microscopy, this continues to make savings for the health care provider.

Assessing the reliability of microscopy and rapid diagnostic tests in malaria diagnosis in areas with varying parasite density among older children and adult patients in Nigeria.

PubMed

Ayogu, E E; Ukwe, C V; Nna, E O

2016-01-01

Current malaria control strategies are based on early diagnosis and appropriate treatment of malaria cases. The study aimed at comparing the performance of blood film microscopy and rapid diagnostic test (RDT) in Plasmodium falciparum detection in patients ≥6 years of age. A total of 154 consecutive pyretic patients aged 6-62 years were enrolled, sampled, and tested for malaria using RDT (first response) and microscopy by Giemsa staining. Genomic DNA was extracted after saponin hemolysis and nested polymerase chain reaction (PCR) was used to detect Plasmodium falciparum. The endpoints were sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). Of the 154 patients, 80 (51.9%) had fever of ≥37.5°C. 106 (68.8%) were positive by First response® , 132 (85.7%) by microscopy, and 121 (78.6%) by PCR. The sensitivity, specificity, PPV, and NPV of first response compared to microscopic method were 82.2%, 100.0%, 100.0%, and 34.3%, respectively, while it was 75.4%, 75.0%, 95.3%, and 31.2%, respectively, when compared to PCR. The sensitivity, specificity, PPV, and NPV of the microscopic method compared to PCR were 92.3%, 50.0%, 90.91%, and 54.5%, respectively. There was a significant difference in the performance of RDT and film microscopy methods (P ≤ 0.05). Microscopy performed better and is more reliable than first response (RDT) in areas with low parasite density among patients ≥6 years of age. Rapid diagnostic tests could be useful in aareas with high parasite density as an alternative to smear microscopy.

[Tracing investigation and diagnosis of one transfusion-transmitted malaria case in Zhejiang Province].

PubMed

Ruan, Wei; Lei, Yong-Liang; Yao, Li-Nong; Zhang, Ling-Ling; Liu, Xiao-Hong; Xiang, Xiao-Qing; Mei, Jian-Hua; Zhu, Hai-Bo; Yu, Yang; Zeng, Chang-You

2014-02-01

To identify the sources of infection and the mode of transmission of a malaria case with unknown origin. Clinical data of the case were collected and the epidemiological investigation was conducted. The blood samples of the patient and the suspected infection source (blood donor) were detected by microscopy, rapid diagnostic test strip (RDT) and nested PCR. The patient did not visited malaria endemic areas. After a blood transfusion, the patient had chills and fever, and was confirmed as falciparum malaria by microscopy with bone marrow and peripheral blood smears and RDT. The blood donor was a worker returned from Africa. Before blood donation she was sick like malaria carrier, and took anti-malarial drug. She was then confirmed as falciparum malaria by RDT and microscopy. The blood samples from the patient and the blood donor were diagnosed as falciparum malaria by nested PCR, and the similarity of the small subunit rRNA (SSU rRNA) sequence was 100%, showing they were mix-infected with K1 and MAD20 genotypes of Plasmodium falciparum. This patient is confirmed P. falciparum infection via blood transfusion from a donor who returned from Africa.

Prevalence of Malaria in Pregnant Women in Lagos, South-West Nigeria

PubMed Central

Agomo, Chimere O.; Anorlu, Rose I.; Agomo, Philip U.

2009-01-01

Prevalence rates reported for malaria in pregnancy in Nigeria vary considerably. The accuracy of results of malaria diagnosis is dependent on training, experience, and motivation of the microscopist as well as the laboratory facility available. Results of training programmes on malaria microscopy have shown low levels of sensitivity and specificity of those involved in malaria diagnosis routinely and for research. This study was done to ascertain the true prevalence of malaria in pregnancy in Lagos, South-West Nigeria. A total of 1,084 pregnant women were recruited into this study. Blood smears stained with Giemsa were used for malaria diagnosis by light microscopy. Malaria infection during pregnancy presents mostly as asymptomatic infection. The prevalence of malaria in this population was 7.7% (95% confidence interval; 6.2-9.4%). Factors identified to increase the risk of malaria infection include young maternal age (< 20 years), and gravidity (primigravida). In conclusion, this study exposes the over-diagnosis of malaria in pregnancy and the need for training and retraining of laboratory staffs as well as establishing the malaria diagnosis quality assurance programme to ensure the accuracy of malaria microscopy results at all levels. PMID:19488427

Detection and quantification of Plasmodium falciparum in blood samples using quantitative nucleic acid sequence-based amplification.

PubMed

Schoone, G J; Oskam, L; Kroon, N C; Schallig, H D; Omar, S A

2000-11-01

A quantitative nucleic acid sequence-based amplification (QT-NASBA) assay for the detection of Plasmodium parasites has been developed. Primers and probes were selected on the basis of the sequence of the small-subunit rRNA gene. Quantification was achieved by coamplification of the RNA in the sample with one modified in vitro RNA as a competitor in a single-tube NASBA reaction. Parasite densities ranging from 10 to 10(8) Plasmodium falciparum parasites per ml could be demonstrated and quantified in whole blood. This is approximately 1,000 times more sensitive than conventional microscopy analysis of thick blood smears. Comparison of the parasite densities obtained by microscopy and QT-NASBA with 120 blood samples from Kenyan patients with clinical malaria revealed that for 112 of 120 (93%) of the samples results were within a 1-log difference. QT-NASBA may be especially useful for the detection of low parasite levels in patients with early-stage malaria and for the monitoring of the efficacy of drug treatment.

Lensfree on-chip microscopy over a wide field-of-view using pixel super-resolution

PubMed Central

Bishara, Waheb; Su, Ting-Wei; Coskun, Ahmet F.; Ozcan, Aydogan

2010-01-01

We demonstrate lensfree holographic microscopy on a chip to achieve ~0.6 µm spatial resolution corresponding to a numerical aperture of ~0.5 over a large field-of-view of ~24 mm2. By using partially coherent illumination from a large aperture (~50 µm), we acquire lower resolution lensfree in-line holograms of the objects with unit fringe magnification. For each lensfree hologram, the pixel size at the sensor chip limits the spatial resolution of the reconstructed image. To circumvent this limitation, we implement a sub-pixel shifting based super-resolution algorithm to effectively recover much higher resolution digital holograms of the objects, permitting sub-micron spatial resolution to be achieved across the entire sensor chip active area, which is also equivalent to the imaging field-of-view (24 mm2) due to unit magnification. We demonstrate the success of this pixel super-resolution approach by imaging patterned transparent substrates, blood smear samples, as well as Caenoharbditis Elegans. PMID:20588977

Microwave-accelerated cytochemical stains for the image analysis and the electron microscopic examination of light microscopy diagnostic slides.

PubMed

Hanker, J; Giammara, B

1993-01-01

Recent studies in our laboratories have shown how microwave (MW) irradiation can accelerate a number of tissue-processing techniques, especially staining, to aid in the preparation of single specimens on glass microscope slides or coverslips for examination by light microscopy (and electron microscopy, if required) for diagnostic purposes. Techniques have been developed, which give permanently stained preparations, that can be studied initially by light microscopy, their areas of interest mapped, and computer-automated image analysis performed to obtain quantitative information. This is readily performed after MW-accelerated staining with silver methenamine by the Giammara-Hanker PATS or PATS-TS reaction. This variation of the PAS reaction gives excellent markers for specific infectious agents such as lipopolysaccharides for gram-negative bacteria or mannans for fungi. It is also an excellent stain for glycogen and basement membranes and an excellent marker for type III collagen or reticulin in the endoneurium or perineurium of peripheral nerve or in the capillary walls. Our improved MW-accelerated Feulgen reaction with silver methenamine for nuclear DNA is useful to show the nuclei of bacteria and fungi as well as of cells they are infecting. Improved coating and penetration of tissue surfaces by thiocarbohydrazide bridging of ruthenium red, applied under MW-acceleration, render biologic specimens sufficiently conductive for SEM so that sputter coating with gold is unnecessary. The specimens treated with these highly visible electron-opaque stains can be screened with the light microscope after mounting in polyethylene glycol (PEG) and the structures or areas selected for EM study are mapped with a Micro-Locator slide. After removal of the water soluble PEG the specimens are remounted in the usual EM media for scanning electron microscopy (SEM) or transmission electron microscopy (TEM) study of the mapped areas. By comparing duplicate smears from areas of infection, such as two coverslips of buffy coat smears of blood from a patient with septicemia, the microorganisms responsible can occasionally be classified for antimicrobial therapy long before culture results are available; gram-negative bacteria are positive with the Giammara-Hanker PATS-TS stain, and gram-positive bacteria are positive with the SIGMA HT40 Gram stain. The gram-positive as well as gram-negative bacteria are both initially stained by the crystal violet component of the Gram stain. The crystal violet stain is readily removed from the gram-negative (but not the gram-positive) bacteria when the specimens are rinsed with alcohol/acetone. If this rinse step is omitted, the crystal violet remains attached to both gram-negative and gram-positive bacteria. It can then be rendered insoluble, electron-opaque, and conductive by treatment with silver methenamine solution under MW-irradiation. This metallized crystal violet is a more effective silver stain than the PATS-TS stain for a number of gram-negative spirochetes such as Treponema pallidum, the microbe that causes syphilis.

Locally-smeared operator product expansions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Monahan, Christopher; Orginos, Kostantinos

2014-12-01

We propose a "locally-smeared Operator Product Expansion" (sOPE) to decompose non-local operators in terms of a basis of locally-smeared operators. The sOPE formally connects nonperturbative matrix elements of smeared degrees of freedom, determined numerically using the gradient flow, to non-local operators in the continuum. The nonperturbative matrix elements do not suffer from power-divergent mixing on the lattice, provided the smearing scale is kept fixed in the continuum limit. The presence of this smearing scale prevents a simple connection to the standard operator product expansion and therefore requires the construction of a two-scale formalism. We demonstrate the feasibility of our approachmore » using the example of real scalar field theory.« less

On the ground state of Yang-Mills theory

NASA Astrophysics Data System (ADS)

Bakry, Ahmed S.; Leinweber, Derek B.; Williams, Anthony G.

2011-08-01

We investigate the overlap of the ground state meson potential with sets of mesonic-trial wave functions corresponding to different gluonic distributions. We probe the transverse structure of the flux tube through the creation of non-uniform smearing profiles for the string of glue connecting two color sources in Wilson loop operator. The non-uniformly UV-regulated flux-tube operators are found to optimize the overlap with the ground state and display interesting features in the ground state overlap.

Cost-Effectiveness of Automated Digital Microscopy for Diagnosis of Active Tuberculosis.

PubMed

Jha, Swati; Ismail, Nazir; Clark, David; Lewis, James J; Omar, Shaheed; Dreyer, Andries; Chihota, Violet; Churchyard, Gavin; Dowdy, David W

2016-01-01

Automated digital microscopy has the potential to improve the diagnosis of tuberculosis (TB), particularly in settings where molecular testing is too expensive to perform routinely. The cost-effectiveness of TB diagnostic algorithms using automated digital microscopy remains uncertain. Using data from a demonstration study of an automated digital microscopy system (TBDx, Applied Visual Systems, Inc.), we performed an economic evaluation of TB diagnosis in South Africa from the health system perspective. The primary outcome was the incremental cost per new TB diagnosis made. We considered costs and effectiveness of different algorithms for automated digital microscopy, including as a stand-alone test and with confirmation of positive results with Xpert MTB/RIF ('Xpert', Cepheid, Inc.). Results were compared against both manual microscopy and universal Xpert testing. In settings willing to pay $2000 per incremental TB diagnosis, universal Xpert was the preferred strategy. However, where resources were not sufficient to support universal Xpert, and a testing volume of at least 30 specimens per day could be ensured, automated digital microscopy with Xpert confirmation of low-positive results could facilitate the diagnosis of 79-84% of all Xpert-positive TB cases, at 50-60% of the total cost. The cost-effectiveness of this strategy was $1280 per incremental TB diagnosis (95% uncertainty range, UR: $340-$3440) in the base case, but improved under conditions likely reflective of many settings in sub-Saharan Africa: $677 per diagnosis (95% UR: $450-$935) when sensitivity of manual smear microscopy was lowered to 0.5, and $956 per diagnosis (95% UR: $40-$2910) when the prevalence of multidrug-resistant TB was lowered to 1%. Although universal Xpert testing is the preferred algorithm for TB diagnosis when resources are sufficient, automated digital microscopy can identify the majority of cases and halve the cost of diagnosis and treatment when resources are more scarce and multidrug-resistant TB is not common.

Is PCR the Next Reference Standard for the Diagnosis of Schistosoma in Stool? A Comparison with Microscopy in Senegal and Kenya.

PubMed

Meurs, Lynn; Brienen, Eric; Mbow, Moustapha; Ochola, Elizabeth A; Mboup, Souleymane; Karanja, Diana M S; Secor, W Evan; Polman, Katja; van Lieshout, Lisette

2015-01-01

The current reference test for the detection of S. mansoni in endemic areas is stool microscopy based on one or more Kato-Katz stool smears. However, stool microscopy has several shortcomings that greatly affect the efficacy of current schistosomiasis control programs. A highly specific multiplex real-time polymerase chain reaction (PCR) targeting the Schistosoma internal transcriber-spacer-2 sequence (ITS2) was developed by our group a few years ago, but so far this PCR has been applied mostly on urine samples. Here, we performed more in-depth evaluation of the ITS2 PCR as an alternative method to standard microscopy for the detection and quantification of Schistosoma spp. in stool samples. Microscopy and PCR were performed in a Senegalese community (n = 197) in an area with high S. mansoni transmission and co-occurrence of S. haematobium, and in Kenyan schoolchildren (n = 760) from an area with comparatively low S. mansoni transmission. Despite the differences in Schistosoma endemicity the PCR performed very similarly in both areas; 13-15% more infections were detected by PCR when comparing to microscopy of a single stool sample. Even when 2-3 stool samples were used for microscopy, PCR on one stool sample detected more infections, especially in people with light-intensity infections and in children from low-risk schools. The low prevalence of soil-transmitted helminthiasis in both populations was confirmed by an additional multiplex PCR. The ITS2-based PCR was more sensitive than standard microscopy in detecting Schistosoma spp. This would be particularly useful for S. mansoni detection in low transmission areas, and post-control settings, and as such improve schistosomiasis control programs, epidemiological research, and quality control of microscopy. Moreover, it can be complemented with other (multiplex real-time) PCRs to detect a wider range of helminths and thus enhance effectiveness of current integrated control and elimination strategies for neglected tropical diseases.

Field Comparison of the Sampling Efficacy of Two Smear Media: Cotton Fiber and Kraft Paper

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hogue, M.G.

Two materials were compared in field tests at the Defense Waste Processing Facility: kraft paper (a strong, brown paper made from wood pulp prepared with a sodium sulfate solution) and cotton fiber. Based on a sampling of forty-six pairs of smears, the cotton fiber smears provide a greater sensitivity. The cotton fiber smears collected an average of forty-four percent more beta activity than the kraft paper smears and twenty-nine percent more alpha activity. Results show a greater sensitivity with cotton fiber over kraft paper at the 95 percent confidence level. Regulatory requirements for smear materials are vague. The data demonstratemore » that the difference in sensitivity of smear materials could lead to a large difference in reported results that are subsequently used for meeting shipping regulations or evaluating workplace contamination levels.« less

Rates of abnormalities and infectious agents in cervical smears from female inmates in Texas: comparison with private and university clinic patients.

PubMed

Logrono, R; Wong, J Y

1999-09-01

Inmates are generally considered a high-risk population for gynecologic neoplasia and sexually transmitted diseases. Cervical smears from prisoners of the Texas Department of Corrections (TDC) were expected initially to have higher rates of cellular abnormalities and infectious agents than do smears from the general population. The cytologic findings from 25,522 TDC gynecologic smears were compared with those of 6883 cases from The University of Texas Medical Branch (UTMB) affiliated physician private clinics, and with 56,178 from the UTMB hospital clinics. The period of study was from September 1995 to February 1998. This study revealed a 5.23% higher rate of abnormalities for TDC gynecologic smears as compared with that for the private clinic smears. However, the TDC rate of abnormalities was unexpectedly 1.08% lower than that for the UTMB clinic smears. These unexpected findings were probably the result of a more selected high-risk population referred to the UTMB clinics. The TDC smears showed also the highest incidence of trichomoniasis.

Atomic force microscopy and scanning electron microscopy evaluation of efficacy of scaling and root planing using magnification: A randomized controlled clinical study

PubMed Central

Mohan, Ranjana; Agrawal, Sudhanshu; Gundappa, Mohan

2013-01-01

Aim: A randomized controlled clinical study was undertaken to evaluate the effectiveness of scaling and root planing (SRP) by using Magnifying Loupes (ML) and dental operating microscope (DOM). Materials and Methods: A total of 90 human teeth scheduled for extraction from 18 patients aged between 25 and 65 years suffering from generalized chronic severe periodontitis were randomly assigned to three treatment groups. Group 1 consisted SRP performed without using magnification (unaided), Group 2-SRP with ML and Group 3-SRP with DOM. Following extractions, samples were prepared for (i) evaluation of surface topography by atomic force microscopy, (ii) presence of smear layer, debris by scanning electron microscopy (iii) elemental analysis by energy dispersive X-ray analysis. Data was subjected to statistical analysis using analysis of variance, post-hoc (Tukey-HSD) and Chi-square test. Results: Statistically significant (P < 0.001) difference was found among the different treatment groups. Group 3 was the best while Group 1 was the least effective technique for SRP. Order of efficacy in terms of the surface was found to be - Palatal < Lingual < Distal ≃ Mesial < Buccal. Efficiency in mandibular to maxillary teeth was found to be significant (P < 0.05), also anterior to posterior teeth (P < 0.05). Conclusion: Magnification tools significantly enhance the efficacy of supragingival and subgingival SRP. PMID:24124292

Comparison of specimen adequacy and smear quality in oral smears prepared by manual liquid-based cytology and conventional methods

PubMed Central

Shukla, Surabhi; Einstein, A; Shukla, Abhilasha; Mishra, Deepika

2015-01-01

Background: Liquid-based cytology (LBC), recommended in the mass screening of potentially malignant cervical and oral lesions, suffers from high cost owing to the use of expensive automated devices and materials. Considering the need for cost-effective LBC techniques, we evaluated the efficacy of an inexpensive manual LBC (MLBC) technique against conventional cytological technique in terms of specimen adequacy and smear quality of oral smears. Materials and Methods: Cytological samples were collected from 21 patients using a cytobrush device. After preparation of a conventional smear, the brush containing the remaining sample was immersed in the preservative vial. The preserved material was processed by an MLBC technique and subsequently, direct smears were made from the prepared cell button. Both conventional and MLBC smears were stained by routine Papanicolaou technique and evaluated by an independent observer for the thickness of the smear, cellular distribution, resolution/clarity of cells, cellular staining characteristics and the presence of unsatisfactory background/artifacts. Each parameter was graded as satisfactory; or satisfactory, but limited; or unsatisfactory. Chi-square test was used to compare the values obtained (significance set at P ≤ 0.05). Results: MLBC technique produced a significant number of satisfactory smears with regard to cell distribution, clarity/resolution, staining characteristics and background/artifacts compared to conventional methods. Conclusions: MLBC is a cost-effective cytological technique that may produce oral smears with excellent cytomorphology and longer storage life. PMID:26980958

Smear layer-deproteinizing improves bonding of one-step self-etch adhesives to dentin.

PubMed

Thanatvarakorn, Ornnicha; Prasansuttiporn, Taweesak; Thittaweerat, Suppason; Foxton, Richard M; Ichinose, Shizuko; Tagami, Junji; Hosaka, Keiichi; Nakajima, Masatoshi

2018-03-01

Smear layer deproteinizing was proved to reduce the organic phase of smear layer covered on dentin surface. It was shown to eliminate hybridized smear layer and nanoleakage expression in resin-dentin bonding interface of two-step self-etch adhesive. This study aimed to investigate those effects on various one-step self-etch adhesives. Four different one-step self-etch adhesives were used in this study; SE One (SE), Scotchbond™ Universal (SU), BeautiBond Multi (BB), and Bond Force (BF). Flat human dentin surfaces with standardized smear layer were prepared. Smear layer deproteinizing was carried out by the application of 50ppm hypochlorous acid (HOCl) on dentin surface for 15s followed by Accel ® (p-toluenesulfinic acid salt) for 5s prior to adhesive application. No surface pretreatment was used as control. Microtensile bond strength (μTBS) and nanoleakage under TEM observation were investigated. The data were analyzed by two-way ANOVA and Tukey's post-hoc test and t-test at the significant level of 0.05. Smear layer deproteinizing significantly improved μTBS of SE, SU, and BB (p<0.001). Hybridized smear layer observed in control groups of SE, BB, and BF, and reticular nanoleakage presented throughout the hybridized complex in control groups of BB and BF were eliminated upon the smear layer deproteinizing. Smear layer deproteinizing by HOCl and Accel ® application could enhance the quality of dentin for bonding to one-step self-etch adhesives, resulting in the improving μTBS, eliminating hybridized smear layer and preventing reticular nanoleakage formation in resin-dentin bonding interface. Copyright © 2018 The Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Human bloodstains on bone artefacts: an SEM intra- and inter-sample comparative study using ratite bird tibiotarsus.

PubMed

Hortolà, Policarp

2016-11-01

Apart from their forensic significance in crime investigation, human bloodstains have an anthropological interest due to their occurrence on certain traditional weapons and ritual objects. Previously, a guiding study of erythrocytes in experimental samples including domestic sheep (Ovis aries) tibia was carried out using a scanning electron microscope (SEM). Here, a comparative SEM study to reveal the potential differences in bloodstain surface morphology as a function of intra-sample (smear region) and inter-sample (individual smear, smearing mechanism, bone origin) parameters is reported. A fragment of emu (Dromaius novaehollandiae) tibiotarsus was smeared with an adult man's peripheral blood. After air-drying and storing indoors, the boundary and neighbouring inner areas of the three individual bloodstains obtained were examined via secondary electrons in a variable-pressure SEM working in low-vacuum mode. As a whole, desiccation microcracks were present, the limits between the smear and the substrate appeared poorly defined, and no erythrocyte negative replicas were observed in the examined areas. In addition, a putative fibrin network, more or less embedded in the dried plasma matrix, was observed in the smears' boundary. Regarding the smear region in sliding smears, the periphery and boundary revealed to be different, while the head and tail were similar. Considering individual sliding smears, they had similar characteristics. Relating to the smear region as a function of the smearing mechanism, the periphery was different whether sliding or touching, while the boundary was similar in sliding and touching smears. Concerning the smear region as a function of the bone origin, the periphery revealed to be similar in both ratite and mammalian bone, while the boundary did different in ratite and mammalian bone. The results of this study show that SEM examination can be used fruitfully to detect bloodstains on ratite bone. Combined with previous SEM results in domestic sheep bone, they suggest, further, that blood remains can be detected on objects made of bone irrespectively of the mammalian or ratite origin of this raw material. Copyright © 2016 Elsevier Ltd. All rights reserved.

Incremental Yield of Including Determine-TB LAM Assay in Diagnostic Algorithms for Hospitalized and Ambulatory HIV-Positive Patients in Kenya.

PubMed

Huerga, Helena; Ferlazzo, Gabriella; Bevilacqua, Paolo; Kirubi, Beatrice; Ardizzoni, Elisa; Wanjala, Stephen; Sitienei, Joseph; Bonnet, Maryline

2017-01-01

Determine-TB LAM assay is a urine point-of-care test useful for TB diagnosis in HIV-positive patients. We assessed the incremental diagnostic yield of adding LAM to algorithms based on clinical signs, sputum smear-microscopy, chest X-ray and Xpert MTB/RIF in HIV-positive patients with symptoms of pulmonary TB (PTB). Prospective observational cohort of ambulatory (either severely ill or CD4<200cells/μl or with Body Mass Index<17Kg/m2) and hospitalized symptomatic HIV-positive adults in Kenya. Incremental diagnostic yield of adding LAM was the difference in the proportion of confirmed TB patients (positive Xpert or MTB culture) diagnosed by the algorithm with LAM compared to the algorithm without LAM. The multivariable mortality model was adjusted for age, sex, clinical severity, BMI, CD4, ART initiation, LAM result and TB confirmation. Among 474 patients included, 44.1% were severely ill, 69.6% had CD4<200cells/μl, 59.9% had initiated ART, 23.2% could not produce sputum. LAM, smear-microscopy, Xpert and culture in sputum were positive in 39.0% (185/474), 21.6% (76/352), 29.1% (102/350) and 39.7% (92/232) of the patients tested, respectively. Of 156 patients with confirmed TB, 65.4% were LAM positive. Of those classified as non-TB, 84.0% were LAM negative. Adding LAM increased the diagnostic yield of the algorithms by 36.6%, from 47.4% (95%CI:39.4-55.6) to 84.0% (95%CI:77.3-89.4%), when using clinical signs and X-ray; by 19.9%, from 62.2% (95%CI:54.1-69.8) to 82.1% (95%CI:75.1-87.7), when using clinical signs and microscopy; and by 13.4%, from 74.4% (95%CI:66.8-81.0) to 87.8% (95%CI:81.6-92.5), when using clinical signs and Xpert. LAM positive patients had an increased risk of 2-months mortality (aOR:2.7; 95%CI:1.5-4.9). LAM should be included in TB diagnostic algorithms in parallel to microscopy or Xpert request for HIV-positive patients either ambulatory (severely ill or CD4<200cells/μl) or hospitalized. LAM allows same day treatment initiation in patients at higher risk of death and in those not able to produce sputum.

Preventing Infectious Pulmonary Tuberculosis Among Foreign-Born Residents of the United States

PubMed Central

Katz, Dolly; Ghosh, Smita; Blumberg, Henry; Tamhane, Ashutosh; Sevilla, Anna; Reves, Randall

2015-01-01

Objectives. We described risk factors associated with infectious tuberculosis (TB) and missed TB-prevention opportunities in foreign-born US residents, who account for almost two thirds of the nation’s TB patients. Methods. In a cross-sectional study at 20 US sites of foreign-born persons diagnosed with TB in 2005 through 2006, we collected results of sputum smear microscopy for acid-fast bacilli (a marker for infectiousness) and data on visa status, sociodemographics, TB-related care seeking, and latent TB infection (LTBI) diagnosis opportunities. Results. Among 980 persons with pulmonary TB who reported their visa status, 601 (61%) were legal permanent residents, 131 (13.4%) had temporary visas, and 248 (25.3%) were undocumented. Undocumented persons were more likely than permanent residents to have acid-fast bacilli–positive smears at diagnosis (risk ratio = 1.3; 95% confidence interval = 1.2, 1.4). Of those diagnosed 1 year or more after arrival, 57.3% reported LTBI screening opportunities; fewer than 25% actually were. Undocumented persons reported fewer LTBI screening opportunities and were less likely to be tested. Conclusions. Progress toward TB elimination in the United States depends upon expanding opportunities for regular medical care and promotion of LTBI screening and treatment among foreign-born persons. PMID:26180947

[Microbiological diagnosis of imported malaria].

PubMed

Torrús, Diego; Carranza, Cristina; Manuel Ramos, José; Carlos Rodríguez, Juan; Rubio, José Miguel; Subirats, Mercedes; Ta-Tang, Thuy-Huong

2015-07-01

Current diagnosis of malaria is based on the combined and sequential use of rapid antigen detection tests (RDT) of Plasmodium and subsequent visualization of the parasite stained with Giemsa solution in a thin and thick blood smears. If an expert microscopist is not available, should always be a sensitive RDT to rule out infection by Plasmodium falciparum, output the result immediately and prepare thick smears (air dried) and thin extensions (fixed with methanol) for subsequent staining and review by an expert microscopist. The RDT should be used as an initial screening test, but should not replace microscopy techniques, which should be done in parallel. The diagnosis of malaria should be performed immediately after clinical suspicion. The delay in laboratory diagnosis (greater than 3 hours) should not prevent the initiation of empirical antimalarial treatment if the probability of malaria is high. If the first microscopic examination and RDT are negative, they must be repeated daily in patients with high suspicion. If suspicion remains after three negative results must be sought the opinion of an tropical diseases expert. Genomic amplification methods (PCR) are useful as confirmation of microscopic diagnosis, to characterize mixed infections undetectable by other methods, and to diagnose asymptomatic infections with submicroscopic parasitaemia. Copyright © 2015 Elsevier España, S.L.U. All rights reserved.

Automated plasmodia recognition in microscopic images for diagnosis of malaria using convolutional neural networks

NASA Astrophysics Data System (ADS)

Krappe, Sebastian; Benz, Michaela; Gryanik, Alexander; Tannich, Egbert; Wegner, Christine; Stamminger, Marc; Wittenberg, Thomas; Münzenmayer, Chrisitan

2017-03-01

Malaria is one of the world's most common and serious tropical diseases, caused by parasites of the genus plasmodia that are transmitted by Anopheles mosquitoes. Various parts of Asia and Latin America are affected but highest malaria incidence is found in Sub-Saharan Africa. Standard diagnosis of malaria comprises microscopic detection of parasites in stained thick and thin blood films. As the process of slide reading under the microscope is an error-prone and tedious issue we are developing computer-assisted microscopy systems to support detection and diagnosis of malaria. In this paper we focus on a deep learning (DL) approach for the detection of plasmodia and the evaluation of the proposed approach in comparison with two reference approaches. The proposed classification schemes have been evaluated with more than 180,000 automatically detected and manually classified plasmodia candidate objects from so-called thick smears. Automated solutions for the morphological analysis of malaria blood films could apply such a classifier to detect plasmodia in the highly complex image data of thick smears and thereby shortening the examination time. With such a system diagnosis of malaria infections should become a less tedious, more reliable and reproducible and thus a more objective process. Better quality assurance, improved documentation and global data availability are additional benefits.

Fatal spirochetosis due to a relapsing fever-like Borrelia sp. in northern spotted owl

USGS Publications Warehouse

Thomas, N.J.; Bunikis, J.; Barbour, A.G.; Wolcott, M.J.

2002-01-01

Acute septicemic spirochetosis was diagnosed in an adult male northern spotted owl (Strix occidentalis caurina) found dead in Kittitas County, Washington, USA. Gross necropsy findings included marked enlargement of the liver and spleen and serofibrinous deposits on the serous membranes lining the body cavities and the pericardial and perihepatic sacs. Microscopic observations included macrophage infiltration in the liver and spleen with mild thrombosis and multifocal necrosis, as well as hemorrhage and acute inflammation in the choroid plexus of the brain. No viruses or pathogenic bacteria were isolated from brain, liver, or spleen, and no parasites were found in blood smears or impression smears of the liver. Chlamydial culture attempts were unsuccessful and no chlamydial antibodies were detected in serum. In silver-stained microscopic sections and by transmission electron microscopy of liver, numerous long, thin, spiral-shaped bacteria were seen in the liver, spleen, cerebral ventricles, and within blood vessels in many organs. The organism was identified as a member of the Borrelia genus by sequence analysis of the PCR-amplified 16S rRNA gene. The most closely related species is B. hermsii, an agent of relapsing fever in humans in the western United States. This is the first report of a relapsing fever-related Borrelia in a wild bird.

Should sputum-negative presumptive TB patients be actively followed to identify missing cases in India?

PubMed

Waikar, S; Pathak, A; Ghule, V; Kapoor, A; Sagili, K; Babu, E R; Chadha, S

2017-12-21

Setting: Sputum smear microscopy, the primary diagnostic tool used for diagnosis of tuberculosis (TB) in India's Revised National TB Control Programme (RNTCP), has low sensitivity, resulting in a significant number of TB cases reported as sputum-negative. As the revised guidelines pose challenges in implementation, sputum-negative presumptive TB (SNPT) patients are subjected to 2 weeks of antibiotics, followed by chest X-ray (CXR), resulting in significant loss to care among these cases. Objective: To determine whether reducing delays in CXR would yield additional TB cases and reduce initial loss to follow-up for diagnosis among SNPT cases. Methods: In an ongoing intervention in five districts of Maharashtra, SNPT patients were offered upfront CXR. Results: Of 119 male and 116 female SNPT patients with a mean age of 45 years who were tested by CXR, 32 (14%) were reported with CXR suggestive of TB. Administering upfront CXR in SNPT patients yielded twice as many additional cases, doubling the proportion of cases detected among all those tested as against administering CXR 2 weeks after smear examination. Conclusion: Our interventional study showed that the yield of TB cases was significantly greater when upfront CXR examination was undertaken without waiting for a 2-week antibiotic trial.

Perceptions and concerns of women undergoing Pap smear examination in a tertiary care hospital of India.

PubMed

Tiwari, A; Kishore, J; Tiwari, A

2011-01-01

Cervical cancer is one of the major causes of deaths due to cancer among women in India. Pap smear is one of the best methods to detect early changes in cervix. However, there is lack of data on awareness level of women about Pap smear and various risk factors for cervical cancer. To study the awareness about various risk factors for cervical cancer, health-seeking behavior and hygienic practices among women and to assess the distress experienced by these women before the Pap smear examination. This cross-sectional study was carried out on women coming for a Pap smear examination in a tertiary teaching hospital in New Delhi. A pretested interview schedule was used to get information after obtaining their informed consent. Fifty-seven percent stated that they did not consult a doctor when they noticed the symptoms the first time. Sixty-one percent did not know what a cervical cancer is and a same percentage of women did not know what a Pap smear examination was. Older age group, Muslim and literate women had higher number of abnormal Pap smear results. Women who reported being stressed in their lives had higher number of abnormal smears as compared to women who claimed to lead a stress free life. Poor hygienic practices among these women from urban areas were also associated with abnormal Pap smear results. The study concluded that factors such as poor awareness, shyness, poor hygiene, and old age could be responsible for abnormal Pap smears and this needs special attention in cancer prevention activities of the government.

Comparison of Pap smear screening results between Akha hill tribe and urban women in Chiang Rai province, Thailand.

PubMed

Kritpetcharat, Onanong; Wutichouy, Wiwat; Sirijaichingkul, Suchat; Kritpetcharat, Panutas

2012-01-01

Cervical cancer is an important woman's health problems worldwide, especially in low socio-economic countries. The aim of this study was to compare the Pap smear screening results between Akha hill tribe and urban women who live in Chiang Rai province, Thailand. Screening was conducted for 1,100 Akha women and 1,100 urban women who came to have the Pap smear at Chiangrai Prachanukroh Hospital and 1 private cytology laboratory from January to June 2008. The demographic characteristics and factors related to abnormal Pap smears of these women were gathered using closed model questionnaires. Abnormal Rap smears were defined according to the Bethesda 2001 system. The results showed that the prevalence of abnormal Pap smears was 12.2% in Akha women and 4.5% in urban women. The highest prevalence of Pap abnormalities was found in the 41-50 years age group in both populations (4.5% in Akha and 1.7% in urban women). In both populations, abnormal Pap smears were found in <21 years age groups. From the questionnaires, the possible risk factors related to the higher prevalence of abnormal Pap smears in Akha women were early age at marriage (≤17 years), high frequency pregnacies and high parity and no/low education level. In conclusion, cervical cancer control by education and early detection by Pap smear screening is necessary for hill tribe women. More Pap smear screening service units should be set to improve the coverage for the risk group women who got married in young age, especial in ethnic groups.

[Abnormalities in cervical smears stored in plastic bags: potential cause of false negatives].

PubMed

Ortega-González, Patricia; González-Bravo, Margarita S; Jiménez-Muñoz-Ledo, Gustavo; Macías, Alejandro E

2011-01-01

Cervical smear is the most economic and efficient diagnostic tool for the screening of cervical cancer. However, since plastic bags have been used in Guanajuato to transport and store smears, we have observed cytological abnormalities which difficult the diagnosis and lead to false negatives. To describe those abnormalities. Out of 340 women registered in a primary care center in Mexico, 68 were selected through systematic random sampling during 2007. A cervical smear was obtained and placed on two slides. The first sample was allowed to dry but the second one was placed into the plastic bag immediately after fixation. After 15 days all the smears were stained with the Papanicolaou technique. A certified pathologist, blinded about the variable of study, interpreted the samples according to the Bethesda system, and evaluated the presence of necrosis, edema, holes, and opportunistic microorganisms. Of the 68 smears exposed to a humid storage, 36 (53%) were inadequate for diagnosis (Fisher's exact probability < 0.001). From them, 36 (53%) had holes or lagoons, 34 (50%) had edema, 31 (46%) had necrosis, and 15 (22%) had fungus. On the other hand, the 68 dried cervical smears were all adequate for diagnosis and none had the changes or cytological abnormalities. The humid transport and storage of cervical smears produced abnormalities in the normal morphology that could lead to false negative results. The guideline for the handling of cervical smears must stress the importance of allowing the smears to dry completely after fixation and before storing them in plastic bags.

Evaluation of final irrigation regimens with maleic acid for smear layer removal and wettability of root canal sealer.

PubMed

Ballal, Nidambur Vasudev; Ferrer-Luque, Carmen Maria; Sona, Mrunali; Prabhu, K Narayan; Arias-Moliz, Teresa; Baca, Pilar

2018-04-01

To evaluate the smear layer removal and wettability of AH Plus sealer on root canal dentin treated with MA (maleic acid), MA + CTR (cetrimide) and MA + CTR + CHX (chlorhexidine) as final irrigating regimens. For smear layer removal, 40 teeth were instrumented to size F4 and divided into four groups: (1) 7% MA, (2) 7% MA + 0.2% CTR, (3) 7% MA + 0.2% CTR + 2% CHX, (4) distilled water (control). After irrigation, teeth were subjected to SEM analysis. For contact angle analysis, 20 teeth were split longitudinally and divided into four groups similar to smear layer analysis. AH plus sealer was placed on each specimen and contact angle was analysed. In both smear layer (p = .393) and contact angle analysis (p = .961), there was no significant difference between the groups MA and MA + CTR. However, MA + CTR + CHX removed smear layer less effectively (p = .023) and increased the contact angle of the sealer (p = .005). In smear layer analysis, specimens in negative control group were heavily smeared. In case of contact angle analysis, samples in the control group had least contact angle. MA alone or in combination with CTR removed smear layer effectively and increased the wettability of AH plus sealer to root canal dentin.

Symptomatic candidiasis: Using self sampled vaginal smears to establish the presence of Candida, lactobacilli, and Gardnerella vaginalis.

PubMed

Engberts, M K; Boon, M E; van Haaften, M; Heintz, A P M

2007-10-01

In a prospective cohort study, 10 symptomatic women with recurrent vulvovaginal candidiasis were taught how to prepare vaginal smears of their own vaginal fluids on days 7, 14, 21, and 28. The 40 smears were stained with the PAS-method and examined by three different cytopathologists for presence of Candida. Thereafter, the smears were restained with Giemsa-stain to determine presence of lactobacilli, Gardnerella vaginalis ("clue cells") and neutrophils. All three cytopathologists unequivocally established Candida blastospores and (pseudo)hyphae in 27 out of the 40 PAS-stained vaginal smears, whereas in the remaining 13 smears Candida was not found. All 10 patients had Candida in their smears during the second half of their menstrual cycle.Self sampled smears prove to be reliable for establishing the presence of Candida in symptomatic patients with candidiasis. Candida is associated with a lactobacillus-predominated vaginal flora, but with the absence of Gardnerella vaginalis. Further studies may be directed towards the interaction between the various members of the vaginal flora. This study should open molecular methodology for determining the possible interactions of lactobacilli and Candida. (c) 2007 Wiley-Liss, Inc.

Motorcycle helmets: What about their coating?

PubMed

Schnegg, Michaël; Massonnet, Geneviève; Gueissaz, Line

2015-07-01

In traffic accidents involving motorcycles, paint traces can be transferred from the rider's helmet or smeared onto its surface. These traces are usually in the form of chips or smears and are frequently collected for comparison purposes. This research investigates the physical and chemical characteristics of the coatings found on motorcycles helmets. An evaluation of the similarities between helmet and automotive coating systems was also performed.Twenty-seven helmet coatings from 15 different brands and 22 models were considered. One sample per helmet was collected and observed using optical microscopy. FTIR spectroscopy was then used and seven replicate measurements per layer were carried out to study the variability of each coating system (intravariability). Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA) were also performed on the infrared spectra of the clearcoats and basecoats of the data set. The most common systems were composed of two or three layers, consistently involving a clearcoat and basecoat. The coating systems of helmets with composite shells systematically contained a minimum of three layers. FTIR spectroscopy results showed that acrylic urethane and alkyd urethane were the most frequent binders used for clearcoats and basecoats. A high proportion of the coatings were differentiated (more than 95%) based on microscopic examinations. The chemical and physical characteristics of the coatings allowed the differentiation of all but one pair of helmets of the same brand, model and color. Chemometrics (PCA and HCA) corroborated classification based on visual comparisons of the spectra and allowed the study of the whole data set at once (i.e., all spectra of the same layer). Thus, the intravariability of each helmet and its proximity to the others (intervariability) could be more readily assessed. It was also possible to determine the most discriminative chemical variables based on the study of the PCA loadings. Chemometrics could therefore be used as a complementary decision-making tool when many spectra and replicates have to be taken into account. Similarities between automotive and helmet coating systems were highlighted, in particular with regard to automotive coating systems on plastic substrates (microscopy and FTIR). However, the primer layer of helmet coatings was shown to differ from the automotive primer. If the paint trace contains this layer, the risk of misclassification (i.e., helmet versus vehicle) is reduced. Nevertheless, a paint examiner should pay close attention to these similarities when analyzing paint traces, especially regarding smears or paint chips presenting an incomplete layer system. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

The Tuberculosis Cascade of Care in India’s Public Sector: A Systematic Review and Meta-analysis

PubMed Central

Satyanarayana, Srinath; Pai, Madhukar; Thomas, Beena E.; Chadha, Vineet K.; Swaminathan, Soumya; Mayer, Kenneth H.

2016-01-01

Background India has 23% of the global burden of active tuberculosis (TB) patients and 27% of the world’s “missing” patients, which includes those who may not have received effective TB care and could potentially spread TB to others. The “cascade of care” is a useful model for visualizing deficiencies in case detection and retention in care, in order to prioritize interventions. Methods and Findings The care cascade constructed in this paper focuses on the Revised National TB Control Programme (RNTCP), which treats about half of India’s TB patients. We define the TB cascade as including the following patient populations: total prevalent active TB patients in India, TB patients who reach and undergo evaluation at RNTCP diagnostic facilities, patients successfully diagnosed with TB, patients who start treatment, patients retained to treatment completion, and patients who achieve 1-y recurrence-free survival. We estimate each step of the cascade for 2013 using data from two World Health Organization (WHO) reports (2014–2015), one WHO dataset (2015), and three RNTCP reports (2014–2016). In addition, we conduct three targeted systematic reviews of the scientific literature to identify 39 unique articles published from 2000–2015 that provide additional data on five indicators that help estimate different steps of the TB cascade. We construct separate care cascades for the overall population of patients with active TB and for patients with specific forms of TB—including new smear-positive, new smear-negative, retreatment smear-positive, and multidrug-resistant (MDR) TB. The WHO estimated that there were 2,700,000 (95%CI: 1,800,000–3,800,000) prevalent TB patients in India in 2013. Of these patients, we estimate that 1,938,027 (72%) TB patients were evaluated at RNTCP facilities; 1,629,906 (60%) were successfully diagnosed; 1,417,838 (53%) got registered for treatment; 1,221,764 (45%) completed treatment; and 1,049,237 (95%CI: 1,008,775–1,083,243), or 39%, of 2,700,000 TB patients achieved the optimal outcome of 1-y recurrence-free survival. The separate cascades for different forms of TB highlight different patterns of patient attrition. Pretreatment loss to follow-up of diagnosed patients and post-treatment TB recurrence were major points of attrition in the new smear-positive TB cascade. In the new smear-negative and MDR TB cascades, a substantial proportion of patients who were evaluated at RNTCP diagnostic facilities were not successfully diagnosed. Retreatment smear-positive and MDR TB patients had poorer treatment outcomes than the general TB population. Limitations of our analysis include the lack of available data on the cascade of care in the private sector and substantial uncertainty regarding the 1-y period prevalence of TB in India. Conclusions Increasing case detection is critical to improving outcomes in India’s TB cascade of care, especially for smear-negative and MDR TB patients. For new smear-positive patients, pretreatment loss to follow-up and post-treatment TB recurrence are considerable points of attrition that may contribute to ongoing TB transmission. Future multisite studies providing more accurate information on key steps in the public sector TB cascade and extension of this analysis to private sector patients may help to better target interventions and resources for TB control in India. PMID:27780217

The Tuberculosis Cascade of Care in India's Public Sector: A Systematic Review and Meta-analysis.

PubMed

Subbaraman, Ramnath; Nathavitharana, Ruvandhi R; Satyanarayana, Srinath; Pai, Madhukar; Thomas, Beena E; Chadha, Vineet K; Rade, Kiran; Swaminathan, Soumya; Mayer, Kenneth H

2016-10-01

India has 23% of the global burden of active tuberculosis (TB) patients and 27% of the world's "missing" patients, which includes those who may not have received effective TB care and could potentially spread TB to others. The "cascade of care" is a useful model for visualizing deficiencies in case detection and retention in care, in order to prioritize interventions. The care cascade constructed in this paper focuses on the Revised National TB Control Programme (RNTCP), which treats about half of India's TB patients. We define the TB cascade as including the following patient populations: total prevalent active TB patients in India, TB patients who reach and undergo evaluation at RNTCP diagnostic facilities, patients successfully diagnosed with TB, patients who start treatment, patients retained to treatment completion, and patients who achieve 1-y recurrence-free survival. We estimate each step of the cascade for 2013 using data from two World Health Organization (WHO) reports (2014-2015), one WHO dataset (2015), and three RNTCP reports (2014-2016). In addition, we conduct three targeted systematic reviews of the scientific literature to identify 39 unique articles published from 2000-2015 that provide additional data on five indicators that help estimate different steps of the TB cascade. We construct separate care cascades for the overall population of patients with active TB and for patients with specific forms of TB-including new smear-positive, new smear-negative, retreatment smear-positive, and multidrug-resistant (MDR) TB. The WHO estimated that there were 2,700,000 (95%CI: 1,800,000-3,800,000) prevalent TB patients in India in 2013. Of these patients, we estimate that 1,938,027 (72%) TB patients were evaluated at RNTCP facilities; 1,629,906 (60%) were successfully diagnosed; 1,417,838 (53%) got registered for treatment; 1,221,764 (45%) completed treatment; and 1,049,237 (95%CI: 1,008,775-1,083,243), or 39%, of 2,700,000 TB patients achieved the optimal outcome of 1-y recurrence-free survival. The separate cascades for different forms of TB highlight different patterns of patient attrition. Pretreatment loss to follow-up of diagnosed patients and post-treatment TB recurrence were major points of attrition in the new smear-positive TB cascade. In the new smear-negative and MDR TB cascades, a substantial proportion of patients who were evaluated at RNTCP diagnostic facilities were not successfully diagnosed. Retreatment smear-positive and MDR TB patients had poorer treatment outcomes than the general TB population. Limitations of our analysis include the lack of available data on the cascade of care in the private sector and substantial uncertainty regarding the 1-y period prevalence of TB in India. Increasing case detection is critical to improving outcomes in India's TB cascade of care, especially for smear-negative and MDR TB patients. For new smear-positive patients, pretreatment loss to follow-up and post-treatment TB recurrence are considerable points of attrition that may contribute to ongoing TB transmission. Future multisite studies providing more accurate information on key steps in the public sector TB cascade and extension of this analysis to private sector patients may help to better target interventions and resources for TB control in India.

Cervical cancer screening performed by a nurse. Evaluation in family practice.

PubMed Central

Thommasen, H.; Lenci, P.; Brake, I.; Anderson, G.

1996-01-01

OBJECTIVE: To determine whether a nurse practitioner could collect adequate Papanicolaou smear samples from the transformation zone of the cervix. DESIGN: A retrospective, descriptive study. SETTING: The Bella Coola Medical Clinic, a primary care facility located in the isolated, small village of Bella Coola, BC. PARTICIPANTS: All women who presented for Pap smears between July 1993 and June 1994. MAIN OUTCOME MEASURES: Endocervical or metaplastic cells in smear samples. RESULTS: All Pap smears performed in the Bella Coola Medical Clinic over 1 year by either the nurse or a member of the physician group (which includes family practice residents) were reviewed. Between July 1, 1993, and June 30, 1994, 149 Pap smears were done, 55 by the nurse and 94 by the physicians. All smears collected by the nurse practitioner showed endocervical or metaplastic cells. More than 90% of physician samples showed endocervical or metaplastic cells. CONCLUSIONS: A Canadian nurse practitioner can be trained to collect adequate Pap smears. PMID:8939319

Identification of Dekkera bruxellensis (Brettanomyces) from Wine by Fluorescence In Situ Hybridization Using Peptide Nucleic Acid Probes

PubMed Central

Stender, Henrik; Kurtzman, Cletus; Hyldig-Nielsen, Jens J.; Sørensen, Ditte; Broomer, Adam; Oliveira, Kenneth; Perry-O'Keefe, Heather; Sage, Andrew; Young, Barbara; Coull, James

2001-01-01

A new fluorescence in situ hybridization method using peptide nucleic acid (PNA) probes for identification of Brettanomyces is described. The test is based on fluorescein-labeled PNA probes targeting a species-specific sequence of the rRNA of Dekkera bruxellensis. The PNA probes were applied to smears of colonies, and results were interpreted by fluorescence microscopy. The results obtained from testing 127 different yeast strains, including 78 Brettanomyces isolates from wine, show that the spoilage organism Brettanomyces belongs to the species D. bruxellensis and that the new method is able to identify Brettanomyces (D. bruxellensis) with 100% sensitivity and 100% specificity. PMID:11157265

External quality assessment of malaria microscopy diagnosis in selected health facilities in Western Oromia, Ethiopia.

PubMed

Sori, Getachew; Zewdie, Olifan; Tadele, Geletta; Samuel, Abdi

2018-06-18

Accurate early diagnosis and prompt treatment are one of the key strategies to control and prevent malaria disease. External quality assessment is the most effective method for evaluation of the quality of malaria microscopy diagnosis. The aim of this study was to assess the quality of malaria microscopy diagnosis and its associated factors in selected public health facility laboratories in East Wollega Zone, Western Ethiopia. Facility-based cross-sectional study design was conducted in 30 randomly selected public health facility laboratories from November 2014 to January 2015 in East Wollega Zone, Western Ethiopia. Ten validated stained malaria panel slides with known Plasmodium species, developmental stage and parasite density were distributed. Data were captured; cleaned and analyzed using SPSS version 20 statistical software-multivariate logistic regressions and the agreement in reading between the peripheral diagnostic centers and the reference laboratory were done using kappa statistics. A total of 30 health facility laboratories were involved in the study and the overall quality of malaria microscopy diagnosis was poor (62.3%). The associated predictors of quality in this diagnosis were in-service training [(AOR = 16, 95% CI (1.3, 1.96)], smearing quality [(AOR = 24, 95% CI (1.8, 3.13)], staining quality [(AOR = 15, 95% CI (2.35, 8.61), parasite detection [(AOR = 9, 95% CI (1.1, 8.52)] and identification skills [(AOR = 8.6, 95% CI (1.21, 1.63)]. Eighteen (60%) of health facility laboratories had in-service trained laboratory professionals on malaria microscopy diagnosis. Overall quality of malaria microscopy diagnosis was poor and a significant gap in this service was observed that could impact on its diagnostic services.

Performance of a Novel Algorithm Using Automated Digital Microscopy for Diagnosing Tuberculosis.

PubMed

Ismail, Nazir A; Omar, Shaheed V; Lewis, James J; Dowdy, David W; Dreyer, Andries W; van der Meulen, Hermina; Nconjana, George; Clark, David A; Churchyard, Gavin J

2015-06-15

TBDx automated microscopy is a novel technology that processes digital microscopic images to identify acid-fast bacilli (AFB). Use of TBDx as part of a diagnostic algorithm could improve the diagnosis of tuberculosis (TB), but its performance characteristics have not yet been formally tested. To evaluate the performance of the TBDx automated microscopy system in algorithms for diagnosis of TB. Prospective samples from patients with presumed TB were processed in parallel with conventional smear microscopy, TBDx microscopy, and liquid culture. All TBDx-positive specimens were also tested with the Xpert MTB/RIF (GXP) assay. We evaluated the sensitivity and specificity of two algorithms-(1) TBDx-GXP (TBDx with positive specimens tested by Xpert MTB/RIF) and (2) TBDx alone-against the gold standard liquid media culture. Of 1,210 samples, 1,009 were eligible for evaluation, of which 109 were culture positive for Mycobacterium tuberculosis. The TBDx system identified 70 specimens (68 culture positive) as having 10 or more putative AFB (high positive) and 207 (19 culture positive) as having 1-9 putative AFB (low positive). An algorithm in which "low-positive" results on TBDx were confirmed by GXP had 78% sensitivity (85 of 109) and 99.8% specificity (889 of 900), requiring 21% (207 of 1,009) specimens to be processed by GXP. As a stand-alone test, a "high-positive" result on TBDx had 62% sensitivity and 99.7% specificity. TBDx used in diagnostic algorithms with GXP provided reasonable sensitivity and high specificity for active TB while dramatically reducing the number GXP tests performed. As a stand-alone microscopy system, its performance was equivalent to that of a highly experienced TB microscopist.

Locally smeared operator product expansions in scalar field theory

DOE PAGES

Monahan, Christopher; Orginos, Kostas

2015-04-01

We propose a new locally smeared operator product expansion to decompose non-local operators in terms of a basis of smeared operators. The smeared operator product expansion formally connects nonperturbative matrix elements determined numerically using lattice field theory to matrix elements of non-local operators in the continuum. These nonperturbative matrix elements do not suffer from power-divergent mixing on the lattice, which significantly complicates calculations of quantities such as the moments of parton distribution functions, provided the smearing scale is kept fixed in the continuum limit. The presence of this smearing scale complicates the connection to the Wilson coefficients of the standardmore » operator product expansion and requires the construction of a suitable formalism. We demonstrate the feasibility of our approach with examples in real scalar field theory.« less

Relationship Between Pelvic Organ Prolapse and Non-Human Papillomavirus Pap Smear Abnormalities.

PubMed

Menhaji, Kimia; Harvie, Heidi S; Cheston, Emily; Levin, Pamela J; Arya, Lily A; Andy, Uduak U

2017-07-13

The aim of this study was to determine the association between pelvic organ prolapse (POP) and non-human papillomavirus (HPV) Papanicolaou (Pap) smear abnormalities. This was a retrospective cohort study of women aged 40 to 70 years who presented for consultation at our institution between 2010 and 2015 and had results of a Pap smear and HPV test available within 5 years of their visit. We extracted demographic information, medical and social history, Pap smear, and HPV results from the electronic medical record. Associations between the presence of POP and non-HPV Pap smear abnormalities were estimated using univariable and multivariable analyses. We reviewed 1590 charts and excluded 980 women, leaving 610 women in the study: 183 with POP and 427 without POP. Women with POP were significantly older (58.2 ± 7.2 vs 55.6 ± 6.6, P < 0.01) and more likely to have a remote (>10 year) history of abnormal Pap smear (24.0% vs 14.8%, P < 0.01). The rate of non-HPV-associated abnormal Pap smears was higher in the POP group than in the non-POP group (12/183 [6.6%] vs 12/427 [2.8%], P = 0.029). In the POP group, the rate of non-HPV Pap smear abnormality was significantly associated with increasing prolapse stage (stage 1: 0/16 [0%], stage 2: 5/77 [6.5%], stage 3: 3/73 [4.1%], stage 4: 4/17 [23.5%]; P = 0.02). After controlling for age and remote history of abnormal Pap smear, the odds ratio for non-HPV Pap smear abnormalities in the POP group remained significant (2.49; 95% confidence interval, 1.08-5.79). Human papillomavirus-negative Pap smear abnormalities may be related to POP. Our findings have important implications for surgeons seeking to leave the cervix in situ in women with POP.

Performance evaluation of automated urine microscopy as a rapid, non-invasive approach for the diagnosis of non-gonococcal urethritis.

PubMed

Pond, Marcus J; Nori, Achyuta V; Patel, Sheel; Laing, Ken; Ajayi, Margarita; Copas, Andrew J; Butcher, Philip D; Hay, Phillip; Sadiq, Syed Tariq

2015-05-01

Gram-stained urethral smear (GSUS), the standard point-of-care test for non-gonococcal urethritis (NGU) is operator dependent and poorly specific. The performance of rapid automated urine flow cytometry (AUFC) of first void urine (FVU) white cell counts (UWCC) for predicting Mycoplasma genitalium and Chlamydia trachomatis urethral infections was assessed and its application to asymptomatic infection was evaluated. Receiver operating characteristic curve analysis, determining FVU-UWCC threshold for predicting M. genitalium or C. trachomatis infection was performed on 208 'training' samples from symptomatic patients and subsequently validated using 228 additional FVUs obtained from prospective unselected patients. An optimal diagnostic threshold of >29 UWC/µL gave sensitivities and specificities for either infection of 81.5% (95% CI 65.1% to 91.6%) and 85.8% (79.5% to 90.4%), respectively, compared with 86.8% (71.1% to 95%) and 64.7% (56.9% to 71.7%), respectively, for GSUS, using the training set samples. FVU-UWCC demonstrated sensitivities and specificities of 69.2% (95% CI 48.1% to 84.9%) and 92% (87.2% to 95.2%), respectively, when using validation samples. In asymptomatic patients where GSUS was not used, AUFC would have enabled more infections to be detected compared with clinical considerations only (71.4% vs 28.6%; p=0.03). The correlation between UWCC and bacterial load was stronger for M. genitalium compared with C. trachomatis (τ=0.426, p≤0.001 vs τ=0.295, p=0.022, respectively). AUFC offers improved specificity over microscopy for predicting C. trachomatis or M. genitalium infection. Universal AUFC may enable non-invasive diagnosis of asymptomatic NGU at the PoC. The degree of urethral inflammation exhibits a stronger association with pathogen load for M. genitalium compared with C. trachomatis. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

Intermediate-grade squamous intraepithelial lesion may be a valid diagnostic/interpretive category.

PubMed

Ravinsky, Esther; Baker, Patricia

2009-02-01

We undertook this study to assess the characteristics of smears with features intermediate between high-grade squamous intraepithelial lesion (HSIL) and low-grade squamous intraepithelial lesion (ISIL). We also wanted to determine how these smears correlate with high risk biopsy diagnosis and to compare this with the biopsy correlation of LSIL and HSIL. Seventy-four squamous intraepithelial lesion (SIL) smears were identified as intermediate-grade SIL smears taken at colposcopy in a 1 year period. They were correlated with concurrent colposcopically guided biopsies. Thirty-five percent of cases with intermediate-grade SIL smears had a biopsy diagnosis of moderate dysplasia or higher as compared with 12% for LSIL 74% for HSIL. This confirmed our hypothesis that intermediate-grade SIL smears have a rate of biopsy diagnosis of moderate dysplasia or higher intermediate to that of LSIL and HSIL.

Defining the Needs for Next Generation Assays for Tuberculosis

PubMed Central

Denkinger, Claudia M.; Kik, Sandra V.; Cirillo, Daniela Maria; Casenghi, Martina; Shinnick, Thomas; Weyer, Karin; Gilpin, Chris; Boehme, Catharina C.; Schito, Marco; Kimerling, Michael; Pai, Madhukar

2015-01-01

To accelerate the fight against tuberculosis, major diagnostic challenges need to be addressed urgently. Post-2015 targets are unlikely to be met without the use of novel diagnostics that are more accurate and can be used closer to where patients first seek care in affordable diagnostic algorithms. This article describes the efforts by the stakeholder community that led to the identification of the high-priority diagnostic needs in tuberculosis. Subsequently target product profiles for the high-priority diagnostic needs were developed and reviewed in a World Health Organization (WHO)-led consensus meeting. The high-priority diagnostic needs included (1) a sputum-based replacement test for smear-microscopy; (2) a non-sputum-based biomarker test for all forms of tuberculosis, ideally suitable for use at levels below microscopy centers; (3) a simple, low cost triage test for use by first-contact care providers as a rule-out test, ideally suitable for use by community health workers; and (4) a rapid drug susceptibility test for use at the microscopy center level. The developed target product profiles, along with complimentary work presented in this supplement, will help to facilitate the interaction between the tuberculosis community and the diagnostics industry with the goal to lead the way toward the post-2015 global tuberculosis targets. PMID:25765104

Clinical value of whole-blood interferon-gamma assay in patients with suspected pulmonary tuberculosis and AFB smear- and polymerase chain reaction-negative bronchial aspirates.

PubMed

Lee, Jaehee; Lee, Shin Yup; Yoo, Seung Soo; Cha, Seung Ick; Won, Dong Il; Park, Jae Yong; Lee, Won-Kil; Kim, Chang Ho

2012-07-01

Combining a polymerase chain reaction (PCR) test with bronchoscopy is frequently performed to allow a rapid diagnosis of smear-negative pulmonary tuberculosis (PTB). However, limited data are available concerning clinical judgment in patients with suspected PTB and AFB smear- and PCR-negative bronchial aspirates (BA). The present study evaluated the usefulness of whole-blood QuantiFERON-TB Gold In-Tube (QFT) testing in these patients. Of 166 patients with suspected PTB who had undergone bronchoscopy because of smear-negative sputum or inadequate sputum production, 93 (56%) were diagnosed with culture-positive PTB. Seventy-four patients were either AFB smear- or PCR-positive. In the 75 patients whose BA AFB smear and PCR results were both negative, 19 were finally diagnosed with PTB by culture. The QFT test had a negative predictive value of 91% for PTB. The QFT test may be useful for excluding PTB in patients with suspected PTB whose BA AFB smear and PCR results are both negative. Copyright © 2012 Elsevier Inc. All rights reserved.

[Squamous intraepithlelial lesions in women's lupus].

PubMed

Mercado, Ulises

2009-09-01

previous studies have suggested an association between systemic lupus erythematosus (SLE), abnormal cervical smears and squamous intraepithelial lesions (SIL), but the underlying cause of this association is not well defined. To review Pap smear and biopsy results in SLE women compared with healthy controls and to identify traditional risk factors associated with abnormal smears. Retrospective cohort study. Sixty-two SLE patients and 1719 controls were included. Women with abnormal Pap smear results were referred to colposcopic biopsy histology. Information on traditional risk factors was obtained. Among SLE patients, activity score (Mex-SLEDAI) and immunosuppressive drugs exposure also were determined. Fourteen (22%) out of 62 patients had abnormal Pap smear results, compared with 120 (7%) out of 1719 controls. Thirteen (92%) out of 14 patients showed SIL by histological examination, compared with 27 (22%) out of 120 controls. Two SLE women had condylomata. There were no significant differences in the use of immunosuppressive agents among SLE patients with and without SIL. Abnormal Pap smears, SIL by biopsy and changes in the sexual behavioral were more common among SLE patients than in controls. The immunosuppressive drug exposure was not associated with abnormal Pap smears.

The large deviation function for entropy production: the optimal trajectory and the role of fluctuations

NASA Astrophysics Data System (ADS)

Speck, Thomas; Engel, Andreas; Seifert, Udo

2012-12-01

We study the large deviation function for the entropy production rate in two driven one-dimensional systems: the asymmetric random walk on a discrete lattice and Brownian motion in a continuous periodic potential. We compare two approaches: using the Donsker-Varadhan theory and using the Freidlin-Wentzell theory. We show that the wings of the large deviation function are dominated by a single optimal trajectory: either in the forward direction (positive rate) or in the backward direction (negative rate). The joining of the two branches at zero entropy production implies a non-differentiability and thus the appearance of a ‘kink’. However, around zero entropy production, many trajectories contribute and thus the ‘kink’ is smeared out.

Development of a Nonequilibrium Radiative Heating Prediction Method for Coupled Flowfield Solutions

NASA Technical Reports Server (NTRS)

Hartung, Lin C.

1991-01-01

A method for predicting radiative heating and coupling effects in nonequilibrium flow-fields has been developed. The method resolves atomic lines with a minimum number of spectral points, and treats molecular radiation using the smeared band approximation. To further minimize computational time, the calculation is performed on an optimized spectrum, which is computed for each flow condition to enhance spectral resolution. Additional time savings are obtained by performing the radiation calculation on a subgrid optimally selected for accuracy. Representative results from the new method are compared to previous work to demonstrate that the speedup does not cause a loss of accuracy and is sufficient to make coupled solutions practical. The method is found to be a useful tool for studies of nonequilibrium flows.

Market assessment of tuberculosis diagnostics in Brazil in 2012.

PubMed

2014-01-01

Improved diagnostics for the diagnosis of tuberculosis (TB) are urgently needed. However, test developers and investors require market size data to support new product development. This study assessed the served available market for TB diagnostics in Brazil in 2012 and the market segmentation in the public and private sectors. Data were collected on test volumes done in the public and private sectors for the diagnosis of latent and active TB, drug susceptibility testing and treatment follow-up. Tests included were tuberculin skin tests, interferon-gamma releases assays, smear microscopy, solid and liquid cultures, nucleic acid amplification tests and phenotypic drug susceptibility tests. The data were collected by means of an electronic survey via the Brazilian State laboratories and from sales information provided by manufacturers. Test costs for the public sector were calculated using a components approach, while costs for the private sector were based on prices paid by patients. The overall market value (expenditure) for the entire country was calculated using the public sector test costs. During 2012, an estimated total of 2.4 million TB diagnostic tests were done in Brazil, resulting in an estimated overall market value of USD 17.2 million. The public sector accounted for 91% of the test volumes and 88% of the market value. Smear microscopy was the most commonly test (n = 1.3 million; 55% of total) at an estimated value of USD 3.7 million. Culture overall (n = 302,761) represented 13% of test volumes and 40% (USD 6.9 million) of the market value. On average, USD 208 was spent on TB diagnostics for every notified TB patient in Brazil, in 2012. The TB diagnostics market value in Brazil in 2012 was over USD 17 million. These study results will help test developers to understand the current and potential market for replacement or add-on diagnostic technologies.

[Evaluation of the analyzer of hematology Beckman Coulter® HmX™ in the university hospital of Oran].

PubMed

Zmouli, N; Moulasserdoun, K; Seghier, F

2013-11-01

The choice of an automaton of haematology is a determining stage, which has to take into account at the same time the quality of the results and the economic imperatives: workload, structure and organization of the laboratory. [corrected] It is in this spirit that we estimated during a period of 3 months the analyzer of haematology: the HmX™ Coulter with boatman of samples of the company Beckman. This automaton realizes the blood numeration, the formula leukocytic and the reticulocyte count. At first, we estimated the appropriate characteristics of device. Secondly, we estimated the relevance, the sensibility and the specificity of the alarms by comparing with the reference method, which is the optical microscopy. For that purpose, 125 blood smears resulting from service of haematology and from resuscitation were examined in optical microscopy. The technical tests were realized according to the recommendations of the International committee for evaluation of automatons of haematology. The analytical performances were satisfactory in particular the big interval of linearity and the absence of contamination. As regards the evaluation of the alarms system: rate of rejection is 63%, the sensibility 86%, the specificity 70%, the positive predictive value 80%, the negative predictive value 78% and the efficiency 80%. The alarms myelaemia and atypical lymphocytes were never sources of false negatives. The alarms erythroblasts and platelet aggregates did not engendered positive forgery. The blast cell alarm was responsible for a single case of false negative. The faithfulness of automaton is satisfactory: the absence of contamination, the big interval of linearity for the leukocytes, the red blood cells and the platelets as well as a good relevance of the alarms with regard to the anomalies found on the peripheral blood smear. From the user-friendliness and practicability point of view, the HmX™ Coulter was deeply appreciated. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Comparison of TB-LAMP, GeneXpert MTB/RIF and culture for diagnosis of pulmonary tuberculosis in The Gambia.

PubMed

Bojang, Adama L; Mendy, Francis S; Tientcheu, Leopold D; Otu, Jacob; Antonio, Martin; Kampmann, Beate; Agbla, Schadrac; Sutherland, Jayne S

2016-03-01

Diagnosis of tuberculosis (TB) remains difficult, particularly in resource-limited settings. The development of nucleic acid-based tests for detection of Mycobacterium tuberculosis complex (MTBC) has significantly increased sensitivity compared to conventional smear microscopy and provides results within a matter of hours compared to weeks for the current gold-standard, liquid culture. In this study we performed side-by-side comparison of mycobacterial detection assays on sputum samples from 285 subjects presenting with symptoms suggestive of TB in The Gambia and a cross-sectional cohort of 156 confirmed TB patients with a median of 2 months of treatment. A novel assay, Loop-Mediated Amplification test for TB (TB-LAMP), was compared to smear microscopy, MGIT culture and GeneXpert MTB/RIF for all samples. When culture was used as the reference standard, we found an overall sensitivity for TB-LAMP of 99% (95% CI: 94.5-99.8) and specificity of 94% (95% CI: 89.3-96.7). When latent class analysis was performed, TB-LAMP had 98.6% (95% CI: 95.9-100) sensitivity and 99% (95% CI: 98.2-100) specificity compared to 91.1% (95% CI: 86.1-96) sensitivity and 100% (95% CI: 98.2-100) specificity for MGIT culture. GeneXpert had the highest sensitivity 99.1% (95% CI: 97.1-100) but the lowest specificity 96% (95% CI: 92.6-98.3). Both TB-LAMP and GeneXpert showed high sensitivity and specificity regardless of age or strain of infection. Our findings show the diagnostic utility of both GeneXpert and TB-LAMP in The Gambia. Whilst TB-LAMP requires less infrastructure, it is unable to detect drug-resistant patterns and therefore would be most suitable as a screening test for new TB cases in peripheral health clinics. Copyright © 2015 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

Diagnostic and prognostic utility of an inexpensive rapid on site malaria diagnostic test (ParaHIT f) among ethnic tribal population in areas of high, low and no transmission in central India.

PubMed

Singh, Neeru; Mishra, A K; Shukla, M M; Chand, S K; Bharti, Praveen Kumar

2005-06-21

Malaria presents a diagnostic challenge in most tropical countries. Rapid detection of the malaria parasite and early treatment of infection still remain the most important goals of disease management. Therefore, performance characteristics of the new indigenous ParaHIT f test (Span diagnostic Ltd, Surat, India) was determined among ethnic tribal population in four districts of different transmission potential in central India to assess whether this rapid diagnostic test (RDT) could be widely applied as a diagnostic tool to control malaria. Beyond diagnosis, the logical utilization of RDTs is to monitor treatment outcome. A finger prick blood sample was collected from each clinically suspected case of malaria to prepare blood smear and for testing with the RDT after taking informed consent. The blood smears were read by an experienced technician blinded to the RDT results and clinical status of the subjects. The figures for specificity, sensitivity, accuracy and predictive values were calculated using microscopy as gold standard. The prevalence of malaria infection estimated by RDT in parallel with microscopy provide evidence of the type of high, low or no transmission in the study area. Analysis revealed (pooled data of all four epidemiological settings) that overall sensitivity, specificity and accuracy of the RDT were >90% in areas of different endemicity. While, RDT is useful to confirm the diagnosis of new symptomatic cases of suspected P. falciparum infection, the persistence of parasite antigen leading to false positives even after clearance of asexual parasitaemia has limited its utility as a prognostic tool. The study showed that the ParaHIT f test was easy to use, reliable and cheap. Thus this RDT is an appropriate test for the use in the field by paramedical staff when laboratory facilities are not available and thus likely to contribute greatly to an effective control of malaria in resource poor countries.

A comparative scanning electron microscopy study between hand instrument, ultrasonic scaling and erbium doped:Yttirum aluminum garnet laser on root surface: A morphological and thermal analysis

PubMed Central

Mishra, Mitul Kumar; Prakash, Shobha

2013-01-01

Background and Objectives: Scaling and root planing is one of the most commonly used procedures for the treatment of periodontal diseases. Removal of calculus using conventional hand instruments is incomplete and rather time consuming. In search of more efficient and less difficult instrumentation, investigators have proposed lasers as an alternative or as adjuncts to scaling and root planing. Hence, the purpose of the present study was to evaluate the effectiveness of erbium doped: Yttirum aluminum garnet (Er:YAG) laser scaling and root planing alone or as an adjunct to hand and ultrasonic instrumentation. Subjects and Methods: A total of 75 freshly extracted periodontally involved single rooted teeth were collected. Teeth were randomly divided into five treatment groups having 15 teeth each: Hand scaling only, ultrasonic scaling only, Er:YAG laser scaling only, hand scaling + Er:YAG laser scaling and ultrasonic scaling + Er:YAG laser scaling. Specimens were subjected to scanning electron microscopy and photographs were evaluated by three examiners who were blinded to the study. Parameters included were remaining calculus index, loss of tooth substance index, roughness loss of tooth substance index, presence or absence of smear layer, thermal damage and any other morphological damage. Results: Er:YAG laser treated specimens showed similar effectiveness in calculus removal to the other test groups whereas tooth substance loss and tooth surface roughness was more on comparison with other groups. Ultrasonic treated specimens showed better results as compared to other groups with different parameters. However, smear layer presence was seen more with hand and ultrasonic groups. Very few laser treated specimens showed thermal damage and morphological change. Interpretation and Conclusion: In our study, ultrasonic scaling specimen have shown root surface clean and practically unaltered. On the other hand, hand instrument have produced a plane surface, but removed more tooth structure. The laser treated specimens showed rough surfaces without much residual deposit or any other sign of morphological change. PMID:24015009

Market Assessment of Tuberculosis Diagnostics in Brazil in 2012

PubMed Central

2014-01-01

Background Improved diagnostics for the diagnosis of tuberculosis (TB) are urgently needed. However, test developers and investors require market size data to support new product development. This study assessed the served available market for TB diagnostics in Brazil in 2012 and the market segmentation in the public and private sectors. Methods Data were collected on test volumes done in the public and private sectors for the diagnosis of latent and active TB, drug susceptibility testing and treatment follow-up. Tests included were tuberculin skin tests, interferon-gamma releases assays, smear microscopy, solid and liquid cultures, nucleic acid amplification tests and phenotypic drug susceptibility tests. The data were collected by means of an electronic survey via the Brazilian State laboratories and from sales information provided by manufacturers. Test costs for the public sector were calculated using a components approach, while costs for the private sector were based on prices paid by patients. The overall market value (expenditure) for the entire country was calculated using the public sector test costs. Results During 2012, an estimated total of 2.4 million TB diagnostic tests were done in Brazil, resulting in an estimated overall market value of USD 17.2 million. The public sector accounted for 91% of the test volumes and 88% of the market value. Smear microscopy was the most commonly test (n = 1.3 million; 55% of total) at an estimated value of USD 3.7 million. Culture overall (n = 302,761) represented 13% of test volumes and 40% (USD 6.9 million) of the market value. On average, USD 208 was spent on TB diagnostics for every notified TB patient in Brazil, in 2012. Conclusion The TB diagnostics market value in Brazil in 2012 was over USD 17 million. These study results will help test developers to understand the current and potential market for replacement or add-on diagnostic technologies. PMID:25099237

Stool guaiac test

MedlinePlus

... guaiac test; gFOBT; Guaiac smear test; Fecal occult blood test - guaiac smear; Stool occult blood test - guaiac smear ... This test detects blood in the digestive tract. It may be done if: You are being screened or tested for colon cancer You ...

Blood Smear: MedlinePlus Lab Test Information

MedlinePlus

... Test Information → Blood Smear URL of this page: https://medlineplus.gov/labtests/bloodsmear.html Blood Smear To ... May 26]; 353(5): 498–507. Available from: http://www.nejm.org/doi/full/10.1056/NEJMra043442 ...

Evaluation and optimization of the extended information process unit (E-IPU) validation module integrating the sysmex flag systems and the recommendations of the French-speaking cellular hematology group (GFHC).

PubMed

Cornet, Edouard; Mullier, François; Despas, Noemie; Jacqmin, Hugues; Geara, Carole; Boubaya, Marouane; Chatelain, Bernard; Troussard, Xavier

2016-10-01

The French-Speaking Cellular Haematology Group (GFHC) recently published criteria for microscopic analysis of a blood smears when a hemogram is requested. In order to evaluate and improve these recommendations using an XN (Sysmex) analyzer, we assessed 31,836 samples categorized into two sub-groups of patients either receiving or not receiving care in the clinical hematology/oncology departments of two university hospitals. By combining the manufacturer's recommendations and the GFHC recommendations, 21.3% of samples had a positive review flag in phase 1 of our study (17,991 samples). In phase 2 (13,845 samples), increasing the immature granulocytes (IG) percentage from 5-10% as a review trigger threshold, and ignoring slides with isolated flags 'PLT HIGH' (thrombocytosis) or 'MCV LOW' (microcytosis) or 'Blast/Abn Lymph and Atypical Lymph' (blast cells/abnormal lymphocytes and atypical lymphocytes) (in the absence of abnormal cells on a previous blood smear within 72 h), enabled us to significantly reduce the number of slides reviewed from 21.3-15.0% (p < 0.0001), without loss of clinical value. This decrease occurred in both sub-groups (hematology 48.7-38.0%, non-hematology 18.3-11.7%, p < 0.0001). In conclusion, the application of the GFHC criteria adapted to XN analyzers has enabled us to optimize the hematology laboratory processes, and thus reduce the production costs and the turnaround time of hemogram results.

Evaluation of case management of uncomplicated malaria in Haiti: a national health facility survey, 2012.

PubMed

Landman, Keren Z; Jean, Samuel E; Existe, Alexandre; Akom, Eniko E; Chang, Michelle A; Lemoine, Jean Frantz; Mace, Kimberly E

2015-10-09

Malaria is a public health concern in Haiti, although there are limited data on its burden and case management. National malaria guidelines updated in 2012 recommend treatment with chloroquine and primaquine. In December 2012, a nationally-representative cross-sectional survey of health facilities (HFs) was conducted to determine malaria prevalence among febrile outpatients and malaria case management quality at baseline before scale-up of diagnostics and case management training. Among all 833 HFs nationwide, 30 were selected randomly, in proportion to total HFs per region, for 2-day evaluations. Survey teams inventoried HF material and human resources. Outpatients of all ages were screened for temperature >37.5 °C or history of fever; those without severe symptoms were consented and enrolled. Providers evaluated and treated enrolled patients according to HF standards; the survey teams documented provider-ordered diagnostic tests and treatment decisions. Facility-based test results [microscopy and malaria rapid diagnostic tests (RDTs)] were collected from HF laboratories. Blood smears for gold-standard microscopy, and dried blood spots for polymerase chain reaction (PCR) were obtained. Malaria diagnostic capacity, defined as completing a test for an enrolled patient or having adequate resources for RDTs or microscopy, was present in 11 (37 %) HFs. Among 459 outpatients screened, 257 (56 %) were febrile, of which 193 (75 %) were eligible, and 153 (80 %) were enrolled. Among 39 patients with facility-level malaria test results available on the survey day, 11 (28 %) were positive, of whom 6 (55 %) were treated with an anti-malarial. Twenty-seven (95 %) of the 28 patients testing negative were not treated with an anti-malarial. Of 114 patients without test results available, 35 (31 %) were presumptively treated for malaria. Altogether, 42 patients were treated with an anti-malarial, one (2 %) according to Haiti's 2012 guidelines. Of 140 gold-standard smears, none were positive, although one patient tested positive by PCR, a more sensitive technique. The national prevalence of malaria among febrile outpatients is estimated to be 0.5 % (95 % confidence interval 0-1.7 %). Malaria is an uncommon cause of fever in Haitian outpatients, and limited, often inaccurate, diagnostic capacity at baseline contributes to over diagnosis. Scale-up of diagnostics and training on new guidelines should improve malaria diagnosis and treatment in Haiti.

Use of the Xpert(®) MTB/RIF assay for diagnosing pulmonary tuberculosis comorbidity and multidrug-resistant TB in obstetrics and gynaecology inpatient wards at the University Teaching Hospital, Lusaka, Zambia.

PubMed

Bates, Matthew; Ahmed, Yusuf; Chilukutu, Lophina; Tembo, John; Cheelo, Busiku; Sinyangwe, Sylvester; Kapata, Nathan; Maeurer, Markus; O'Grady, Justin; Mwaba, Peter; Zumla, Alimuddin

2013-09-01

In high-tuberculosis (TB)-endemic countries, comorbidity of pulmonary TB in hospitalised patients with non-communicable diseases is well documented. In this study, we evaluated the use of the Xpert(®) MTB/RIF assay for the detection of concomitant pulmonary TB in patients admitted to the University Teaching Hospital, Lusaka, Zambia, with a primary obstetric or gynaecological condition. The Study population were inpatients admitted with a primary obstetric or gynaecological problem who had a concomitant cough and were able to expectorate a sputum sample. Sputum samples from 94 patients were analysed for the presence of Mycobacterium tuberculosis (M.tb) by standard smear microscopy, MGIT culture, MGIT drug-susceptibility testing (DST) and the Xpert(®) MTB/RIF assay. The sensitivity and specificity of the Xpert(®) MTB/RIF assay were evaluated against the culture gold standard. Twenty-six of 94 (27.7%) patients had culture-confirmed pulmonary TB. The Xpert(®) MTB/RIF assay had a sensitivity of 80.8% [95% CI: 60.0-92.7%]) compared against MGIT culture. The Xpert(®) MTB/RIF assay was more sensitive than sputum smear microscopy (21/26 (80.8%) vs. 13/26 (50.0%), P = 0.02) and detected an additional eight culture-confirmed cases. Culture DST analysis identified two monoresistant M.tb strains: one resistant to rifampicin (rifampicin sensitive by the Xpert(®) MTB/RIF assay) and one to ethambutol. HIV infection was linked with a 3-fold increase in risk of TB, accounting for 87.5% (21/24) of TB cases. 50% of cases presented as comorbidities with other communicable diseases (CDs) and non-communicable diseases (NCDs). As an alternative to sputum microscopy, the Xpert(®) MTB/RIF assay provides a sensitive, specific and rapid method for the diagnosis of pulmonary TB in obstetric or gynaecological inpatients. Pulmonary TB is an important cause of concomitant comorbidity to the obstetric or gynaecological condition necessitating admission. TB and HIV comorbidities with other communicable and non-communicable diseases were also common. More proactive screening for TB comorbidity is required in obstetric and gynaecological wards. © 2013 John Wiley & Sons Ltd.

Systematic assessment of blood circulation time of functionalized upconversion nanoparticles in the chick embryo

NASA Astrophysics Data System (ADS)

Nadort, Annemarie; Liang, Liuen; Grebenik, Ekaterina; Guller, Anna; Lu, Yiqing; Qian, Yi; Goldys, Ewa; Zvyagin, Andrei

2015-12-01

Nanoparticle-based delivery of drugs and contrast agents holds great promise in cancer research, because of the increased delivery efficiency compared to `free' drugs and dyes. A versatile platform to investigate nanotechnology is the chick embryo chorioallantoic membrane tumour model, due to its availability (easy, cheap) and accessibility (interventions, imaging). In our group, we developed this model using several tumour cell lines (e.g. breast cancer, colon cancer). In addition, we have synthesized in-house silica coated photoluminescent upconversion nanoparticles with several functional groups (COOH, NH2, PEG). In this work we will present the systematic assessment of their in vivo blood circulation times. To this end, we injected chick embryos grown ex ovo with the functionalized UCNPs and obtained a small amount of blood at several time points after injection to create blood smears The UCNP signal from the blood smears was quantified using a modified inverted microscope imaging set-up. The results of this systematic study are valuable to optimize biochemistry protocols and guide nanomedicine advancement in the versatile chick embryo tumour model.

Factors associated with negative T-SPOT.TB results among smear-negative tuberculosis patients in China.

PubMed

Kang, Wanli; Wu, Meiying; Yang, Kunyun; Ertai, A; Wu, Shucai; Geng, Shujun; Li, Zhihui; Li, Mingwu; Pang, Yu; Tang, Shenjie

2018-03-09

We compared the positive rates of T-SPOT.TB and bacterial culture in the smear-negative PTB, and analyzed the factors affecting the results of negative T-SPOT.TB and bacterial culture. Retrospective evaluation of data from smear-negative PTB patients who underwent T-SPOT.TB and bacterial culture were done. The agreement and concordance were analyzed between T-SPOT.TB and bacterial culture. Multivariable logistic regression analysis was used to explore the factors associated with positive results of T-SPOT.TB and bacterial culture in smear-negative PTB. 858 eligible smear-negative PTB patients were included in the study. The agreement rate was 25.6% (22.7~28.5%) between T-SPOT.TB and bacterial culture in smear- negative PTB patients. The positive rate of T-SPOT.TB was higher than that of bacterial culture in smear-negative PTB patients (p < 0.001). There were nearly no concordance between T-SPOT.TB and bacterial culture (p > 0.05). Using multivariable logistic regression analysis we found that older age ≥ 60 years (OR = 0.469, 95% CI: 0.287-0.768) and decreased albumin (OR = 0.614, 95% CI: 0.380-0.992) were associated with negative diagnostic results of T-SPOT.TB in smear-negative PTB patients. Female (OR = 0.654, 95% CI: 0.431-0.992) were associated with negative diagnostic results of bacteria culture in smear-negative PTB patients. Our results indicated that the older age and decreased albumin were independently associated with negative T-SPOT.TB responses.

The agreement between self-reported cervical smear abnormalities and screening programme records.

PubMed

Canfell, Karen; Beral, Valerie; Green, Jane; Cameron, Rebecca; Baker, Krys; Brown, Anna

2006-01-01

The Million Women Study is a cohort study of women aged 50-64 years in England and Scotland. As a component of the follow-up questionnaire, participants were asked to indicate if they had an abnormal cervical smear in the previous five years. This study compared self-reported cervical abnormalities with screening records obtained from the National Health Service Cervical Screening Programme. For 1944 randomly selected Million Women Study participants in Oxfordshire, screening records were assessed over a six-year period prior to the date of self-reporting. The six-year period was chosen to allow for errors in the recall of timing of abnormal smears. A total of 68 women (3.5%) had a record of at least one equivocal or abnormal smear within the last six years, whereas 49 women (2.5%) self-reported an abnormality. There was a strong trend for an increased probability of self-reporting a history of an abnormal smear as the severity of the recorded abnormality increased (P <0.001). For women with an NHS record of borderline dyskaryosis, mild dyskaryosis, or moderate dyskaryosis/severe dyskaryosis/invasive cancer, the proportions reporting an abnormality were 40%, 58% and 77%, respectively. For women with negative and inadequate smears, the proportion self-reporting an abnormality were 0.6% and 0.7%, respectively. These results indicate that among women whose screening programme records show an abnormal smear, the proportion self-reporting an abnormality increases with the severity of the recorded lesion. Almost all women with a record of negative or inadequate smear(s) correctly interpret the result and do not self-report an abnormality.

Agar Block Smear Preparation: a Novel Method of Slide Preparation for Preservation of Native Fungal Structures for Microscopic Examination and Long-Term Storage▿

PubMed Central

Woo, Patrick C. Y.; Ngan, Antonio H. Y.; Chui, Hon-Kit; Lau, Susanna K. P.; Yuen, Kwok-Yung

2010-01-01

We describe a novel method of fungal slide preparation named “agar block smear preparation.” A total of 510 agar block smears of 25 fungal strains obtained from culture collections, 90 QC fungal strains, and 82 clinical fungal strains from our clinical microbiology laboratory, which included a total of 137 species of yeasts, molds, and thermal dimorphic fungi, were prepared and examined. In contrast to adhesive tape preparation, agar block smears preserved the native fungal structures, such as intact conidiophores of Aspergillus species and arrangements of conidia in Scopulariopsis brevicaulis. Furthermore, agar block smears allowed examination of fungal structures embedded in the agar, such as the ascomata with ascomal hairs in Chaetomium funicola; pycnidium of Phoma glomerata; the intercalary ovoidal chlamydospores arranged in chains of Fusarium dimerum; and the lateral, spherical chlamydospores arranged in pairs of Fusarium solani. After 1 year of storage, morphological integrity was found to have been maintained in 459 (90%) of the 510 agar block smears. After 3 years of storage, morphological integrity was found to have been maintained in 72 (71%) of the 102 smears prepared in 2006. Agar block smear preparation preserves the native fungal structures and allows long-term storage and examination of fungal structures embedded in the agar, hence overcoming the major drawbacks of adhesive tape preparation. The major roles of agar block smear should be diagnosis for difficult cases, accurate identification of fungal species for clinical management of patients and epidemiological studies, and long-term storage for transportation of slides and education purposes. PMID:20660221

Agar block smear preparation: a novel method of slide preparation for preservation of native fungal structures for microscopic examination and long-term storage.

PubMed

Woo, Patrick C Y; Ngan, Antonio H Y; Chui, Hon-Kit; Lau, Susanna K P; Yuen, Kwok-Yung

2010-09-01

We describe a novel method of fungal slide preparation named "agar block smear preparation." A total of 510 agar block smears of 25 fungal strains obtained from culture collections, 90 QC fungal strains, and 82 clinical fungal strains from our clinical microbiology laboratory, which included a total of 137 species of yeasts, molds, and thermal dimorphic fungi, were prepared and examined. In contrast to adhesive tape preparation, agar block smears preserved the native fungal structures, such as intact conidiophores of Aspergillus species and arrangements of conidia in Scopulariopsis brevicaulis. Furthermore, agar block smears allowed examination of fungal structures embedded in the agar, such as the ascomata with ascomal hairs in Chaetomium funicola; pycnidium of Phoma glomerata; the intercalary ovoidal chlamydospores arranged in chains of Fusarium dimerum; and the lateral, spherical chlamydospores arranged in pairs of Fusarium solani. After 1 year of storage, morphological integrity was found to have been maintained in 459 (90%) of the 510 agar block smears. After 3 years of storage, morphological integrity was found to have been maintained in 72 (71%) of the 102 smears prepared in 2006. Agar block smear preparation preserves the native fungal structures and allows long-term storage and examination of fungal structures embedded in the agar, hence overcoming the major drawbacks of adhesive tape preparation. The major roles of agar block smear should be diagnosis for difficult cases, accurate identification of fungal species for clinical management of patients and epidemiological studies, and long-term storage for transportation of slides and education purposes.

Usefulness of interferon-γ release assay for the diagnosis of sputum smear-negative pulmonary and extra-pulmonary TB in Zhejiang Province, China.

PubMed

Ji, Lei; Lou, Yong-Liang; Wu, Zhong-Xiu; Jiang, Jin-Qin; Fan, Xing-Li; Wang, Li-Fang; Liu, Xiao-Xiang; Du, Peng; Yan, Jie; Sun, Ai-Hua

2017-09-01

Quick diagnosis of smear-negative pulmonary tuberculosis (TB) and extra-pulmonary TB are urgently needed in clinical diagnosis. Our research aims to investigate the usefulness of the interferon-γ release assay (IGRA) for the diagnosis of smear-negative pulmonary and extra-pulmonary TB. We performed TB antibody and TB-IGRA tests on 389 pulmonary TB patients (including 120 smear-positive pulmonary TB patients and 269 smear-negative pulmonary TB patients), 113 extra-pulmonary TB patients, 81 patients with other pulmonary diseases and 100 healthy controls. Blood samples for the TB-Ab test and the TB-IGRA were collected, processed, and interpreted according to the manufacturer's protocol. The detection ratio of smear-positive pulmonary TB patients and smear-negative pulmonary TB patients were 90.8% (109 of 120) and 89.6% (241 of 269), respectively. There was no statistically significant difference of its performance between these two sample sets (P > 0.05). The detection ratio of positive TB patients and extra-pulmonary TB patients were 90.0% (350 of 389) and 87.6% (99 of 113), respectively, which was not significantly different (P > 0.05). In this work, the total detection ratio using TB-IGRA was 89.4%, therefore TB-IGRA has diagnostic values in smear-negative pulmonary TB and extra-pulmonary TB diagnosis.

The utilization and barriers of Pap smear among women with visual impairment.

PubMed

Fang, Wen-Hui; Yen, Chia-Feng; Hu, Jung; Lin, Jin-Ding; Loh, Ching-Hui

2016-04-12

Many evidences illustrate that the Pap smear screening successfully reduces if the cervical cancer could be detected and treated sufficiently early. People with disability were higher comorbidity prevalence, and less likely to use preventive health care and health promotion activities. There were also to demonstrate that people with visual impairment has less access to appropriate healthcare services and is less likely to receive screening examinations. In Taiwan, there was no study to explore utilization of Pap smear, associated factors and use barriers about Pap smear screening test among women with visual impairment. The purpose is to explore the utilization and barriers of using Pap smear for women with visual impairment in Taiwan. To identify the barriers of women with visual from process of receiving Pap smear screening test. The cross-sectional study was conducted and the totally 316 participators were selected by stratified proportional and random sampling from 15 to 64 year old women with visual impairment who lived in Taipei County during December 2009 to January 2010. The data was been collected by phone interview and the interviewers were well trained before interview. The mean age was 47.1 years old and the highest percentage of disabled severity was mile (40.2 %). Totally, 66.5 % of participators were ever using Pap smear and 38.9 % used it during pass 1 year. Their first time to accept Pap smear was 38.8 year old. There was near 50 % of them not to be explained by professionals before accepting the Pap smear. For non-using cases, the top two percentage of barriers were "feel still younger" (22.3 %), the second was "there's no sexual experience" (21.4 %). We found the gynecology experiences was key factor for women with visual impairment to use Pap smear, especially the experiences was during 1 year (OR = 4). Associated factors and barriers to receive Pap smear screening test for women with visual impairment can be addressed through interventions aimed at improving on cognitions and attitudes for cervical cancer risk factors. Our study would be as a reference resource for erasing the barriers and inequality among the visually disabled women.

Crystallization and demineralization phenomena in washed-rind cheese.

PubMed

Tansman, Gil F; Kindstedt, Paul S; Hughes, John M

2017-11-01

This report documents an observational study of a high-moisture washed-rind cheese. Three batches of cheese were sampled on a weekly basis for 6 wk and again at wk 10. Center, under-rind, rind, and smear samples were tested for pH, moisture, and selected mineral elements. Powder x-ray diffractometry and petrographic microscopy were applied to identify and image the crystal phases. The pH of the rind increased by over 2 pH units by wk 10. The pH of the under-rind increased but remained below the rind pH, whereas the center pH decreased for most of aging and only began to rise after wk 5. Diffractograms of smear material revealed the presence of 4 crystal phases: brushite, calcite, ikaite, and struvite. The phases nucleated in succession over the course of aging, with calcite and ikaite appearing around the same time. A very small amount of brushite appeared sporadically in center and under-rind samples, but otherwise no other crystallization was observed beneath the rind. Micrographs revealed that crystals in the smear grew to over 250 μm in length by wk 10, and at least 2 different crystal phases, probably ikaite and struvite, could be differentiated by their different optical properties. The surface crystallization was accompanied by a mineral diffusion phenomenon that resulted, on average, in a 217, 95.7, and 149% increase in calcium, phosphorus, and magnesium, respectively, in the rind by wk 10. The diffusion phenomenon caused calcium, phosphorus, and magnesium to decrease, on average, by 55.0, 21.5, and 36.3%, respectively, in the center by wk 10. The present study represents the first observation of crystallization and demineralization phenomena in washed-rind cheese. The Authors. Published by the Federation of Animal Science Societies and Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

Performance of the G4 Xpert® MTB/RIF assay for the detection of Mycobacterium tuberculosis and rifampin resistance: a retrospective case-control study of analytical and clinical samples from high- and low-tuberculosis prevalence settings.

PubMed

Dharan, Nila J; Blakemore, Robert; Sloutsky, Alex; Kaur, Devinder; Alexander, Richard C; Ghajar, Minoo; Musser, Kimberlee A; Escuyer, Vincent E; Rowlinson, Marie-Claire; Crowe, Susanne; Laniado-Laborin, Rafael; Valli, Eloise; Nabeta, Pamela; Johnson, Pamela; Alland, David

2016-12-20

The Xpert ® MTB/RIF (Xpert) assay is a rapid PCR-based assay for the detection of Mycobacterium tuberculosis complex DNA (MTBc) and mutations associated with rifampin resistance (RIF). An updated version introduced in 2011, the G4 Xpert, included modifications to probe B and updated analytic software. An analytical study was performed to assess Xpert detection of mutations associated with rifampin resistance in rifampin-susceptible and -resistant isolates. A clinical study was performed in which specimens from US and non-US persons suspected of tuberculosis (TB) were tested to determine Xpert performance characteristics. All specimens underwent smear microscopy, mycobacterial culture, conventional drug-susceptibility testing and Xpert testing; DNA from isolates with discordant rifampin resistance results was sequenced. Among 191 laboratory-prepared isolates in the analytical study, Xpert sensitivity for detection of rifampin resistance associated mutations was 97.7% and specificity was 90.8%, which increased to 99.0% after DNA sequencing analysis of the discordant samples. Of the 1,096 subjects in the four clinical studies, 49% were from the US. Overall, Xpert detected MTBc in 439 of 468 culture-positive specimens for a sensitivity of 93.8% (95% confidence interval [CI]: 91.2%-95.7%) and did not detect MTBc in 620 of 628 culture-negative specimens for a specificity of 98.7% (95% CI: 97.5%-99.4%). Sensitivity was 99.7% among smear-positive cases, and 76.1% among smear-negative cases. Non-determinate MTBc detection and false-positive RIF resistance results were low (1.2 and 0.9%, respectively). The updated Xpert assay retained the high sensitivity and specificity of the previous assay versions and demonstrated low rates of non-determinate and RIF resistance false positive results.

Prevalence of smear positive pulmonary tuberculosis and associated risk factors among prisoners in Hadiya Zone prison, Southern Ethiopia.

PubMed

Fuge, Terefe G; Ayanto, Samuel Y

2016-04-02

People concentrated in congregated systems such as prisons, are important but often neglected reservoirs for tuberculosis transmission, and threaten those in the outside community. The condition is more serious in Africa particularly in Sub-Saharan Africa (SSA) due to its poor living conditions and ineffective health services. This study was conducted to determine the prevalence of smear positive pulmonary tuberculosis and associated risk factors among prisoners in Hadiya Zone prison. A cross-sectional survey was carried out from May to June 2013 in Hadiya Zone prison. Prison inmates who had history of cough for at least a week were included in the study. Three morning sputum samples were collected from suspected inmates and examined through compound light microscopy. The data obtained was analyzed using statistical software like Epidata and STATA. A total of 164 prisoners were included in the survey using active screening strategy and the point prevalence of smear positive pulmonary tuberculosis (PTB) in the prison was 349.2 per 100,000 populations; about three times higher than its prevalence in the general population. Even though lack of visit from family was the only variable identified as a risk factor for PTB (P = 0.029), almost all of the PTB positive cases were rural residents, farmers, male youngsters and those who shared cell with TB patients and chronically coughing persons as well as those who stayed in a cell that contains >100 inmates. There is high prevalence of TB in Hadiya Zone prison with possible active transmission of TB within the prison. The study also documented a number of factors which may facilitate exposures to TB though most of them are not significantly associated. Therefore, strong cooperation between prison authorities and the national tuberculosis control programmes is urgently required to develop locally appropriate interventions to reduce transmission.

Comparative performance of Thin Layer Agar and Löwenstein-Jensen culture for diagnosis of tuberculosis.

PubMed

Battaglioli, T; Rintiswati, N; Martin, A; Palupi, K R; Bernaerts, G; Dwihardiani, B; Ahmad, R A; Matthys, F; Mahendradhata, Y; Van der Stuyft, P

2013-11-01

Sputum smear microscopy for the diagnosis of tuberculosis (TB) is cheap and simple but its sensitivity is low. Culture on Löwenstein-Jensen (LJ) is more sensitive but it takes a long time to yield results. Thin-Layer Agar (TLA) culture was suggested as an equally sensitive and faster alternative. We evaluated the performance of TLA for diagnosing TB in Jogjakarta, Indonesia. People with suspected TB presenting from July 2010 to July 2011 to two chest clinics of the National TB Control Programme network of Jogjakarta were eligible for inclusion. A sputum sample was sent to the Gadjah Mada University microbiology laboratory for concentration, smearing, Ziehl-Neelsen staining and culture on LJ and TLA. Sensitivity of cultures was evaluated against a composite reference standard (any positive culture). Time to detection of Mycobacteria was recorded. Out of 1414 samples, 164 (12%) were smear positive, 99 (7%) were scanty and 1151 (81%) were negative. On TLA and LJ respectively, 168 (12%) and 149 (11%) samples were positive, 72 (5%) and 32 (2%) were contaminated (κ = 0.64; 95% CI 0.59-0.69, p <0.01). Using the reference standard, 196 (14%) TB cases were identified. The sensitivity of TLA was 0.86 (95% CI 0.80-0.90), significantly higher (p 0.03) than for LJ (0.76; 95% CI 0.69-0.81). The median time to detection in days was significantly shorter (p <0.01) for TLA (12; 95% CI 11-13) than for LJ (44; 95% CI 43-45). TLA is a rapid and sensitive method for the diagnosis of TB. Implementation studies to evaluate the cost-effectiveness and impact of its introduction into programmatic settings are urgently needed. © 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.

Burden of asymptomatic malaria among a tribal population in a forested village of central India: a hidden challenge for malaria control in India.

PubMed

Chourasia, M K; Raghavendra, K; Bhatt, R M; Swain, D K; Valecha, N; Kleinschmidt, I

2017-06-01

Chhattisgarh in India is a malaria-endemic state with seven southern districts that contributes approximately 50-60% of the reported malaria cases in the state every year. The problem is further complicated due to asymptomatic malaria cases which are largely responsible for persistent transmission. This study was undertaken in one of the forested villages of the Keshkal subdistrict in Kondagaon district to ascertain the proportion of the population harbouring subclinical malarial infections. Community-based cross-sectional study. Mass blood surveys were undertaken of the entire population of the village in the post-monsoon seasons of 2013 and 2014. Fingerprick blood smears were prepared from individuals of all ages to detect malaria infections in their blood. Individuals with fever at the time of the survey were tested with rapid diagnostic tests, and parasitaemia in thick blood smears was confirmed by microscopy. Malaria-positive cases were treated with anti-malarials in accordance with the national drug policy. Peripheral blood smears of 134 and 159 individuals, including children, were screened for malaria infection in 2013 and 2014, respectively. Overall, the malaria slide positivity rates were 27.6% and 27.7% in 2013 and 2014, respectively, and the prevalence rates of asymptomatic malaria were 20% and 22.8%. This study showed that, for two consecutive years, the prevalence of asymptomatic malaria infection was significantly higher among children aged ≤14 years (34.4% and 34.1% for 2013 and 2014, respectively) compared with adults (15.2% and 18.2% for 2013 and 2014, respectively; P = 0.023 and 0.04, respectively). The number of asymptomatic malaria cases, especially Plasmodium falciparum, is significant, reinforcing the underlying challenge facing the malaria elimination programme in India. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

[Human resource capacity building on TB laboratory work for TB control program--through the experience of international TB laboratory training course for TB control at the Research Institute of Tuberculosis, JATA, Japan].

PubMed

Fujiki, Akiko; Kato, Seiya

2008-06-01

The international training course on TB laboratory work for national tuberculosis program (NTP) has been conducted at the Research Institute of Tuberculosis since 1975 funded by Japan International Cooperation Agency in collaboration with WHO Western Pacific Regional Office. The aim of the course is to train key personnel in TB laboratory field for NTP in resource-limited countries. The course has trained 265 national key personnel in TB laboratory service from 57 resource-limited countries in the last 33 years. The number of participants trained may sound too small in the fight against the large TB problem in resource-limited countries. However, every participant is playing an important role as a core and catalyst for the TB control program in his/her own country when they were back home. The curriculum is composed of technical aspects on TB examination, mainly sputum microscopy in addition since microscopy service is provided at many centers that are deployed in a widely spread area, the managerial aspect of maintaining quality TB laboratory work at the field laboratory is another component of the curriculum. Effective teaching methods using materials such as artificial sputum, which is useful for panel slide preparation, and technical manuals with illustrations and pictures of training procedure have been developed through the experience of the course. These manuals are highly appreciated and widely used by the front line TB workers. The course has also contributed to the expansion of EQA (External Quality Assessment) system on AFB microscopy for the improvement of the quality of TB laboratory service of NTP. The course is well-known for not only having a long history, but also for its unique learning method emphasizing "Participatory Training", particularly for practicum sessions to master the skills on AFB microscopy. The method in learning AFB microscopy, which was developed by the course, was published as a training manual by IUATLD, RIT and USAID. As it is mentioned, the course has been contributing to human resource capacity building including management of laboratory service to improve NTP in the resource-limited countries. Currently, expansion of technology transfer on culture examination for drug susceptibility test has been attempted to the resource-limited countries due to the occurrence of MDR-TB (Multi drug-resistant tuberculosis) and XDR-TB (Extensively drug-resistant tuberculosis) cases. However, since sputum smear examination is most effective method of detection of infectious TB, the writers believe it is still a core component of TB control, unless a new diagnostic tool that is practicable and effective in the resource-limited countries is developed. Therefore the course will keep focused on the smear examination as the basic curriculum. The course is highly appreciated by international experts and it is our responsibility to answer the expectation from them.

A novel scheme for abnormal cell detection in Pap smear images

NASA Astrophysics Data System (ADS)

Zhao, Tong; Wachman, Elliot S.; Farkas, Daniel L.

2004-07-01

Finding malignant cells in Pap smear images is a "needle in a haystack"-type problem, tedious, labor-intensive and error-prone. It is therefore desirable to have an automatic screening tool in order that human experts can concentrate on the evaluation of the more difficult cases. Most research on automatic cervical screening tries to extract morphometric and texture features at the cell level, in accordance with the NIH "The Bethesda System" rules. Due to variances in image quality and features, such as brightness, magnification and focus, morphometric and texture analysis is insufficient to provide robust cervical cancer detection. Using a microscopic spectral imaging system, we have produced a set of multispectral Pap smear images with wavelengths from 400 nm to 690 nm, containing both spectral signatures and spatial attributes. We describe a novel scheme that combines spatial information (including texture and morphometric features) with spectral information to significantly improve abnormal cell detection. Three kinds of wavelet features, orthogonal, bi-orthogonal and non-orthogonal, are carefully chosen to optimize recognition performance. Multispectral feature sets are then extracted in the wavelet domain. Using a Back-Propagation Neural Network classifier that greatly decreases the influence of spurious events, we obtain a classification error rate of 5%. Cell morphometric features, such as area and shape, are then used to eliminate most remaining small artifacts. We report initial results from 149 cells from 40 separate image sets, in which only one abnormal cell was missed (TPR = 97.6%) and one normal cell was falsely classified as cancerous (FPR = 1%).

A Study of Pap Smear in HIV-Positive Females.

PubMed

Madan, Apeksha; Patil, Sunita; Nakate, Leena

2016-12-01

HIV-positive females are more likely to have abnormal Pap smears than HIV-negative women. These abnormal Pap smears are usually associated with low CD4 cell counts and human papilloma virus infection. This was a prospective hospital-based study from April 2013 till March 2014. A total of 250 (both symptomatic and asymptomatic) HIV-positive females were examined in Gynaecology OPD at R.C.S.M. G.M.C and C.P.R. Hospital, Kolhapur, and their cervical smears were taken. They were categorized as per modified Bethesda system 2001. The findings in HIV-positive women were correlated with risk factors (age, disease duration, CD4 count and ART use). To study the spectrum of cytological abnormalities on Pap smear in HIV-positive females and classify precancerous and cancerous lesions in HIV-positive females according to Bethesda system 2001 and to be familiar with terminology and morphological criteria of Bethesda system 2001. To study the association of Pap smears abnormalities among HIV-positive women with their immune status (CD4 count). NILM is the commonest finding (83.2 %) which is subdivided into non-inflammatory, non-specific and specific inflammatory and atrophic smears. Candida vaginitis was the commonest cause of specific inflammatory condition accounted for (2.52 %) of all inflammatory smears. The percentage of squamous cell abnormalities was 12 %: ASCUS + ASC-H-6.22 %, LSIL-2.10 %, HSIL-3.4 % and SCC-0.8 %. The highest incidence of intraepithelial lesions in HIV-positive females was in the age group 31-40 years. There is no association of Pap smear abnormalities among HIV-positive women with their immune status (CD4 count) and duration of ART. Periodic, regular gynaecologic and Pap smear examination would help in early detection of intraepithelial lesions and their treatment so as to prevent invasive malignancy and mortality.

GenoType® Mtbdrsl assay for resistance to second-line anti-tuberculosis drugs

PubMed Central

Theron, Grant; Peter, Jonny; Richardson, Marty; Warren, Rob; Dheda, Keertan; Steingart, Karen R

2016-01-01

Background Genotype® MTBDRsl (MTBDRsl) is a rapid DNA-based test for detecting specific mutations associated with resistance to fluoroquinolones and second-line injectable drugs (SLIDs) in Mycobacterium tuberculosis complex. MTBDRsl version 2.0 (released in 2015) identifies the mutations detected by version 1.0, as well as additional mutations. The test may be performed on a culture isolate or a patient specimen, which eliminates delays associated with culture. Version 1.0 requires a smear-positive specimen, while version 2.0 may use a smear-positive or -negative specimen. We performed this updated review as part of a World Health Organization process to develop updated guidelines for using MTBDRsl. Objectives To assess and compare the diagnostic accuracy of MTBDRsl for: 1. fluoroquinolone resistance, 2. SLID resistance, and 3. extensively drug-resistant tuberculosis, indirectly on a M. tuberculosis isolate grown from culture or directly on a patient specimen. Participants were people with rifampicin-resistant or multidrug-resistant tuberculosis. The role of MTBDRsl would be as the initial test, replacing culture-based drug susceptibility testing (DST), for detecting second-line drug resistance. Search methods We searched the following databases without language restrictions up to 21 September 2015: the Cochrane Infectious Diseases Group Specialized Register; MEDLINE; Embase OVID; Science Citation Index Expanded, Conference Proceedings Citation Index-Science, and BIOSIS Previews (all three from Web of Science); LILACS; and SCOPUS; registers for ongoing trials; and ProQuest Dissertations & Theses A&I. We reviewed references from included studies and contacted specialists in the field. Selection criteria We included cross-sectional and case-control studies that determined MTBDRsl accuracy against a defined reference standard (culture-based DST, genetic sequencing, or both). Data collection and analysis Two review authors independently extracted data and assessed quality using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) tool. We synthesized data for versions 1.0 and 2.0 separately. We estimated MTBDRsl sensitivity and specificity for fluoroquinolone resistance, SLID resistance, and extensively drug-resistant tuberculosis when the test was performed indirectly or directly (smear-positive specimen for version 1.0, smear-positive or -negative specimen for version 2.0). We explored the influence on accuracy estimates of individual drugs within a drug class and of different reference standards. We performed most analyses using a bivariate random-effects model with culture-based DST as reference standard. Main results We included 27 studies. Twenty-six studies evaluated version 1.0, and one study version 2.0. Of 26 studies stating specimen country origin, 15 studies (58%) evaluated patients from low- or middle-income countries. Overall, we considered the studies to be of high methodological quality. However, only three studies (11%) had low risk of bias for the reference standard; these studies used World Health Organization (WHO)-recommended critical concentrations for all drugs in the culture-based DST reference standard. MTBDRsl version 1.0 Fluoroquinolone resistance: indirect testing, MTBDRsl pooled sensitivity and specificity (95% confidence interval (CI)) were 85.6% (79.2% to 90.4%) and 98.5% (95.7% to 99.5%), (19 studies, 2223 participants); direct testing (smear-positive specimen), pooled sensitivity and specificity were 86.2% (74.6% to 93.0%) and 98.6% (96.9% to 99.4%), (nine studies, 1771 participants, moderate quality evidence). SLID resistance: indirect testing, MTBDRsl pooled sensitivity and specificity were 76.5% (63.3% to 86.0%) and 99.1% (97.3% to 99.7%), (16 studies, 1921 participants); direct testing (smear-positive specimen), pooled sensitivity and specificity were 87.0% (38.1% to 98.6%) and 99.5% (93.6% to 100.0%), (eight studies, 1639 participants, low quality evidence). Extensively drug-resistant tuberculosis: indirect testing, MTBDRsl pooled sensitivity and specificity were 70.9% (42.9% to 88.8%) and 98.8% (96.1% to 99.6%), (eight studies, 880 participants); direct testing (smear-positive specimen), pooled sensitivity and specificity were 69.4% (38.8% to 89.0%) and 99.4% (95.0% to 99.3%), (six studies, 1420 participants, low quality evidence). Similar to the original Cochrane review, we found no evidence of a significant difference in MTBDRsl version 1.0 accuracy between indirect and direct testing for fluoroquinolone resistance, SLID resistance, and extensively drug-resistant tuberculosis. MTBDRsl version 2.0 Fluoroquinolone resistance: direct testing, MTBDRsl sensitivity and specificity were 97% (83% to 100%) and 98% (93% to 100%), smear-positive specimen; 80% (28% to 99%) and 100% (40% to 100%), smear-negative specimen. SLID resistance: direct testing, MTBDRsl sensitivity and specificity were 89% (72% to 98%) and 90% (84% to 95%), smear-positive specimen; 80% (28% to 99%) and 100% (40% to 100%), smear-negative specimen. Extensively drug-resistant tuberculosis: direct testing, MTBDRsl sensitivity and specificity were 79% (49% to 95%) and 97% (93% to 99%), smear-positive specimen; 50% (1% to 99%) and 100% (59% to 100%), smear-negative specimen. We had insufficient data to estimate summary sensitivity and specificity of version 2.0 (smear-positive and -negative specimens) or to compare accuracy of the two versions. A limitation was that most included studies did not consistently use the World Health Organization (WHO)-recommended concentrations for drugs in the culture-based DST reference standard. Authors' conclusions In people with rifampicin-resistant or multidrug-resistant tuberculosis, MTBDRsl performed on a culture isolate or smear-positive specimen may be useful in detecting second-line drug resistance. MTBDRsl (smear-positive specimen) correctly classified around six in seven people as having fluoroquinolone or SLID resistance, although the sensitivity estimates for SLID resistance varied. The test rarely gave a positive result for people without drug resistance. However, when second-line drug resistance is not detected (MTBDRsl result is negative), conventional DST can still be used to evaluate patients for resistance to the fluoroquinolones or SLIDs. We recommend that future work evaluate MTBDRsl version 2.0, in particular on smear-negative specimens and in different settings to account for different resistance-causing mutations that may vary by strain. Researchers should also consider incorporating WHO-recommended critical concentrations into their culture-based reference standards. PLAIN LANGUAGE SUMMARY The rapid test GenoType® MTBDRsl for testing resistance to second-line TB drugs Background Different drugs are available to treat tuberculosis (TB), but resistance to these drugs is a growing problem. People with drug-resistant TB require second-line TB drugs that, compared with first-line TB drugs, must be taken for longer and may be associated with more harms. Detecting TB drug resistance quickly is important for improving health, reducing deaths, and decreasing the spread of drug-resistant TB. Definitions Multidrug-resistant TB (MDR-TB) is caused by TB bacteria that are resistant to at least isoniazid and rifampicin, the two most potent TB drugs. Extensively drug-resistant TB (XDR-TB) is a type of MDR-TB that is resistant to nearly all TB drugs. What test is evaluated by this review? GenoType® MTBDRsl (MTBDRsl) is a rapid test for detecting resistance to second-line TB drugs. In people with MDR-TB, MTBDRsl is used to detect additional drug resistance. The test may be performed on TB bacteria grown in culture from a patient specimen (indirect testing) or on a patient specimen (direct testing), which eliminates delays associated with culture. MTBDRsl version 1.0 requires a specimen to be smear-positive by microscopy, while version 2.0 (released in 2015) may use a smear-positive or -negative specimen. What are the aims of the review? We wanted to find out how accurate MTBDRsl is for detecting drug resistance; to compare indirect and direct testing; and to compare the two test versions. How up-to-date is the review? We searched for and used studies that had been published up to 21 September 2015. What are the main results of the review? We found 27 studies; 26 studies evaluated MTBDRsl version 1.0 and one study evaluated version 2.0. Fluoroquinolone drugs MTBDRsl version 1.0 (smear-positive specimen) detected 86% of people with fluoroquinolone resistance and rarely gave a positive result for people without resistance (GRADE, moderate quality evidence). Second-line injectable drugs MTBDRsl version 1.0 (smear-positive specimen) detected 87% of people with second-line injectable drug resistance and rarely gave a positive result for people without resistance (GRADE, low quality evidence). XDR-TB MTBDRsl version 1.0 (smear-positive specimen) detected 69% of people with XDR-TB and rarely gave a positive result for people without resistance (GRADE, low quality evidence). For MTBDRsl version 1.0, we found similar results for indirect and direct testing (smear-positive specimen). As we identified only one study evaluating MTBDRsl version 2.0, we could not be sure of the diagnostic accuracy of version 2.0. Also, we could not compare accuracy of the two versions. What is the methodological quality of the evidence? We used the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) tool to assess study quality. Overall, we considered the included studies to be of high quality; however, we had concerns about how the reference standard (the benchmark against which MTBDRsl was measured) was applied. What are the authors' conclusions? MTBDRsl (smear-positive specimen) identified most of the patients with second-line drug resistance. When the test reports a negative result, conventional testing for drug resistance can still be used. PMID:27605387

Pilot Study of an Open-source Image Analysis Software for Automated Screening of Conventional Cervical Smears.

PubMed

Sanyal, Parikshit; Ganguli, Prosenjit; Barui, Sanghita; Deb, Prabal

2018-01-01

The Pap stained cervical smear is a screening tool for cervical cancer. Commercial systems are used for automated screening of liquid based cervical smears. However, there is no image analysis software used for conventional cervical smears. The aim of this study was to develop and test the diagnostic accuracy of a software for analysis of conventional smears. The software was developed using Python programming language and open source libraries. It was standardized with images from Bethesda Interobserver Reproducibility Project. One hundred and thirty images from smears which were reported Negative for Intraepithelial Lesion or Malignancy (NILM), and 45 images where some abnormality has been reported, were collected from the archives of the hospital. The software was then tested on the images. The software was able to segregate images based on overall nuclear: cytoplasmic ratio, coefficient of variation (CV) in nuclear size, nuclear membrane irregularity, and clustering. 68.88% of abnormal images were flagged by the software, as well as 19.23% of NILM images. The major difficulties faced were segmentation of overlapping cell clusters and separation of neutrophils. The software shows potential as a screening tool for conventional cervical smears; however, further refinement in technique is required.

Smearing the opposition: implicit and explicit stigmatization of the 2008 U.S. Presidential candidates and the current U.S. President.

PubMed

Kosloff, Spee; Greenberg, Jeff; Schmader, Toni; Dechesne, Mark; Weise, David

2010-08-01

Four studies investigated whether political allegiance and salience of outgroup membership contribute to the phenomenon of acceptance of false, stigmatizing information (smears) about political candidates. Studies 1-3 were conducted in the month prior to the 2008 U.S. Presidential election and together demonstrated that pre-standing opposition to John McCain or Barack Obama, as well as the situational salience of differentiating social categories (i.e., for Obama, race; for McCain, age), contributed to the implicit activation and explicit endorsement of smearing labels (i.e., Obama is Muslim; McCain is senile). The influence of salient differentiating categories on smear acceptance was particularly pronounced among politically undecided individuals. Study 4 clarified that social category differences heighten smear acceptance, even if the salient category is semantically unrelated to the smearing label, showing that, approximately 1 year after the election, the salience of race amplified belief that Obama is a socialist among undecided people and McCain supporters. Taken together, these findings suggest that, at both implicit and explicit cognitive levels, social category differences and political allegiance contribute to acceptance of smears against political candidates. 2010 APA, all rights reserved

Acculturation and Pap smear screening practices among college-aged Vietnamese women in the United States.

PubMed

Yi, J K

1998-10-01

The purposes of this study were both to gather information on the prevalence of Pap smear screening among college-aged Vietnamese women and to identify predictive factors that influence Vietnamese women to engage in early cervical cancer detection behavior. This study also examines the role of acculturation in explaining cervical cancer screening behavior. Data were collected through mail and telephone surveys. Of those respondents (n = 207), approximately one-third (36.8%) reported having had a Pap smear. More than one-third (39.3%) of sexually active respondents reported never having had a Pap smear. Acculturated women were more likely to be sexually active and to be obtaining regular Pap smears. In the logistic regression model, marital status, sexual activity, and a belief that only married women should have a Pap smear were important predictors of prior reported Pap smear experience. These study findings suggest that less-acculturated women appear to have the belief that cervical cancer screening is more important for married women than for unmarried women, regardless of their sexual activity. The existence of such misconceptions underscores the importance of offering culturally relevant cancer education that addresses related misconceptions.

One-step purification of Enterocytozoon bieneusi spores from human stools by immunoaffinity expanded-bed adsorption.

PubMed

Accoceberry, I; Thellier, M; Datry, A; Desportes-Livage, I; Biligui, S; Danis, M; Santarelli, X

2001-05-01

An original, reliable, and reproducible method for the purification of Enterocytozoon bieneusi spores from human stools is described. We recently reported the production of a species-specific monoclonal antibody (MAb) 6E52D9 immunoglobulin G2a (IgG2a) raised against the exospore of E. bieneusi spore walls. The MAb was used as a ligand to develop an immunoaffinity matrix. The mouse IgG2a MAb was bound directly to a Streamline rProtein A adsorbent, used for expanded-bed adsorption of immunoglobulins, for optimal spatial orientation of the antibody and maximum binding efficiency of the antigen. The complex was then cross-linked covalently using dimethyl pimelimidate dihydrochloride. After incubation of the immunoaffinity matrix with filtered stool samples containing numerous E. bieneusi spores and before elution with 6 M guanidine HCl, the expansion of the adsorbent bed eliminated all the fecal contaminants. The presence of spores in the elution fractions was determined by an indirect immunofluorescence antibody test (IFAT). E. bieneusi spores were found in the elution fraction in all four experiments and were still highly antigenic as indicated by IFAT. Smears examined by light microscopy contained very clean spores with no fecal debris or background bacterial and fungal contaminants. However, spore recovery rates were relatively low: an average of 10(7) spores were purified per run. This technique for isolating E. bieneusi spores directly from human stool samples with a high degree of purity opens up new approaches for studying this parasite.

Innovatively addressing the challenge of maintaining binocular microscopes under Tuberculosis Programme in India - Is this feasible?

PubMed

Chadha, Sarabjit; Nagaraja, Sharath Burugina; Prasad, Banaru Muralidhara; Kelamane, Santosha; Satyanarayana, Srinath; Sachdeva, Kuldeep Singh

2016-01-01

India is a high TB burden country. The preferred first line diagnostic tool under National TB Programme is sputum smear microscopy through binocular microscopes (BMs) from 13,000 designated microscopy centres across the country. The programme had devised innovative strategy for maintenance of BMs. A study was conducted to look into the operational feasibility of an external agency to provide maintenance services for BMs engaged through newer strategy. A cross-sectional study was conducted in the states of Uttar Pradesh, Bihar and Rajasthan during 2010-2013. A total of 9314 BMs were serviced during the period 2011-2013, of which 1104 (11.8%) had major repairs, 2204 (23.6%) had minor repairs, 1054 (11.3%) were provided emergency breakdown services and 223 (2.4%) were condemned. The bold initiative and newer strategy of the programme to engage agencies for BMs maintenance services is worthwhile and should be continued and could be considered for replication across the country. Copyright © 2016 Tuberculosis Association of India. Published by Elsevier B.V. All rights reserved.

First report of Cystoisospora belli parasitemia in a patient with acquired immunodeficiency syndrome.

PubMed

Velásquez, Jorge Néstor; di Risio, Cecilia Alicia; Etchart, Cristina Beatriz; Chertcoff, Agustín Víctor; Nigro, Mónica Gabriela; Pantano, María Laura; Ledesma, Bibiana Alba; Vittar, Natalia; Carnevale, Silvana

2016-01-01

Cystoisospora belli in patients with the acquired immunodeficiency syndrome (AIDS) has been described as cause of chronic diarrhea and disseminated cystoisosporosis. Diagnosis of intestinal cystoisosporosis can be achieved at the tissue level in the villus epithelium of the small bowel. Disseminated cystoisosporosis is diagnosed by microscopy identification of unizoite tissue cysts in the lamina propria of the intestine. We report a case of disseminated cystoisosporosis in a human immunodeficiency virus (HIV)-infected patient with detection of parasitemia. We studied a 39-year old patient with AIDS and chronic diarrhea by analysis of stool and duodenal biopsy samples. Blood samples were also collected and examined by light microscopy and molecular techniques for C. belli DNA detection. The unizoite tissue cyst stages were present in the lamina propria, with unsporulated oocysts in feces. Zoites were present in blood smears and DNA of C. belli was detected in blood samples. Our study identified a new stage in the life cycle of C. belli. Detection of parasitemia is a novel and noninvasive tool for diagnosis of disseminated cystoisosporosis.

Plasmodium ovale infection in Malaysia: first imported case.

PubMed

Lim, Yvonne A L; Mahmud, Rohela; Chew, Ching Hoong; T, Thiruventhiran; Chua, Kek Heng

2010-10-08

Plasmodium ovale infection is rarely reported in Malaysia. This is the first imported case of P. ovale infection in Malaysia which was initially misdiagnosed as Plasmodium vivax. Peripheral blood sample was first examined by Giemsa-stained microscopy examination and further confirmed using a patented in-house multiplex PCR followed by sequencing. Initial results from peripheral blood smear examination diagnosed P. vivax infection. However further analysis using a patented in-house multiplex PCR followed by sequencing confirmed the presence of P. ovale. Given that Anopheles maculatus and Anopheles dirus, vectors of P. ovale are found in Malaysia, this finding has significant implication on Malaysia's public health sector. The current finding should serve as an alert to epidemiologists, clinicians and laboratory technicians in the possibility of finding P. ovale in Malaysia. P. ovale should be considered in the differential diagnosis of imported malaria cases in Malaysia due to the exponential increase in the number of visitors from P. ovale endemic regions and the long latent period of P. ovale. It is also timely that conventional diagnosis of malaria via microscopy should be coupled with more advanced molecular tools for effective diagnosis.

Smear correction of highly variable, frame-transfer CCD images with application to polarimetry.

PubMed

Iglesias, Francisco A; Feller, Alex; Nagaraju, Krishnappa

2015-07-01

Image smear, produced by the shutterless operation of frame-transfer CCD detectors, can be detrimental for many imaging applications. Existing algorithms used to numerically remove smear do not contemplate cases where intensity levels change considerably between consecutive frame exposures. In this report, we reformulate the smearing model to include specific variations of the sensor illumination. The corresponding desmearing expression and its noise properties are also presented and demonstrated in the context of fast imaging polarimetry.

Cost Analysis of Tuberculosis Diagnosis in Cambodia with and without Xpert® MTB/RIF for People Living with HIV/AIDS and People with Presumptive Multidrug-resistant Tuberculosis.

PubMed

Pallas, Sarah Wood; Courey, Marissa; Hy, Chhaily; Killam, Wm Perry; Warren, Dora; Moore, Brittany

2018-06-04

The Xpert ® MTB/RIF (Xpert) test has been shown to be effective and cost-effective for diagnosing tuberculosis (TB) under conditions with high HIV prevalence and HIV-TB co-infection but less is known about Xpert's cost in low HIV prevalence settings. Cambodia, a country with low HIV prevalence (0.7%), high TB burden, and low multidrug-resistant (MDR) TB burden (1.4% of new TB cases, 11% of retreatment cases) introduced Xpert into its TB diagnostic algorithms for people living with HIV (PLHIV) and people with presumptive MDR TB in 2012. The study objective was to estimate these algorithms' costs pre- and post-Xpert introduction in four provinces of Cambodia. Using a retrospective, ingredients-based microcosting approach, primary cost data on personnel, equipment, maintenance, supplies, and specimen transport were collected at four sites through observation, records review, and key informant consultations. Across the sample facilities, the cost per Xpert test was US$33.88-US$37.11, clinical exam cost US$1.22-US$1.84, chest X-ray cost US$2.02-US$2.14, fluorescent microscopy (FM) smear cost US$1.56-US$1.93, Ziehl-Neelsen (ZN) smear cost US$1.26, liquid culture test cost US$11.63-US$22.83, follow-on work-up for positive culture results and Mycobacterium tuberculosis complex (MTB) identification cost US$11.50-US$14.72, and drug susceptibility testing (DST) cost US$44.26. Specimen transport added US$1.39-US$5.21 per sample. Assuming clinician adherence to the algorithms and perfect test accuracy, the normative cost per patient correctly diagnosed under the post-Xpert algorithms would be US$25-US$29 more per PLHIV and US$34-US$37 more per person with presumptive MDR TB (US$41 more per PLHIV when accounting for variable test sensitivity and specificity). Xpert test unit costs could be reduced through lower cartridge prices, longer usable life of GeneXpert ® (Cepheid, USA) instruments, and increased test volumes; however, epidemiological and test eligibility conditions in Cambodia limit the number of specimens received at laboratories, leading to sub-optimal utilization of current instruments. Improvements to patient referral and specimen transport could increase test volumes and reduce Xpert test unit costs in this setting.

Eosinophilic leukemia in three African pygmy hedgehogs ( Atelerix albiventris) and validation of Luna stain.

PubMed

Martínez-Jiménez, David; Garner, Bridget; Coutermarsh-Ott, Sheryl; Burrell, Caitlin; Clark, Sabrina; Nabity, Mary; Díaz-Delgado, Josué; Rodrigues-Hoffmann, Aline; Zaks, Karen; Proença, Laila; Divers, Stephen; Saba, Corey; Cazzini, Paola

2017-03-01

Neoplasia is usually encountered in the African pygmy hedgehog at a mean age of 3.5 y, and malignancy is common. Myelogenous leukemias are rarely reported in hedgehogs. We describe 3 cases of eosinophilic leukemia in adult, middle-aged (mean age: 2.3 y) hedgehogs, for which prognosis appears grave. In 1 case, attempted treatment was unsuccessful, and in all 3 cases, the disease course was rapid and all died soon after diagnosis. Blood smear evaluation, along with complete blood count, was critical in making the diagnosis in all cases. Luna stain was validated and used to better visualize eosinophils in cytologic and histologic sections. Electron microscopy confirmed the presence of specific granules in hedgehog eosinophils.

Accuracy of next-generation sequencing for the identification of clinically relevant variants in cytology smears in lung adenocarcinoma.

PubMed

Baum, Jordan E; Zhang, Pan; Hoda, Rana S; Geraghty, Brian; Rennert, Hanna; Narula, Navneet; Fernandes, Helen D

2017-06-01

Minimally invasive diagnostic procedures such as needle-core biopsy and fine-needle aspiration provide adequate material for molecular analyses. Advances in precision oncology are trending toward the interrogation of limited amounts of genomic material to guide clinical and therapeutic decisions. The aim of this study was to investigate the minimum cellularity needed on cytologic smears for the identification of clinically relevant variants with next-generation sequencing (NGS). Thirty cases of cytologically diagnosed, resection-proven primary lung adenocarcinoma were identified. Nineteen of the 30 cases were known to harbor actionable variants. One Diff-Quik (DQ)-stained slide and 1 Papanicolaou (Pap)-stained slide were selected from each case. Cases were categorized as containing fewer than 100 tumor cells, 100 to 500 tumor cells, or more than 500 tumor cells. NGS was performed on the Ion Torrent platform. NGS was successfully performed on all cell blocks and on 90% of the smears. Paired DQ and Pap smears showed similar cellularity, and cases that differed in cellularity were within 1 category of each other. The cases with more than 100 tumor cells had a 93% success rate; this was significantly different from the situation for cases with fewer than 100 tumor cells, which were successfully sequenced only 67% of the time. Overall, NGS was able to provide clinically relevant information for 83% of DQ smears and for 90% of Pap smears tested. The data show a significantly higher likelihood of successful NGS with cytologic smears with more than 100 tumor cells. There was a trend for a higher NGS success rate with Pap smears versus DQ smears. Cancer Cytopathol 2017;125:398-406. © 2017 American Cancer Society. © 2017 American Cancer Society.

Prevalence of tuberculous infection and incidence of tuberculosis; a re-assessment of the Styblo rule

PubMed Central

van der Werf, MJ; Borgdorff, MW

2008-01-01

Abstract Objective To evaluate the validity of the fixed mathematical relationship between the annual risk of tuberculous infection (ARTI), the prevalence of smear-positive tuberculosis (TB) and the incidence of smear-positive TB specified as the Styblo rule, which TB control programmes use to estimate the incidence of TB disease at a population level and the case detection rate. Methods Population-based tuberculin surveys and surveys on prevalence of smear-positive TB since 1975 were identified through a literature search. For these surveys, the ratio between the number of tuberculous infections (based on ARTI estimates) and the number of smear-positive TB cases was calculated and compared to the ratio of 8 to 12 tuberculous infections per prevalent smear-positive TB case as part of the Styblo rule. Findings Three countries had national population-based data on both ARTI and prevalence of smear-positive TB for more than one point in time. In China the ratio ranged from 3.4 to 5.8, in the Philippines from 2.6 to 4.4, and in the Republic of Korea, from 3.2 to 4.7. All ratios were markedly lower than the ratio that is part of the Styblo rule. Conclusion According to recent country data, there are typically fewer than 8 to 12 tuberculous infections per prevalent smear-positive TB case, and it remains unclear whether this ratio varies significantly among countries. The decrease in the ratio compared to the Styblo rule probably relates to improvements in the prompt treatment of TB disease (by national TB programmes). A change in the number of tuberculous infections per prevalent smear-positive TB case in population-based surveys makes the assumed fixed mathematical relationship between ARTI and incidence of smear-positive TB no longer valid. PMID:18235886

A comparison of digitized frozen section and smear preparations for intraoperative neurotelepathology.

PubMed

Gould, Peter V; Saikali, Stephan

2012-01-01

Intraoperative consultations in neuropathology are often assessed by smear preparations rather than by frozen sections. Both techniques are standard practice for light microscopic examination on site, but there is little data comparing these techniques in a telepathology setting. Thirty cases of brain tumours submitted for intraoperative consultation at our institution between July and December 2010 were identified in which both frozen section and tissue smear preparations were available for digitization at 20× magnification. Slides were digitized using a Hamamatsu Nanozoomer 2.0 HT whole slide scanner, and resulting digital images were visualized at 1680 × 1050 pixel resolution with NDP. view software. The original intraoperative diagnosis was concordant with the sign out diagnosis in 29/30 cases; one tumeur was initially interpreted as a high grade glioma but proved to be a lymphoma at sign out. Digitized frozen section slides were sufficient for diagnosis at 10× magnification in 27/30 cases. Digitized tissue smears were sufficient for diagnosis at 10× magnification in 28/30 cases. In two cases tumour was present on the tissue smear but not the frozen section (one case of recurrent astrocytoma, one case of meningeal carcinomatosis). In one case of lymphoma, tumour was present on frozen section only. These discrepancies were attributed to tissue sampling rather than image quality. Examination of digitized slides at higher magnfication (20×) permitted confirmation of mitoses and Rosenthal fibers on tissue smear preparations, but did not change the primary diagnosis. Intra-slide variations in tissue thickness on smear preparations led to variable loss of focus in digitized images, but did not affect image quality in thinner areas of the smear or impede diagnosis. Digitized tissue smears are suitable for intraoperative neurotelepathology and provide comparable information to digitized frozen sections at medium power magnification.

Confusion about Pap smears: lack of knowledge among high-risk women.

PubMed

Daley, Ellen; Perrin, Kay; Vamos, Cheryl; Hernandez, Natalie; Anstey, Erica; Baker, Elizabeth; Kolar, Stephanie; Ebbert, Judith

2013-01-01

The impact of the Papanicolaou (Pap) smear on the prevention of cervical cancer is one of the greatest public health success stories. However, it is not clear if women understand the purpose of the Pap smear despite recent advancements and national attention over cervical cancer prevention. The purpose of this study was to examine Pap smear knowledge among three high-risk populations at different points in time. Women from three separate human papillomavirus (HPV) psychosocial studies completed surveys assessing Pap smear knowledge: (1) HPV-positive women (prevaccine population in 2005-2006, n=154, mean age 23.5), (2) college women (postvaccine population in 2008, n=276, mean age 18.9), and (3) minority college women (postvaccine population in 2011, n=711, mean age 23.3). Frequencies and logistic regression were employed to examine associations between demographic factors and accurate knowledge of Pap smear testing within each study. Approximately one quarter of participants across all three samples did not know that the Pap smear is a test for cervical cancer. Participants also incorrectly believed that the Pap smear tests for HPV (82%-91%), vaginal infections (76%-92%), yeast infections (65%-86%), gonorrhea (55%-81%), herpes (53%-80%), HIV/AIDS (22%-59%), and pregnancy (17%-38%). Among all three studies, older age was the only factor significant with higher Pap knowledge. Higher HPV knowledge scores were significantly associated with higher Pap knowledge in studies 2 and 3 only. Knowledge about the purpose of the Pap smear remains low. Findings underscore the significant need for clear and consistent messages among high-risk women regarding the prevention of cervical cancer and other reproductive health conditions.

An evaluation of smear layer with various desensitizing agents after tooth preparation.

PubMed

Zaimoglu, A; Aydin, A K

1992-09-01

According to hydrodynamics, any agent blocking the dentinal tubules reduces the flow of fluids and diminishes hypersensitivity. The properties of the desensitizing agents that sponsor tubular occlusion and the barrier efficiency resulting from the interaction of the smear layer with test materials were examined with the scanning electron microscope and energy-dispersive x-ray microanalysis. Selected dentinal desensitizing was accomplished with burnishing procedures, cavity varnish, calcium hydroxide, and topical fluoride. Subjective evaluations were also recorded clinically after tooth preparation. This investigation indicated that the smear layer did not protect against zinc phosphate cement, and that cavity varnish prevented the formation of the smear plugs. The smear layer and plugs were basically composed of calcium and phosphorus, the major ingredients of dentin.

Pap Smear Test Prevalence within Three Years (Age 18+) - Small Area Estimates

Cancer.gov

For Pap smear test, a woman 18 years of age or older must have reported having at least one Pap smear test in her life. Furthermore, she should have had one within the last three years by the time of interview.

Actinomyces-like organisms in the cervical smears of IUCD users.

PubMed

Farida Jamal; Mohd Salleh Mohd Yasin; Zooraidah Zainal; Hamid Arshat

1983-01-01

Pelvic inflammatory disease (PID) is a well-documented complication of IUD use. The presence of tails on the IUDs appears to facilitate ascent of bacteria from the vagina into the uterus. In this study, cervical smears of 128 IUD users were stained by Gram's stain and screened for the presence of Actinomyces-like organisms. 5.5% of the smears were positive for these organisms. Pus was seen in 20.1% of the smears, and yeast cells were seen in 10.2%. The average duration of IUD insertion was 8 months. Smears of 103 controls were negative for Actinomyces-like organisms, pus cells were present in 8.7%, and yeast cells in 4.8%. Of the 7 smears positive for the organisms, only 1 was from the user of a plastic device. All cases included were asymptomatic and were visiting the clinic for routine follow-up. These data suggest that colonization by Actinomyces-like organisms does not appear to be a significant problem in IUD users in Malaysia.

ρ resonance from the I = 1 ππ potential in lattice QCD

NASA Astrophysics Data System (ADS)

Kawai, Daisuke

2018-03-01

We calculate the phase shift for the I = 1 ππ scattering in 2+1 flavor lattice QCD at mπ = 410 MeV, using all-to-all propagators with the LapH smearing. We first investigate the sink operator independence of the I = 2 ππ scattering phase shift to estimate the systematics in the LapH smearing scheme in the HAL QCD method at mπ = 870 MeV. The difference in the scattering phase shift in this channel between the conventional point sink scheme and the smeared sink scheme is reasonably small as long as the next-toleading analysis is employed in the smeared sink scheme with larger smearing levels. We then extract the I = 1 ππ potential with the smeared sink operator, whose scattering phase shift shows a resonant behavior (ρ resonance). We also examine the pole of the S-matrix corresponding to the ρ resonance in the complex energy plane.

A Prospective Study of Tuberculosis Drug Susceptibility in Sabah, Malaysia, and an Algorithm for Management of Isoniazid Resistance

PubMed Central

Rashid Ali, Muhammad Redzwan S.; Parameswaran, Uma; William, Timothy; Bird, Elspeth; Wilkes, Christopher S.; Lee, Wai Khew; Yeo, Tsin Wen; Anstey, Nicholas M.; Ralph, Anna P.

2015-01-01

Introduction. The burden of tuberculosis is high in eastern Malaysia, and rates of Mycobacterium tuberculosis drug resistance are poorly defined. Our objectives were to determine M. tuberculosis susceptibility and document management after receipt of susceptibility results. Methods. Prospective study of adult outpatients with smear-positive pulmonary tuberculosis (PTB) in Sabah, Malaysia. Additionally, hospital clinicians accessed the reference laboratory for clinical purposes during the study. Results. 176 outpatients were enrolled; 173 provided sputum samples. Mycobacterial culture yielded M. tuberculosis in 159 (91.9%) and nontuberculous Mycobacterium (NTM) in three (1.7%). Among outpatients there were no instances of multidrug resistant M. tuberculosis (MDR-TB). Seven people (4.5%) had isoniazid resistance (INH-R); all were switched to an appropriate second-line regimen for varying durations (4.5–9 months). Median delay to commencement of the second-line regimen was 13 weeks. Among 15 inpatients with suspected TB, 2 had multidrug resistant TB (one extensively drug resistant), 2 had INH-R, and 4 had NTM. Conclusions. Current community rates of MDR-TB in Sabah are low. However, INH-resistance poses challenges, and NTM is an important differential diagnosis in this setting, where smear microscopy is the usual diagnostic modality. To address INH-R management issues in our setting, we propose an algorithm for the treatment of isoniazid-resistant PTB. PMID:25838829

A prospective study of tuberculosis drug susceptibility in sabah, malaysia, and an algorithm for management of isoniazid resistance.

PubMed

Rashid Ali, Muhammad Redzwan S; Parameswaran, Uma; William, Timothy; Bird, Elspeth; Wilkes, Christopher S; Lee, Wai Khew; Yeo, Tsin Wen; Anstey, Nicholas M; Ralph, Anna P

2015-01-01

Introduction. The burden of tuberculosis is high in eastern Malaysia, and rates of Mycobacterium tuberculosis drug resistance are poorly defined. Our objectives were to determine M. tuberculosis susceptibility and document management after receipt of susceptibility results. Methods. Prospective study of adult outpatients with smear-positive pulmonary tuberculosis (PTB) in Sabah, Malaysia. Additionally, hospital clinicians accessed the reference laboratory for clinical purposes during the study. Results. 176 outpatients were enrolled; 173 provided sputum samples. Mycobacterial culture yielded M. tuberculosis in 159 (91.9%) and nontuberculous Mycobacterium (NTM) in three (1.7%). Among outpatients there were no instances of multidrug resistant M. tuberculosis (MDR-TB). Seven people (4.5%) had isoniazid resistance (INH-R); all were switched to an appropriate second-line regimen for varying durations (4.5-9 months). Median delay to commencement of the second-line regimen was 13 weeks. Among 15 inpatients with suspected TB, 2 had multidrug resistant TB (one extensively drug resistant), 2 had INH-R, and 4 had NTM. Conclusions. Current community rates of MDR-TB in Sabah are low. However, INH-resistance poses challenges, and NTM is an important differential diagnosis in this setting, where smear microscopy is the usual diagnostic modality. To address INH-R management issues in our setting, we propose an algorithm for the treatment of isoniazid-resistant PTB.

Morphological alterations of radicular dentine pretreated with different irrigating solutions and irradiated with 980-nm diode laser.

PubMed

Alfredo, Edson; Souza-Gabriel, Aline E; Silva, Silvio Rocha C; Sousa-Neto, Manoel D; Brugnera-Junior, Aldo; Silva-Sousa, Yara T C

2009-01-01

The topographical features of intraradicular dentine pretreated with sodium hypochlorite (NaOCl) or ethylenediamine tetraacetic acid (EDTA) followed by diode laser irradiation have not yet been determined. To evaluate the alterations of dentine irradiated with 980-nm diode laser at different parameters after the surface treatment with NaOCl and EDTA. Roots of 60 canines were biomechanically prepared and irrigated with NaOCl or EDTA. Groups were divided according to the laser parameters: 1.5 W/CW; 1.5 W/100 Hz; 3.0 W/CW; 3.0 W/100 Hz and no irradiation (control). The roots were splited longitudinally and analyzed by scanning electron microscopy (SEM) in a quali-quatitative way. The scores were submitted to two-way Kruskal-Wallis and Dunn's tests. The statistical analysis demonstrated that the specimens treated only with NaOCl or EDTA (control groups) were statistically different (P < 0.05) from the laser-irradiated specimens, regardless of the parameter setting. The specimens treated with NaOCl showed a laser-modified surface with smear layer, fissures, and no visible tubules. Those treated with EDTA and irradiated by laser presented absence of smear layer, tubules partially exposed and melting areas. The tested parameters of 980-nm diode laser promoted similar alterations on dentine morphology, dependent to the type of surface pretreatment. Copyright 2008 Wiley-Liss, Inc.

The efficacy of selective calculus ablation at 400 nm: comparison to conventional calculus removal methods

NASA Astrophysics Data System (ADS)

Schoenly, Joshua E.; Seka, Wolf; Romanos, Georgios; Rechmann, Peter

A desired outcome of scaling and root planing is the complete removal of calculus and infected root tissue and preservation of healthy cementum for rapid healing of periodontal tissues. Conventional periodontal treatments for calculus removal, such as hand instrument scaling and ultrasonic scaling, often deeply scrape the surface of the underlying hard tissue and may leave behind a smear layer. Pulsed lasers emitting at violet wavelengths (specifically, 380 to 400 nm) are a potential alternative treatment since they can selectively ablate dental calculus without ablating pristine hard tissue (i.e., enamel, cementum, and dentin). In this study, light and scanning electron microscopy are used to compare and contrast the efficacy of in vitro calculus removal for several conventional periodontal treatments (hand instruments, ultrasonic scaler, and Er:YAG laser) to calculus removal with a frequency-doubled Ti:sapphire (λ = 400 nm). After calculus removal, enamel and cementum surfaces are investigated for calculus debris and damage to the underlying hard tissue surface. Compared to the smear layer, grooves, and unintentional hard tissue removal typically found using these conventional treatments, calculus removal using the 400-nm laser is complete and selective without any removal of pristine dental hard tissue. Based on these results, selective ablation from the 400-nm laser appears to produce a root surface that would be more suitable for successful healing of periodontal tissues.

Root planing with Er:YAG laser X Gracey curette: a study in vitro using scanning electron microscopy

NASA Astrophysics Data System (ADS)

Mello, Fabiano A. S.; Mello, Andrea M. D.; Matson, Edmir; Mattos, Adriana B.; Mello, Guilherme P. S.

2001-04-01

The Er:YAG laser has been studied for periodontal therapies, so much for removal of the subgingival calculus and its bactericidal effects. The proposal of this study is to evaluate the effectiveness of the Er:YAG laser in root planning in comparison to the traditional method, not surgical. Six recently extracted due top the disease periodontal, were cut longitudinally tends like this two half of the root. These half were separate in four groups. The first group is the natural tooth, the second group was accomplished to rot planing with Graceycurette. And in the third to Er:YAG laser with a contact tip, using a 45 degree angle in relation to the root; in the fourth group was scraped and planed with Er:YAG laser and complemented rot planing with Graceycurette. The used energy was of 60 to 300mJ and the frequency of 10 Hz accomplished with irrigation. The obtained results were similar in the groups 2 and 3 in comparison to the amount of smear-layer. In group 4 however, better result was obtained, because the image was much more regular and with less amount of smear-layer. The conclusion of the work is that with the association of the Er:YAG laser technique and Graceycurette the results are superior to the conventional treatment.

Hematology, Parasitology, and Serology of Free-Ranging Coyotes (Canis latrans) from South Carolina.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miller, Debra, Lee; Schrecengost, Joshua; Merrill, Anita

2009-07-01

ABSTRACT: Blood and feces were collected from 34 adult (19 males, 15 females) and seven juvenile (three males, one female, three not reported) free-ranging coyotes (Canis latrans) on the US Department of Energy’s Savannah River Site (South Carolina, USA). Significant (P,0.05) hematologic differences by sex were noted for red blood cell counts, hemoglobin, and hematocrit. Biochemical differences by sex occurred only for albumen (P,0.05). Twentyone adults were antibody positive for at least one of four viruses: canine adenovirus type 1 (CAV-1; 68%), West Nile virus (WNV; 60%), Eastern equine encephalitis virus (EEEV; 38%), and Canine distemper virus (CDV; 15%). Ofmore » the seven Leptospira serovars tested for, seven (25%) of 28 adults were positive for one or more of five serovars: Pomona, Grippotyphosa, Icterohaemorrhagiae, Bratislava, and Autumnalis. Three (43%) of seven juveniles had seropositivity for a virus, one each for CDV, CAV-1, and WNV. No juveniles were seropositive for EEEV or any of the seven Leptospira serovars. Blood smears of 12 adults were positive for Dirofilaria immitis microfilaria, but blood smears from all juveniles were negative. Parvovirus was identified by electron microscopy from the feces of one adult. Ancylostoma spp., Trichuris spp., and Isospora spp. were observed in fecal samples. These data may aid in understanding the role of coyotes in disease ecology.« less

Trypanosoma cruzi infection in captive Neotropical primates in the Brazilian Amazon.

PubMed

Bahia, Michele; de Nazaré Leite Barros, Flávia; Magalhães-Matos, Paulo Cesar; de Souza Gonçalves, Thamirys; Chiesorin Neto, Laerzio; Oliveira Faria, Diogo Cesar Lagroteria; Aparecida Romeiro, Sandra; Barros Monteiro, Frederico Ozanan; Góes-Cavalcante, Gustavo; Scofield, Alessandra

2017-02-01

The aim of this study was to detect the infection by Trypanosoma cruzi in captive Neotropical primates in the Brazilian Amazon. From February 2013 to July 2014, 112 blood samples were collected from Neotropical primates from the Amazonas, Amapá, and Pará States, north of Brazil. The subjects belonged to the families Cebidae (N = 59), Atelidae (N = 41), Callitrichidae (N = 5), Pitheciidae (N = 4), and Aotidae (N = 3). Blood smears also were examined for the presence of trypomastigotes by optical microscopy. For the detection of T. cruzi DNA, a Nested-PCR with primers TCZ1/TCZ2 and TCZ3/TCZ4 was performed. T. cruzi DNA was detected in 12.5% (14/112) of Neotropical primates examined. Positive samples were detected in 16%, 12.5%, and 11.11% of the different species of primates sampled from the Amapá, Pará, and Amazonas states, respectively. The analysis of the blood smears did not reveal trypomastigote forms of T. cruzi. In conclusion, Neotropical primates kept in captivity were infected by T. cruzi in the studied areas. We recommend that a health management protocol be put into place to prevent the transmission of infectious agents among captive populations, captive and wild populations, and between NHPs and the technicians who handle these animals. © 2016 Wiley Periodicals, Inc.

The use of infrared thermography to detect the stages of estrus cycle and ovulation time in anatolian shepherd dogs.

PubMed

Olğaç, Kemal Tuna; Akçay, Ergun; Çil, Beste; Uçar, Burak Mehmet; Daşkın, Ali

2017-01-01

The aim of the study is to evaluate the effectiveness of thermographic monitoring, using the temperature changes of perianal and perivulvar areas for the determination of estrus in Anatolian Shepherd bitches. Fifteen bitches were used in the study. Blood and vaginal smear samples were collected and thermographic monitoring of perianal and perivulvar areas were carried out starting from proestrus to early diestrus. Also, external signs of estrus were investigated. Smear samples were evaluated by light microscopy after Diff-Quik staining method and superficial and keratinized superficial cells were determined as percentage (S + KS%). Progesterone and luteinizing hormone measurements were done by radioimmunoassay. The difference in temperature between perianal and perivulvar areas was evaluated through thermographic images by FLIR ResearchIR Software. According to the results obtained from the study, differences between progesterone and S + KS% were statistically significant ( P  < 0,05). Although temperature showed increase and decrease with progesterone and S + KS%, the differences were not important statistically ( P  > 0,05). Serum luteinizing hormone levels did not sign any difference ( P  > 0,05). As a result, thermographic monitoring alone is not enough for estrus detection in Anatolian Shepherd bitches. However, it can be used to assist the actual estrus detection technique in terms of providing some foreknowledge by evaluating the differences in temperature.

Performance of Rapid Diagnostic Tests for Imported Malaria in Clinical Practice: Results of a National Multicenter Study

PubMed Central

Houzé, Sandrine; Boutron, Isabelle; Marmorat, Anne; Dalichampt, Marie; Choquet, Christophe; Poilane, Isabelle; Godineau, Nadine; Le Guern, Anne-Sophie; Thellier, Marc; Broutier, Hélène; Fenneteau, Odile; Millet, Pascal; Dulucq, Stéphanie; Hubert, Véronique; Houzé, Pascal; Tubach, Florence; Le Bras, Jacques; Matheron, Sophie

2013-01-01

We compared the performance of four rapid diagnostic tests (RDTs) for imported malaria, and particularly Plasmodium falciparum infection, using thick and thin blood smears as the gold standard. All the tests are designed to detect at least one protein specific to P. falciparum ( Plasmodium histidine-rich protein 2 (PfHRP2) or Plasmodium LDH (PfLDH)) and one pan-Plasmodium protein (aldolase or Plasmodium LDH (pLDH)). 1,311 consecutive patients presenting to 9 French hospitals with suspected malaria were included in this prospective study between April 2006 and September 2008. Blood smears revealed malaria parasites in 374 cases (29%). For the diagnosis of P. falciparum infection, the three tests detecting PfHRP2 showed high and similar sensitivity (96%), positive predictive value (PPV) (90%) and negative predictive value (NPV) (98%). The PfLDH test showed lower sensitivity (83%) and NPV (80%), despite good PPV (98%). For the diagnosis of non-falciparum species, the PPV and NPV of tests targeting pLDH or aldolase were 94–99% and 52–64%, respectively. PfHRP2-based RDTs are thus an acceptable alternative to routine microscopy for diagnosing P. falciparum malaria. However, as malaria may be misdiagnosed with RDTs, all negative results must be confirmed by the reference diagnostic method when clinical, biological or other factors are highly suggestive of malaria. PMID:24098699

Gardnerella vaginalis and Lactobacillus sp in liquid-based cervical samples in healthy and disturbed vaginal flora using cultivation-independent methods.

PubMed

Klomp, Johanna M; Verbruggen, Banut-Sabine M; Korporaal, Hans; Boon, Mathilde E; de Jong, Pauline; Kramer, Gerco C; van Haaften, Maarten; Heintz, A Peter M

2008-05-01

Our objective was to determine the morphotype of the adherent bacteria in liquid-based cytology (LBC) in smears with healthy and disturbed vaginal flora. And to use PCR technology on the same fixed cell sample to establish DNA patterns of the 16S RNA genes of the bacteria in the sample. Thirty samples were randomly selected from a large group of cervical cell samples suspended in a commercial coagulant fixative "(BoonFix)." PCR was used to amplify DNA of five bacterial species: Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus jensenii, Gardnerella vaginalis, and Mycoplasma hominis. The LBC slides were then analyzed by light microscopy to estimate bacterial adhesion. DNA of lactobacilli was detected in all cell samples. Seventeen smears showed colonization with Gardnerella vaginalis (range 2.6 x 10(2)-3.0 x 10(5) bacteria/mul BoonFix sample). Two cases were identified as dysbacteriotic with high DNA values for Gardnerella vaginalis and low values for Lactobacillus crispatus. The sample with the highest concentration for Gardnerella vaginalis showed an unequivocal Gardnerella infection. This study indicates that the adherence pattern of a disturbed flora in liquid-based cervical samples can be identified unequivocally, and that these samples are suitable for quantitative PCR analysis. This cultivation independent method reveals a strong inverse relationship between Gardnerella vaginalis and Lactobacillus crispatus in dysbacteriosis and unequivocal Gardnerella infection.

An audit of a cervical smear screening programme.

PubMed

Moodie, P J; Kljakovic, M; McLeod, D K

1989-07-26

An audit of a computer based screening and recall programme in a Wellington group general practice is reported (practice population 13,866). The records of all women aged between 20 and 59 years (4133 women) were checked to determine if they had had a cervical smear test in the previous two years. A random sample of women who had a cervical smear result recorded in the notes (107 women called "responders") showed that 71% gave "familiarity with the family doctor" and "acting in response to a recall letter" as reasons for choosing the place of their last smear. Satisfaction with the service was indicated by 95% of these women stating they would have their next smear at the medical centre. In the audit of all the records, a group of 667 women who had been sent a letter inviting them to have a smear done and who had apparently declined the procedure was identified (called "nonresponders"). A random sample of this group (168 women) was taken and an attempt made to interview them. In fact only 38 women could be identified as requiring a smear and even if those who refused to be interviewed (13) and those unable to be contacted (23) are added, then less than half of this sample were "true nonresponders". This suggests that the percentage of women in the practice who have been offered a smear and have refused to have one is less than 8%.

[Accuracy of three methods for the rapid diagnosis of oral candidiasis].

PubMed

Lyu, X; Zhao, C; Yan, Z M; Hua, H

2016-10-09

Objective: To explore a simple, rapid and efficient method for the diagnosis of oral candidiasis in clinical practice. Methods: Totally 124 consecutive patients with suspected oral candidiasis were enrolled from Department of Oral Medicine, Peking University School and Hospital of Stomatology, Beijing, China. Exfoliated cells of oral mucosa and saliva or concentrated oral rinse) obtained from all participants were tested by three rapid smear methods(10% KOH smear, gram-stained smear, Congo red stained smear). The diagnostic efficacy(sensitivity, specificity, Youden's index, likelihood ratio, consistency, predictive value and area under curve(AUC) of each of the above mentioned three methods was assessed by comparing the results with the gold standard(combination of clinical diagnosis, laboratory diagnosis and expert opinion). Results: Gram-stained smear of saliva(or concentrated oral rinse) demonstrated highest sensitivity(82.3%). Test of 10%KOH smear of exfoliated cells showed highest specificity(93.5%). Congo red stained smear of saliva(or concentrated oral rinse) displayed highest diagnostic efficacy(79.0% sensitivity, 80.6% specificity, 0.60 Youden's index, 4.08 positive likelihood ratio, 0.26 negative likelihood ratio, 80% consistency, 80.3% positive predictive value, 79.4% negative predictive value and 0.80 AUC). Conclusions: Test of Congo red stained smear of saliva(or concentrated oral rinse) could be used as a point-of-care tool for the rapid diagnosis of oral candidiasis in clinical practice. Trial registration: Chinese Clinical Trial Registry, ChiCTR-DDD-16008118.

Identification of mycobacterium tuberculosis in sputum smear slide using automatic scanning microscope

NASA Astrophysics Data System (ADS)

Rulaningtyas, Riries; Suksmono, Andriyan B.; Mengko, Tati L. R.; Saptawati, Putri

2015-04-01

Sputum smear observation has an important role in tuberculosis (TB) disease diagnosis, because it needs accurate identification to avoid high errors diagnosis. In development countries, sputum smear slide observation is commonly done with conventional light microscope from Ziehl-Neelsen stained tissue and it doesn't need high cost to maintain the microscope. The clinicians do manual screening process for sputum smear slide which is time consuming and needs highly training to detect the presence of TB bacilli (mycobacterium tuberculosis) accurately, especially for negative slide and slide with less number of TB bacilli. For helping the clinicians, we propose automatic scanning microscope with automatic identification of TB bacilli. The designed system modified the field movement of light microscope with stepper motor which was controlled by microcontroller. Every sputum smear field was captured by camera. After that some image processing techniques were done for the sputum smear images. The color threshold was used for background subtraction with hue canal in HSV color space. Sobel edge detection algorithm was used for TB bacilli image segmentation. We used feature extraction based on shape for bacilli analyzing and then neural network classified TB bacilli or not. The results indicated identification of TB bacilli that we have done worked well and detected TB bacilli accurately in sputum smear slide with normal staining, but not worked well in over staining and less staining tissue slide. However, overall the designed system can help the clinicians in sputum smear observation becomes more easily.

Local spatial variations analysis of smear-positive tuberculosis in Xinjiang using Geographically Weighted Regression model.

PubMed

Wei, Wang; Yuan-Yuan, Jin; Ci, Yan; Ahan, Alayi; Ming-Qin, Cao

2016-10-06

The spatial interplay between socioeconomic factors and tuberculosis (TB) cases contributes to the understanding of regional tuberculosis burdens. Historically, local Poisson Geographically Weighted Regression (GWR) has allowed for the identification of the geographic disparities of TB cases and their relevant socioeconomic determinants, thereby forecasting local regression coefficients for the relations between the incidence of TB and its socioeconomic determinants. Therefore, the aims of this study were to: (1) identify the socioeconomic determinants of geographic disparities of smear positive TB in Xinjiang, China (2) confirm if the incidence of smear positive TB and its associated socioeconomic determinants demonstrate spatial variability (3) compare the performance of two main models: one is Ordinary Least Square Regression (OLS), and the other local GWR model. Reported smear-positive TB cases in Xinjiang were extracted from the TB surveillance system database during 2004-2010. The average number of smear-positive TB cases notified in Xinjiang was collected from 98 districts/counties. The population density (POPden), proportion of minorities (PROmin), number of infectious disease network reporting agencies (NUMagen), proportion of agricultural population (PROagr), and per capita annual gross domestic product (per capita GDP) were gathered from the Xinjiang Statistical Yearbook covering a period from 2004 to 2010. The OLS model and GWR model were then utilized to investigate socioeconomic determinants of smear-positive TB cases. Geoda 1.6.7, and GWR 4.0 software were used for data analysis. Our findings indicate that the relations between the average number of smear-positive TB cases notified in Xinjiang and their socioeconomic determinants (POPden, PROmin, NUMagen, PROagr, and per capita GDP) were significantly spatially non-stationary. This means that in some areas more smear-positive TB cases could be related to higher socioeconomic determinant regression coefficients, but in some areas more smear-positive TB cases were found to do with lower socioeconomic determinant regression coefficients. We also found out that the GWR model could be better exploited to geographically differentiate the relationships between the average number of smear-positive TB cases and their socioeconomic determinants, which could interpret the dataset better (adjusted R 2  = 0.912, AICc = 1107.22) than the OLS model (adjusted R 2  = 0.768, AICc = 1196.74). POPden, PROmin, NUMagen, PROagr, and per capita GDP are socioeconomic determinants of smear-positive TB cases. Comprehending the spatial heterogeneity of POPden, PROmin, NUMagen, PROagr, per capita GDP, and smear-positive TB cases could provide valuable information for TB precaution and control strategies.

[Quality assessment of microscopic examination in tuberculosis diagnostic laboratories: a preliminary study].

PubMed

Simşek, Hülya; Ceyhan, Ismail; Tarhan, Gülnur; Güner, Uğur

2010-10-01

Recently, the diagnosis of pulmonary tuberculosis (TB) has based on smear microscopy in the Direct Observed Treatment Strategy (DOTS) programme which provides the basis of treatment worldwide. Microscopic detection of AFB (Acid-Fast Bacilli) is one of the main components in the National TB Control Programmes (NTCP). Precision level in microscopy procedures and evaluations are the most important steps for accurate diagnosis of the disease and to initiate proper treatment. Therefore, the external quality assessment (EQA) is the most important implement to provide the reliability and validity of tests. In countries where NTCP are performed, this task is fulfilled by the National Reference Laboratories (NRL) according to the guidelines of the World Health Organization (WHO). For this purpose a pilot study was initiated by the central NRL of Turkey for EQA of AFB smear microscopy as part of the NTCP on January 1, 2005. A total of 5 laboratories of which 2 were district TB laboratories (A, B), 2 were tuberculosis control dispensaries (C, D), 1 was a national reference laboratory (E), participated in this study. Blind re-checking method (re-examination of randomly selected slides) was used for the evaluation, and the slides were sent to the central NRL with 3 months interval, four times a year, selected according to LQAS (Lot Quality Assurance Sampling) guides. In the re-evaluation of the slides, false positivity (FP), false negativity (FN) and quantification errors (QE) were noted. Laboratory A, sent totally 525 slides between January 1, 2005 and April 1, 2008. In the result of re-checking, 514 (97.9%) slides were found concordant, and 11 (2.1%) were discordant (10 FP, 1 FN). Laboratory B, participated in the study between October 1, 2005 and July 1, 2006 and of the 67 re-examined slides, 60 (89.5%) were concordant and 7 (10.5%) were discordant (2 FP, 0 FN, 5 QE). Laboratory C, sent 235 slides between January 1, 2005 and April 1, 2006; of them 218 (92.8%) were detected as compatible and 17 (7.2%) slides were incompatible (4 FP, 9 FN, 4 QE). Laboratory D, participated in QC for only once between January 1, 2008 and April 1, 2008; and all the 50 slides were found compatible, with no FP, FN and QE. Laboratory E, was included in the study between January 1, 2005 and January 1, 2008 and of the 696 re-checked slides, 690 (99.1%) were reported as compatible and 6 (0.9%) were incompatible (3 FN, 3 QE). Following EQA, on-site evaluation of the laboratories with major errors, was performed and necessary adjustments and training were done. In conclusion, external quality control measures for AFB microscopy is crucial and essential for the tuberculosis laboratory performances for accurate and reliable results.

Analysis of small-angle X-ray scattering data in the presence of significant instrumental smearing

PubMed Central

Bergenholtz, Johan; Ulama, Jeanette; Zackrisson Oskolkova, Malin

2016-01-01

A laboratory-scale small-angle X-ray scattering instrument with pinhole collimation has been used to assess smearing effects due to instrumental resolution. A new, numerically efficient method to smear ideal model intensities is developed and presented. It allows for directly using measured profiles of isotropic but otherwise arbitrary beams in smearing calculations. Samples of low-polydispersity polymer spheres have been used to show that scattering data can in this way be quantitatively modeled even when there is substantial distortion due to instrumental resolution. PMID:26937235

Analytical solution for vacuum preloading considering the nonlinear distribution of horizontal permeability within the smear zone.

PubMed

Peng, Jie; He, Xiang; Ye, Hanming

2015-01-01

The vacuum preloading is an effective method which is widely used in ground treatment. In consolidation analysis, the soil around prefabricated vertical drain (PVD) is traditionally divided into smear zone and undisturbed zone, both with constant permeability. In reality, the permeability of soil changes continuously within the smear zone. In this study, the horizontal permeability coefficient of soil within the smear zone is described by an exponential function of radial distance. A solution for vacuum preloading consolidation considers the nonlinear distribution of horizontal permeability within the smear zone is presented and compared with previous analytical results as well as a numerical solution, the results show that the presented solution correlates well with the numerical solution, and is more precise than previous analytical solution.

Analytical solution for vacuum preloading considering the nonlinear distribution of horizontal permeability within the smear zone

PubMed Central

Peng, Jie; He, Xiang; Ye, Hanming

2015-01-01

The vacuum preloading is an effective method which is widely used in ground treatment. In consolidation analysis, the soil around prefabricated vertical drain (PVD) is traditionally divided into smear zone and undisturbed zone, both with constant permeability. In reality, the permeability of soil changes continuously within the smear zone. In this study, the horizontal permeability coefficient of soil within the smear zone is described by an exponential function of radial distance. A solution for vacuum preloading consolidation considers the nonlinear distribution of horizontal permeability within the smear zone is presented and compared with previous analytical results as well as a numerical solution, the results show that the presented solution correlates well with the numerical solution, and is more precise than previous analytical solution. PMID:26447973

The Sensitivity and Specificity of Potassium Hydroxide Smear and Fungal Culture Relative to Clinical Assessment in the Evaluation of Tinea Pedis: A Pooled Analysis

PubMed Central

Levitt, Jacob Oren; Levitt, Barrie H.; Akhavan, Arash; Yanofsky, Howard

2010-01-01

Background. There are relatively few studies published examining the sensitivity and specificity of potassium hydroxide (KOH) smear and fungal culture examination of tinea pedis. Objective. To evaluate the sensitivity and specificity of KOH smear and fungal culture for diagnosing tinea pedis. Methods. A pooled analysis of data from five similarly conducted bioequivalence trials for antifungal drugs was performed. Data from 460 patients enrolled in the vehicle arms of these studies with clinical diagnosis of tinea pedis supported by positive fungal culture were analyzed 6 weeks after initiation of the study to determine the sensitivity and specificity of KOH smear and fungal culture. Results. Using clinical assessment as the gold standard, the sensitivities for KOH smear and culture were 73.3% (95% CI: 66.3 to 79.5%) and 41.7% (34.6 to 49.1%), respectively. The respective specificities for culture and KOH smear were 77.7% (72.2 to 82.5%) and 42.5% (36.6 to 48.6%). Conclusion. KOH smear and fungal culture are complementary diagnostic tests for tinea pedis, with the former being the more sensitive test of the two, and the latter being more specific. PMID:20672004

Inflammation on the Cervical Papanicolaou Smear: Evidence for Infection in Asymptomatic Women?

PubMed Central

Tsirmpa, Ioanna; Chasiakou, Anthia; Tsouma, Iliana; Politi, Ekaterina; Gennimata, Vassiliki; Kouskouni, Evangelia

2013-01-01

Background. The significance of the possible presence of infection on the Pap smear of asymptomatic women based on cytological criteria is practically unknown. Materials and Methods. A total of 1117 asymptomatic nonpregnant women had Pap smear tests and vaginal as well as cervical cultures completed (622 with and 495 without inflammation on the Pap smear). Results. Out of the 622 women with inflammation on Pap test, 251 (40.4%) had negative cultures (normal flora present), while 371 (59.6%) women had positive cultures with different pathogens. In contrast, the group of women without inflammation on Pap test displayed significantly increased percentage of negative cultures (67.1%, P < 0.001) and decreased percentage of positive cultures (32.9%, P < 0.001). Bacterial vaginosis was diagnosed more frequently in both groups and significantly more in the group with inflammation on Pap smear compared to the group without inflammation (P < 0.02). Conclusions. A report of inflammatory changes on the cervical Pap smear cannot be used to reliably predict the presence of a genital tract infection, especially in asymptomatic women. Nevertheless, the isolation of different pathogens in about 60% of the women with inflammation on the Pap smear cannot be overlooked and must be regarded with concern. PMID:24204103

Cervical cancer screening after 50: near extinction?

PubMed

Abdullah, Nik N; Daud, Suzanna; Al-Kubaisy, Waqar; Saari, Izni S; Saad, Siti R

2016-11-01

The objectives of this study were to determine the prevalence and predictors of Pap smear screening among women aged 50 years and older. This cross-sectional study was conducted at two large urban health centres in Selangor. A total of 515 women aged 50 and older were recruited. The mean age of the respondents was 58.83±7.05, with a range of 50- 83 years. The prevalence of Pap smear screening was 39.22% (n=202). From the multivariate analysis, Pap smear screening was significantly associated with health care provider advice (adjusted odds ratio (AOR)=18.75; 95% CI=8.30, 42.37); tertiary (AOR=11.26; 95% CI=1.50, 84.68) and secondary education level (AOR=9.47; 95% CI=1.43, 62.84); use of contraception (AOR=2.90; 95% CI=1.48, 5.69); heart disease (AOR=0.22; 95% CI=0.05, 0.97); and worry about Pap smear results (AOR=0.20; 95% CI=0.09, 0.42). The prevalence of Pap smear screening in the older women is unsatisfactory. Health care provider advice, education level, use of contraception, heart disease and worry about Pap smear results were predictors of undergoing Pap smear screening in this study population. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Thick Smear is a Good Substitute for the Thin Smear in Parasitological Confirmation of Canine Visceral Leishmaniasis.

PubMed

de Mello, Cintia Xavier; Figueiredo, Fabiano Borges; Mendes Júnior, Artur Augusto Velho; Miranda, Luciana de Freitas Campos; de Oliveira, Raquel de Vasconcellos Carvalhaes; Madeira, Maria de Fátima

2016-07-06

Although direct examination methods are important for diagnosing leishmaniasis, such methods are often neglected because of their low sensitivity relative to other techniques. Our study aimed to evaluate the performance of bone marrow (BM) thick smears and cytocentrifugation tests as alternatives to direct examination for diagnosing canine visceral leishmaniasis (CVL). Ninety-two dogs exhibiting leishmaniasis seroreactivity were evaluated. The animals were euthanized; and healthy skin, spleen, popliteal lymph node, and BM puncture samples were cultured. BM cultures were used as the reference standard. Of the 92 dogs studied, 85.9% exhibited positive cultures, and Leishmania infantum (synonym Leishmania chagasi) was confirmed in all positive culture cases. The sensitivity rates for cytocentrifugation as well as thin and thick smears were 47.1%, 52.8%, and 77%, respectively. However, no association between the dogs' clinical status and culture or direct examination results was found. To our knowledge, this was the first study to use thick smears and cytocentrifugation for diagnosing CVL. Our results indicate that BM thick smears have a good sensitivity and their use reduces the time required to read slides. Therefore, thick smears can provide a rapid and safe alternative to parasitological confirmation of seroreactive dogs. © The American Society of Tropical Medicine and Hygiene.

Evaluation of the ICT Tuberculosis test for the routine diagnosis of tuberculosis

PubMed Central

Ongut, Gozde; Ogunc, Dilara; Gunseren, Filiz; Ogus, Candan; Donmez, Levent; Colak, Dilek; Gultekin, Meral

2006-01-01

Background Rapid and accurate diagnosis of tuberculosis (TB) is crucial to facilitate early treatment of infectious cases and thus to reduce its spread. To improve the diagnosis of TB, more rapid diagnostic techniques such as antibody detection methods including enzyme-linked immunosorbent assay (ELISA)-based serological tests and immunochromatographic methods were developed. This study was designed to evaluate the validity of an immunochromatographic assay, ICT Tuberculosis test for the serologic diagnosis of TB in Antalya, Turkey. Methods Sera from 72 patients with active pulmonary (53 smear-positive and 19 smear-negative cases) and eight extrapulmonary (6 smear-positive and 2 smear-negative cases) TB, and 54 controls from different outpatient clinics with similar demographic characteristics as patients were tested by ICT Tuberculosis test. Results The sensitivity, specificity, and negative predictive value of the ICT Tuberculosis test for pulmonary TB were 33.3%, 100%, and 52.9%, respectively. Smear-positive pulmonary TB patients showed a higher positivity rate for antibodies than smear-negative patients, but the difference was not statistically significant. Of the eight patients with extrapulmonary TB, antibody was detected in four patients. Conclusion Our results suggest that ICT Tuberculosis test can be used to aid TB diagnosis in smear-positive patients until the culture results are available. PMID:16504161

Inflammatory events as detected in cervical smears and squamous intraepithelial lesions.

PubMed

Roeters, Anne M E; Boon, Mathilde E; van Haaften, Maarten; Vernooij, F; Bontekoe, Tj Romke; Heintz, A Peter M

2010-02-01

The Dutch cytological coding system, KOPAC, enables to code for eight inflammatory events, that is koilocytosis (related to human papillomavirus (HPV)), Trichomonas, dysbacteriosis [related to bacterial vaginosis (BV)], Candida, Gardnerella, Actinomyces, Chlamydia, and non-specific inflammation (leucocytosis). This study presents an analysis of 1,008,879 smears. Of each smear, the age of the woman and the reason for smear taking (screening or indication) was available. The cytoscores (per mille) for these codes were calculated. For the screening smears, the cytoscores were for koilocytosis (HPV) 2.6, for Trichomonas vaginalis 1.9, for dysbacteriosis 31.4, for Candida albicans 9.8, for Gardnerella vaginalis 0.7, for Actinomyces 6.9, for Chlamydia 0.8, and for non-specific inflammatory changes 66.4. For the calculation of the Odds Ratio (OR), normal smears were used as a reference. The cytoscores for Chlamydia and Gardnerella covaried with high grade SIL (HSIL), with an OR of 7 and 12, respectively. In addition, the OR for Trichomonas vaginalis, for dysbacteriosis, and for leucocytosis proved to be significantly high in the indication smears. This study provides an oversight of HSIL and the full range of cervical infections as detected by cytology, proving that this infectious byproduct of screening can be very valuable.

Efficacy of BACTEC TB in the rapid confirmatory diagnosis of mycobacterial infections. A Lebanese tertiary care center experience.

PubMed

Itani, Lina Y; Cherry, Mohamad A; Araj, George F

2005-01-01

Rapid detection of Mycobacterium tuberculosis (MTB), especially multidrug-resistant strains, is of importance for prompt clinical management and initiation of public health control measures. Culture remains the "gold" standard in the confirmatory laboratory diagnosis of mycobacterial infections. The reliability of the automated radiometric BACTEC 460 TB (BACTEC) system for the rapid detection of mycobacteria in clinical specimens was evaluated and compared to the conventional culture on Lowenstein-Jensen (LJ) medium. All clinical specimens submitted for mycobacterial culture were processed and simultaneously cultured on both BACTEC broth medium and LJ solid medium. Acid-fast bacilli (AFB) smears were also performed on the sediments. Differentiation of mycobacterial isolates as MTB or Mycobacterium sp. other than tuberculosis (MOTT) was based on the BACTEC NAP test. All positive culture findings recovered between January 1997 and December 2003 were analyzed in this study. A total of 3300 specimens were tested of which 355 (10.7%) yielded positive cultures consisting of 233 (65.6%) MTB and 122 (34.4%) MOTT. The percentages of AFB smear-positive were 45% and 49% in clinical specimens yielding MTB & MOTT, respectively. Though several types of specimens were cultured, most isolates (72% of MTB & 91% of MOTT) were recovered from respiratory specimens. Overall, the BACTEC showed significantly higher mycobacteria recovery rate (91%) than LJ (77%). In terms of times to detection, BACTEC showed significantly shorter detection time of isolates than LJ for the overall (mean 9.6 days for BACTEC vs. 22.8 days for LJ) and for each category of AFB smear finding. The detection time is shortened for BACTEC with the increasing grade of smear positivity. BACTEC is substantially more sensitive, efficient and rapid than LJ in the laboratory diagnosis of mycobacterial infections. This system also provides rapid differentiation of MTB from MOTT and susceptibility test results on MTB. However, the simultaneous use of BACTEC and LJ is recommended to provide maximum optimal recovery of isolates from clinical specimens. The time-saving in BACTEC provides an excellent facility for physicians in patient management and to public health personnel for prompt initiation of infection control measures.

Analysis of 13 million individual patient records pertaining to Pap smears, colposcopies, biopsies and surgery on the uterine cervix (Belgium, 1996-2000).

PubMed

Arbyn, Marc; Simoens, Cindy; Van Oyen, Herman; Foidart, Jean-Michel; Goffin, Frédéric; Simon, Philippe; Fabri, Valérie

2009-05-01

Cervical cancer screening by surveys overestimate coverage because of selection and reporting biases. The prepared Inter-Mutualistic Agency dataset has about 13 million records from Pap smears, colposcopies, cervical biopsies and surgery, performed in Belgium between 1996 and 2000. Cervical cancer screening coverage was defined as the proportion of the target population (women of 25-64 years) that has had a Pap smear taken within the last 3 years. Proportions and incidence rates were computed using official population data of the corresponding age group, area and calendar year. Cervical cancer screening coverage, in the period 1998-2000, was 59% at national level, for the target age group 25-64 years. Differences were small between the 3 regions. Variation ranged from 39% to 71%. Coverage was 64% for 25-29 year old women, 67% for those aged 30-39 years, 56% for those aged 50-54. The modal screening interval was 1 year. In the 3-year period 1998-2000, 3 million smears were taken from the 2.7 million women in the age group 25-64. Only 1.6 million women of the target group got one or more smears in that period and 1.1 million women had no smears, corresponding to an average of 1.88 smears per woman. Coverage reached only 59%, but the number of smears used was sufficient to cover more than 100% of the target population. Structural reduction of overuse and extension of coverage is warranted.

Diagnostic performance of automated liquid culture and molecular line probe assay in smear-negative pulmonary tuberculosis.

PubMed

Kotwal, Aarti; Biswas, Debasis; Raghuvanshi, Shailendra; Sindhwani, Girish; Kakati, Barnali; Sharma, Shweta

2017-04-01

The diagnosis of smear-negative pulmonary tuberculosis (PTB) is particularly challenging, and automated liquid culture and molecular line probe assays (LPA) may prove particularly useful. The objective of our study was to evaluate the diagnostic potential of automated liquid culture (ALC) technology and commercial LPA in sputum smear-negative PTB suspects. Spot sputum samples were collected from 145 chest-symptomatic smear-negative patients and subjected to ALC, direct drug susceptibility test (DST) testing and LPA, as per manufacturers' instructions. A diagnostic yield of 26.2% was observed among sputum smear-negative TB suspects with 47.4% of the culture isolates being either INH- and/or rifampicin-resistant. Complete agreement was observed between the results of ALC assay and LPA except for two isolates which demonstrated sensitivity to INH and rifampicin at direct DST but were rifampicin-resistant in LPA. Two novel mutations were also detected among the multidrug isolates by LPA. In view of the diagnostic challenges associated with the diagnosis of TB in sputum smear-negative patients, our study demonstrates the applicability of ALC and LPA in establishing diagnostic evidence of TB.

The effect of root surface conditioning on smear layer removal in periodontal regeneration (a scanning electron microscopic study)

NASA Astrophysics Data System (ADS)

Fidyawati, D.; Soeroso, Y.; Masulili, S. L. C.

2017-08-01

The role of root surface conditioning treatment on smear layer removal of human teeth is affected by periodontitis in periodontal regeneration. The objective of this study is to analyze the smear layer on root surface conditioned with 2.1% minocycline HCl ointment (Periocline), and 24% EDTA gel (Prefgel). A total of 10 human teeth indicated for extraction due to chronic periodontitis were collected and root planed. The teeth were sectioned in thirds of the cervical area, providing 30 samples that were divided into three groups - minocycline ointment treatment, 24% EDTA gel treatment, and saline as a control. The samples were examined by scanning electron microscope. No significant differences in levels of smear layer were observed between the minocycline group and the EDTA group (p=0.759). However, there were significant differences in the level of smear layer after root surface treatment in the minocycline and EDTA groups, compared with the control group (p=0.00). There was a relationship between root surface conditioning treatment and smear layer levels following root planing.

Immunofluorescence of bovine virus diarrhea viral antigen in white blood cells from experimentally infected immunocompetent calves.

PubMed Central

Bezek, D M; Baker, J C; Kaneene, J B

1988-01-01

A study to evaluate the detection of bovine virus diarrhea viral antigen using immunofluorescence testing of white blood cells was conducted. Five colostrum-deprived calves were inoculated intravenously with a cytopathic strain of the virus. Lymphocyte and buffy coat smears were prepared daily for direct immunofluorescent staining for detection of antigen. Lymphocytes were separated from heparinized blood using a Ficoll density procedure. Buffy coat smears were prepared from centrifuged blood samples collected using ethylenediaminetetraacetic acid as an anticoagulant. Bovine viral diarrhea virus antigen was detected by immunofluorescence between 3 and 11 days postinfection in lymphocyte smears and 3 to 12 days postinfection in buffy coat smears. Isolation of virus from both lymphocytes and buffy coat preparations correlated with detection of immunofluorescence. Serum neutralizing antibody to bovine virus diarrhea virus was detected on day 10 postinfection. Buffy coat smears were as sensitive as lymphocyte smears for the detection of antigen by immunofluorescence. It appeared that immunofluorescent staining of white blood cells was an effective method of detecting bovine virus diarrhea viral antigen. PMID:2836047

Resonance Raman microscopy in combination with partial dark-field microscopy lights up a new path in malaria diagnostics.

PubMed

Wood, Bayden R; Hermelink, Antje; Lasch, Peter; Bambery, Keith R; Webster, Grant T; Khiavi, Mehdi Asghari; Cooke, Brian M; Deed, Samantha; Naumann, Dieter; McNaughton, Don

2009-06-01

Our goal is to produce a rapid and accurate diagnostic tool for malaria using resonance Raman spectroscopy to detect small inclusions of haemozoin in Plasmodium falciparum infected red blood cells. In pursuit of this aim we serendipitously discovered a partial dark-field effect generated by our experimental setup, which helps identify in thick blood films potential parasites that are normally difficult to see with conventional bright-field microscopy. The haemozoin deposits 'light up' and these can be selectively targeted with the Raman microscope to confirm the presence or absence of haemozoin by the strong 1569 cm(-1) band, which is a marker for haemozoin. With newly developed imaging Raman microscopes incorporating ultra-sensitive rapid readout CCDs it is possible to obtain spectra with a good signal-to-noise ratio in 1 second. Moreover, images from a smear of potentially infected cells can be recorded and analysed with multivariate methods. The reconstructed images show what appear to be sub-micron-inclusions of haemozoin in some cells indicating that the technique has potential to identify low pigmented forms of the parasite including early trophozoite-stage infected cells. Further work is required to unambiguously confirm the presence of such forms through systematic staining but the results are indeed promising and may lead to the development of a new Raman-based malaria diagnostic.

DotLens smartphone microscopy for biological and biomedical applications (Conference Presentation)

NASA Astrophysics Data System (ADS)

Sung, Yu-Lung; Zhao, Fusheng; Shih, Wei-Chuan

2017-02-01

Recent advances in inkjet-printed optics have created a new class of lens fabrication technique. Lenses with a tunable geometry, magnification, and focal length can be fabricated by dispensing controlled amounts of liquid polymer onto a heated surface. This fabrication technique is highly cost-effective, and can achieve optically smooth surface finish. Dubbed DotLens, a single of which weighs less than 50 mg and occupies a volume less than 50 μL. DotLens can be attached onto any smartphone camera akin to a contact lens, and enable smartphones to obtain image resolution as fine as 1 µm. The surface curvature modifies the optical path of light to the image sensor, and enables the camera to focus as close as 2 mm. This enables microscopic imaging on a smartphone without any additional attachments, and has shown great potential in mobile point-of-care diagnostic systems, particularly for histology of tissue sections and cytology of blood cells. DotLens Smartphone Microscopy represents an innovative approach fundamentally different from other smartphone microscopes. In this paper, we describe the application and performance of DotLens smartphone microscopy in biological and biomedical research. In particular, we show recent results from images collected from pathology tissue slides with cancer features. In addition, we show performance in cytological analysis of blood smear. This tool has empowered Citizen Science investigators to collect microscopic images from various interesting objects.

Standard Operating Procedure Utilization for Tuberculosis Microscopy in Mekelle City, North Ethiopia.

PubMed

Weldu, Yemane; Gebru, Hagos; Kahsay, Getahun; Teweldemedhn, Gebremichael; Hagos, Yifter; Kahsay, Amlsha

2017-01-01

The aim of this study was to assess the utilization of standard operating procedures for acid-fast bacilli (AFB) smear microscopy. A facility-based cross-sectional study was conducted in select health institutions in Mekelle City, Ethiopia, from July 1, 2015, through August 30, 2015. Using a simple random sampling technique, 18 health facilities were included in the study. Data were collected using a standard checklist and entered into Epi Info version 3.5.4 (Centers for Disease Control and Prevention, Atlanta, GA) for editing. Analysis was done using SPSS version 20 (SPSS, Chicago, IL). Of the 18 laboratory facilities, only seven (38.9%) had a legible AFB registration book. In three (16.7%) of the laboratories, heat fixation was not applied before adding primary staining reagent. In 12 (66.7%), the staining reagents had precipitates. Two laboratories had microscopes with mechanical stages that could not move freely on both axes. Seven (38.9%) of the laboratories reported samples to be negative before examining all required fields. Most laboratories, 16 (88.9%) and 17 (94.4%), respectively, did not run positive and negative controls after new batch reagent preparation. Tuberculosis microscopy was found to be substandard with clear gaps in documentation, sample collection, and processing. © American Society for Clinical Pathology, 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Multilevel Optimization Framework for Hierarchical Stiffened Shells Accelerated by Adaptive Equivalent Strategy

NASA Astrophysics Data System (ADS)

Wang, Bo; Tian, Kuo; Zhao, Haixin; Hao, Peng; Zhu, Tianyu; Zhang, Ke; Ma, Yunlong

2017-06-01

In order to improve the post-buckling optimization efficiency of hierarchical stiffened shells, a multilevel optimization framework accelerated by adaptive equivalent strategy is presented in this paper. Firstly, the Numerical-based Smeared Stiffener Method (NSSM) for hierarchical stiffened shells is derived by means of the numerical implementation of asymptotic homogenization (NIAH) method. Based on the NSSM, a reasonable adaptive equivalent strategy for hierarchical stiffened shells is developed from the concept of hierarchy reduction. Its core idea is to self-adaptively decide which hierarchy of the structure should be equivalent according to the critical buckling mode rapidly predicted by NSSM. Compared with the detailed model, the high prediction accuracy and efficiency of the proposed model is highlighted. On the basis of this adaptive equivalent model, a multilevel optimization framework is then established by decomposing the complex entire optimization process into major-stiffener-level and minor-stiffener-level sub-optimizations, during which Fixed Point Iteration (FPI) is employed to accelerate convergence. Finally, the illustrative examples of the multilevel framework is carried out to demonstrate its efficiency and effectiveness to search for the global optimum result by contrast with the single-level optimization method. Remarkably, the high efficiency and flexibility of the adaptive equivalent strategy is indicated by compared with the single equivalent strategy.

The value of Tzanck smear test in diagnosis of erosive, vesicular, bullous, and pustular skin lesions.

PubMed

Durdu, Murat; Baba, Mete; Seçkin, Deniz

2008-12-01

Tzanck smear is generally used for the diagnosis of the pemphigus group of autoimmune bullous diseases and mucocutaneous herpesvirus infections. There are only a few studies in the literature investigating its diagnostic value. We aimed to investigate Tzanck smear findings and to determine the diagnostic value of this test in moist (erosive, vesicular, bullous, and pustular) skin lesions. We also aimed to develop an algorithmic approach for the diagnosis of these types of skin lesions according to the Tzanck smear findings. Samples were stained with May-Grünwald-Giemsa and evaluated by the same dermatologist. In some patients, methylene blue and Gram staining or direct immunofluorescence examinations were additionally performed. In all of the study cases, after the evaluation of clinical and laboratory findings (including, when appropriate, potassium hydroxide examination; viral serology; bacterial and fungal cultures; histopathology; direct and indirect immunofluorescence; patch testing), the definite diagnosis was established. We also determined the sensitivity and the specificity of certain Tzanck smear findings. Tzanck smear was performed in a total of 400 patients with moist skin lesions. The sensitivities of multinucleated giant cells and acantholytic cells in herpetic infections, dyskeratotic acantholytic cells and cocci in bullous impetigo, pseudohyphae in candidiasis, acantholytic cells in pemphigus and more than 10 tadpole cells (magnification x100) in spongiotic dermatitis were 84.7%, 92%, 100%, 100%, and 81.5%, respectively. Because Tzanck smears were evaluated by the same dermatologist, no comment could be made regarding the interobserver reliability of this test and how the level of experience with this technique might affect the results. Also, the sensitivity and the specificity of Tzanck smear test findings for certain diseases could not be calculated because of an insufficient number of patients. The Tzanck smear test is an inexpensive, useful, and an easy diagnostic tool for certain skin diseases.

Comparison of two preparatory techniques for urine cytology.

PubMed Central

Dhundee, J; Rigby, H S

1990-01-01

Two methods of preparation of urine for cytology were compared retrospectively. In method 1 cells in the urine were fixed after the preparation of the smear; in method 2 the cells were fixed before smear preparation. Urine cytology reports were correlated with subsequent histological analysis. The specificities of urine cytology using both methods were high (99%). The sensitivity using method 1 was 87%; using method 2 it was 65%. This difference was significant. The cell preparation technique therefore significantly changes the sensitivity of urine cytology. Cellular fixation after smear preparation is preferable to smear preparation after fixation. PMID:2266176

Under-reporting of sputum smear-positive tuberculosis cases in Kenya.

PubMed

Tollefson, D; Ngari, F; Mwakala, M; Gethi, D; Kipruto, H; Cain, K; Bloss, E

2016-10-01

Although an estimated three million tuberculosis (TB) cases worldwide are missed by national TB programs annually, the level of under-reporting of diagnosed cases in high TB burden settings is largely unknown. To quantify and describe under-reporting of sputum smear-positive TB cases in Kenya. A national-level retrospective TB inventory study was conducted. All sputum smear-positive TB cases diagnosed by public or private laboratories during 1 April-30 June 2013 were extracted from laboratory registers in 73 randomly sampled subcounties and matched to TB cases in the national TB surveillance system (TIBU). Bivariate and multivariate analyses were conducted. In the subcounties sampled, 715 of 3409 smear-positive TB cases in laboratory registers were not found in TIBU. The estimated level of under-reporting of smear-positive TB cases in Kenya was 20.7% (95%CI 18.4-23.0). Under-reporting was greatest in subcounties with a high TB burden. Unreported cases were more likely to be patients aged ⩾55 years, have scanty smear results, and be diagnosed at large facilities, private facilities, and facilities in high TB burden regions. In Kenya, one fifth of smear-positive TB cases diagnosed during the study period went unreported, suggesting that the true TB burden is higher than reported. TB surveillance in Kenya should be strengthened to ensure all diagnosed TB cases are reported.

Historical data decrease complete blood count reflex blood smear review rates without missing patients with acute leukaemia.

PubMed

Rabizadeh, Esther; Pickholtz, Itay; Barak, Mira; Froom, Paul

2013-08-01

The availability of historical data decreases the rate of blood smear review rates in outpatients, but we are unaware of studies done at referral centres. In the following study, we determined the effect of historical data on the rates of peripheral blood smears over a 3-month period and then the detection rate of patients with acute leukaemia. All results of complete blood counts (CBCs) tested on three ADVIA 120 analyzers at the regional Rabin Medical Centre, Beilinson Campus over a 3-month period were accessed on a computerised laboratory information system. Over a 3-month period, we determined the proportion of total CBC and patients with criteria for a manual differential count and the actual number of peripheral blood smears done. Finally, we determined the proportion of 100 consecutive patients with acute leukaemia detected using our criteria that included limiting reflex testing according to historical data. Over the 3-month period, there were 34,827 tests done in 12,785 patients. Without historical data, our smear rate would have been 24.5%, but with the availability of historical data, the blood smear review rate was 5.6%. The detection rate for cases of acute leukaemia was 100%. We conclude that the availability of previous test results significantly reduces the need for blood smear review without missing any patients with acute leukaemia.

High speed wavefront sensorless aberration correction in digital micromirror based confocal microscopy.

PubMed

Pozzi, P; Wilding, D; Soloviev, O; Verstraete, H; Bliek, L; Vdovin, G; Verhaegen, M

2017-01-23

The quality of fluorescence microscopy images is often impaired by the presence of sample induced optical aberrations. Adaptive optical elements such as deformable mirrors or spatial light modulators can be used to correct aberrations. However, previously reported techniques either require special sample preparation, or time consuming optimization procedures for the correction of static aberrations. This paper reports a technique for optical sectioning fluorescence microscopy capable of correcting dynamic aberrations in any fluorescent sample during the acquisition. This is achieved by implementing adaptive optics in a non conventional confocal microscopy setup, with multiple programmable confocal apertures, in which out of focus light can be separately detected, and used to optimize the correction performance with a sampling frequency an order of magnitude faster than the imaging rate of the system. The paper reports results comparing the correction performances to traditional image optimization algorithms, and demonstrates how the system can compensate for dynamic changes in the aberrations, such as those introduced during a focal stack acquisition though a thick sample.

High-quality ultrastructural preservation using cryofixation for 3D electron microscopy of genetically labeled tissues

PubMed Central

Boassa, Daniela; Hu, Junru; Romoli, Benedetto; Phan, Sebastien; Dulcis, Davide

2018-01-01

Electron microscopy (EM) offers unparalleled power to study cell substructures at the nanoscale. Cryofixation by high-pressure freezing offers optimal morphological preservation, as it captures cellular structures instantaneously in their near-native state. However, the applicability of cryofixation is limited by its incompatibility with diaminobenzidine labeling using genetic EM tags and the high-contrast en bloc staining required for serial block-face scanning electron microscopy (SBEM). In addition, it is challenging to perform correlated light and electron microscopy (CLEM) with cryofixed samples. Consequently, these powerful methods cannot be applied to address questions requiring optimal morphological preservation. Here, we developed an approach that overcomes these limitations; it enables genetically labeled, cryofixed samples to be characterized with SBEM and 3D CLEM. Our approach is broadly applicable, as demonstrated in cultured cells, Drosophila olfactory organ and mouse brain. This optimization exploits the potential of cryofixation, allowing for quality ultrastructural preservation for diverse EM applications. PMID:29749931

Optoelectronic hit/miss transform for screening cervical smear slides

NASA Astrophysics Data System (ADS)

Narayanswamy, R.; Turner, R. M.; McKnight, D. J.; Johnson, K. M.; Sharpe, J. P.

1995-06-01

An optoelectronic morphological processor for detecting regions of interest (abnormal cells) on a cervical smear slide using the hit/miss transform is presented. Computer simulation of the algorithm tested on 184 Pap-smear images provided 95% detection and 5% false alarm. An optoelectronic implementation of the hit/miss transform is presented, along with preliminary experimental results.

Immunocytochemistry associated with oral exfoliative cytology: methodological analysis..

PubMed

da Silva, Alessandra Dutra; Lima, Celina Faig; Maraschin, Bruna Jalfim; Laureano, Natália Koerich; Daroit, Natália Batista; Brochier, Fernanda; Sant'Ana Filho, Manoel; Visioli, Fernanda; Rados, Pantelis Varvaki

2015-04-01

To evaluate different immunocytochemical protocol variations to find the most effective protocol for the analysis of involucrin, epidermal growth factor receptor (EGFR), and E-cadherin antibodies. Exfoliative cytology is a noninvasive method used to monitor and screen for early changes in the oral mucosa of patients exposed to carcinogens such as tobacco and alcohol. It has been postulated that its association with immunocytochemistry may improve the effectiveness of the screening process. Four graduate students from Porto Alegre in southern Brazil had oral smears collected from the border of the tongue using a cytobrush. The following variables were analyzed: cell membrane permeability, antigen retrieval method (microwave oven or water bath), antibody incubation time (overnight or 1 hour), detection system used (Envision or LSAB), and chromogen incubation time (10 seconds or 5 minutes). Best results were obtained with the following combinations: (1) for involucrin: water bath, 1-hour incubation for primary antibody, Envision, and chromogen incubation for 10 seconds; (2)for EGFR: microwave, overnight incubation, LSAB, and chromogen incubation for 5 minutes; and (3) for E-cadherin: water bath, over-night incubation, Envision, and chromogen incubation for 5 minutes. Our findings suggest that each antibody requires a specific immunocytochemical protocol to guarantee optimal results with oral smears.

SU (2) lattice gauge theory simulations on Fermi GPUs

NASA Astrophysics Data System (ADS)

Cardoso, Nuno; Bicudo, Pedro

2011-05-01

In this work we explore the performance of CUDA in quenched lattice SU (2) simulations. CUDA, NVIDIA Compute Unified Device Architecture, is a hardware and software architecture developed by NVIDIA for computing on the GPU. We present an analysis and performance comparison between the GPU and CPU in single and double precision. Analyses with multiple GPUs and two different architectures (G200 and Fermi architectures) are also presented. In order to obtain a high performance, the code must be optimized for the GPU architecture, i.e., an implementation that exploits the memory hierarchy of the CUDA programming model. We produce codes for the Monte Carlo generation of SU (2) lattice gauge configurations, for the mean plaquette, for the Polyakov Loop at finite T and for the Wilson loop. We also present results for the potential using many configurations (50,000) without smearing and almost 2000 configurations with APE smearing. With two Fermi GPUs we have achieved an excellent performance of 200× the speed over one CPU, in single precision, around 110 Gflops/s. We also find that, using the Fermi architecture, double precision computations for the static quark-antiquark potential are not much slower (less than 2× slower) than single precision computations.

Filter paper-assisted cell transfer (FaCT) technique: A novel cell-sampling technique for intraoperative diagnosis of central nervous system tumors.

PubMed

Kawamura, Jumpei; Kamoshida, Shingo; Shimakata, Takaaki; Hayashi, Yurie; Sakamaki, Kuniko; Denda, Tamami; Kawai, Kenji; Kuwao, Sadahito

2017-04-01

Intraoperative diagnosis of central nervous system (CNS) tumors provides critical guidance to surgeons in the determination of surgical resection margins and treatment. The techniques and preparations used for the intraoperative diagnosis of CNS tumors include frozen sectioning and cytologic methods (squash smear and touch imprint). Cytologic specimens, which do not have freezing artifacts, are important as an adjuvant tool to frozen sections. However, if the amount of submitted tissue samples is limited, then it is difficult to prepare both frozen sections and squash smears or touch imprint specimens from a single sample at the same time. Therefore, the objective of this study was to derive cells directly from filter paper on which tumor samples are placed. The authors established the filter paper-assisted cell transfer (FaCT) smear technique, in which tumor cells are transferred onto a glass slide directly from the filter paper sample spot after the biopsy is removed. Cell yields and diagnostic accuracy of the FaCT smears were assessed in 40 CNS tumors. FaCT smears had ample cell numbers and well preserved cell morphology sufficient for cytologic diagnosis, even if the submitted tissues were minimal. The overall diagnostic concordance rates between frozen sections and FaCT smears were 90% and 87.5%, respectively (no significant differences). When combining FaCT smears with frozen sections, the diagnostic concordance rate rose to 92.5%. The current results suggest that the FaCT smear technique is a simple and effective processing method that has significant value for intraoperative diagnosis of CNS tumors. Cancer Cytopathol 2017;125:277-282. © 2016 American Cancer Society. © 2017 American Cancer Society.

Is the phone call the most effective method for recall in cervical cancer screening?--results from a randomised control trial.

PubMed

Abdul Rashid, Rima Marhayu; Mohamed, Majdah; Hamid, Zaleha Abdul; Dahlui, Maznah

2013-01-01

To compare the effectiveness of different methods of recall for repeat Pap smear among women who had normal smears in the previous screening. Prospective randomized controlled study. All community clinics in Klang under the Ministry of Health Malaysia. Women of Klang who attended cervical screening and had a normal Pap smear in the previous year, and were due for a repeat smear were recruited and randomly assigned to four different methods of recall for repeat smear. The recall methods given to the women to remind them for a repeat smear were either by postal letter, registered letter, short message by phone (SMS) or phone call. Number and percentage of women who responded to the recall within 8 weeks after they had received the recall, irrespective whether they had Pap test conducted. Also the numbers of women in each recall method that came for repeat Pap smear. The rates of recall messages reaching the women when using letter, registered letter, SMS and phone calls were 79%, 87%, 66% and 68%, respectively. However, the positive responses to recall by letter, registered letter, phone messages and telephone call were 23.9%, 23.0%, 32.9% and 50.9%, respectively (p<0.05). Furthermore, more women who received recall by phone call had been screened (p<0.05) compared to those who received recall by postal letter (OR=2.38, CI=1.56-3.62). Both the usual way of sending letters and registered letters had higher chances of reaching patients compared to using phone either for sending messages or calling. The response to the recall method and uptake of repeat smear, however, were highest via phone call, indicating the importance of direct communication.

Diagnosis of Concurrent Pulmonary Tuberculosis and Tuberculous Otitis Media Confirmed by Xpert MTB/RIF in the United States.

PubMed

Tompkins, Kathleen M; Reimers, Melissa A; White, Becky L; Herce, Michael E

2016-05-01

Tuberculosis (TB) remains an important cause of infectious morbidity in the United States (US), necessitating timely and accurate diagnosis. We report a case of concurrent pulmonary and extrapulmonary TB presenting as tuberculous otitis media in a hospitalized US patient admitted with cough, night sweats, and unilateral purulent otorrhea. Diagnosis was made by smear microscopy and rapidly confirmed by Xpert MTB/RIF-a novel, automated nucleic acid amplification test for the rapid detection of drug-susceptible and drug-resistant TB. This case adds to the growing body of evidence validating Xpert MTB/RIF as an effective tool for the rapid diagnosis of extrapulmonary TB, even in low TB-prevalence settings such as the US, when testing is performed on non-respiratory specimens.

Diagnosis of Concurrent Pulmonary Tuberculosis and Tuberculous Otitis Media Confirmed by Xpert MTB/RIF in the United States

PubMed Central

Tompkins, Kathleen M.; Reimers, Melissa A.; White, Becky L.; Herce, Michael E.

2015-01-01

Tuberculosis (TB) remains an important cause of infectious morbidity in the United States (US), necessitating timely and accurate diagnosis. We report a case of concurrent pulmonary and extrapulmonary TB presenting as tuberculous otitis media in a hospitalized US patient admitted with cough, night sweats, and unilateral purulent otorrhea. Diagnosis was made by smear microscopy and rapidly confirmed by Xpert MTB/RIF—a novel, automated nucleic acid amplification test for the rapid detection of drug-susceptible and drug-resistant TB. This case adds to the growing body of evidence validating Xpert MTB/RIF as an effective tool for the rapid diagnosis of extrapulmonary TB, even in low TB-prevalence settings such as the US, when testing is performed on non-respiratory specimens. PMID:27346926

COMPARISON OF PERMANENT STAINING METHODS FOR THE LABORATORY DIAGNOSIS OF TRICHOMONIASIS

PubMed Central

MENEZES, Camila Braz; MELLO, Mariana dos Santos; TASCA, Tiana

2016-01-01

Trichomonas vaginalis is the etiologic agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in the world. The diagnosis is based on wet mount preparation and direct microscopy on fixed and stained clinical specimens. The aim of this study was to compare the performance of different fixing and staining techniques used in the detection of T. vaginalis in urine. The smears were fixed and submitted to different methods of permanent staining and then, the morphological aspects of the parasites were analyzed and compared. The Papanicolaou staining with ethanol as the fixative solution showed to be the best method of permanent staining. Our data suggest that staining techniques in association with wet mount examination of fresh specimens contribute to increase the sensitivity in the diagnosis of trichomoniasis. PMID:26910452

PAPNET-assisted primary screening of conventional cervical smears.

PubMed

Cenci, M; Nagar, C; Vecchione, A

2000-01-01

The PAPNET System is the only device with a neural-network-based-artificial intelligence to detect and show the images of abnormal cells on the monitor to be evaluated in an interactive way. We effectively used the PAPNET in rescreening of conventional cervical smears and we detected its advantages and its disadvantages. In this paper, we report our results from PAPNET-assisted primary screening performed on 20,154 conventional smears. The smears were classified as Negative or as Review. The Negative cases were rapidly rescreened mainly near the coverslip edges, which are the slide areas not analyzed by automated devices because of focusing problems. The Review cases were fully reanalyzed by the optic microscope. In summary, 140 positive smears were detected: 57 cases showed changes due to HPV, 63 LSIL, 15 HSIL, and 5 carcinomas. Therefore, the PAPNET System was confirmed as useful in primary screening of conventional cervical samples as well as rescreening.

[Clinical application of testing methods on acid-fast bacteria].

PubMed

Ichiyama, Satoshi; Suzuki, Katsuhiro

2005-02-01

Clinical bacteriology pertaining to acid-fast bacteria has made marked advances over the past decade, initiated by the development of a DNA probe kit for identification of acid-fast bacteria. Wide-spread use of nucleic acid amplification for rapid detection of tubercle bacillus contributed more greatly than any other factor to such advances in this field. At present, 90% of all kits used for nucleic acid amplification in the world are consumed in Japan. Unfortunately, not a few clinicians in Japan have a false idea that the smear method and nucleic acid amplification are necessary but culture is not. In any event nucleic acid amplification has exerted significant impacts on the routine works at bacteriology laboratories. Among others, collecting bacteria by pretreatment with NALC-NaOH has simplified the introduction of the collective mode smear method and liquid media. Furthermore, as clinicians have become increasingly more experienced with various methods of molecular biology, it now seems possible to apply these techniques for detection of genes encoding drug resistance and for utilization of molecular epidemiology in routine laboratory works. Meanwhile, attempts to diagnose acid-fast bacteriosis by checking blood for antibody have also been made, primarily in Japan. At present, two kits for detecting antibodies to glycolipids (LAM, TDM, etc.) are covered by national health insurance in Japan. We have an impression that in Japan clinicians do not have adequate knowledge and skill to make full use of these new testing methods clinically. We, as the chairmen of this symposium, hope that this symposium will help clinicians increase their skill related to new testing methods, eventually leading to stimulation of advances in clinical practices related to acid-fast bacteria in Japan. 1. Smear microscopy by concentration method and broth culture system: Kazunari TSUYUGUCHI (Clinical Research Center, National Hospital Organization Kinki-chuo Chest Medical Center) Smear microscopy and culture still remain the cornerstone to diagnose tuberculosis. However, the classical methods in Japan using direct microscopy and Ogawa solid media were not sufficient for clinical use. In recent years substantial advance has been made in these fields. Concentration of clinical samples by centrifugation improves the sensitivity of smear microscopy with excellent reproducibility. The Mycobacteria Growth Indicator Tube (MGIT) system using liquid media yields high sensitivity and rapidity. Using these methods, more and more tuberculosis cases would be correctly diagnosed and treated adequately based on drug susceptibility testing. 2. New technologies for anti-tuberculosis drug susceptibility testing: Satoshi MITARAI (Bacteriology Division, Reference Centre for Mycobacterium, Research Institute of Tuberculosis, Japan Anti-Tuberculosis Association) Several new technologies have been developed to obtain anti-tuberculosis drug susceptibility testing (AST) results rapidly, utilising liquid culture and molecular technologies. Mycobacterium Growth Indicator Tube (MGIT), as a popular liquid culturing and AST system, was evaluated for its accuracy and usefulness. As for isoniazid, MGIT showed 12.6% of discordant result comparing with standard method. These MGIT resistant and Ogawa susceptible strains had relatively high MICs ranging 0.13 to 2.0 microg/ml. The molecular detection of resistant gene mutation is also a useful method to estimate drug resistance rapidly. The rpoB mutation detection is reliable with high sensitivity and specificity. 3. Nucleic acid amplification and novel diagnostic methods: Shunji TAKAKURA (Department of Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine) Sensitivities of nucleic acid amplification tests (NAATs) for the diagnosis of tuberculosis meet clinical requirement that patients with high-risk of transmission should be identified within a day. Comparison of the performance of various NAATs is difficult because of the difference in sample processing and in samples tested among methods and reports. Considering the limitations of NAATs (low sensitivity compared with culture, inability to differentiate dead bacilli from the living), further advances would be expected when novel technologies could confer additional information, such as drug susceptibility, quantity, viability, and genotype. 4. Serodiagnosis of Mycobacterium avium complex lung disease: Seigo KITADA (Department of Internal Medicine, National Hospital Organization Toneyama National Hospital) Mycobacterium avium complex (MAC) organisms are ubiquitous in environment and a contamination in respiratory tracts is sometimes observed, and that complex the diagnosis. We developed a serodiagnostic method for MAC disease using an enzyme immunoassay with the MAC-specific glycopeptidolipid (GPL) core as antigen. A significant increase in GPL core antibodies was detected in sera of patients with MAC pulmonary diseases compared to patients who were colonized with MAC, patients with M. kansasii disease and tuberculosis and healthy subjects. The serodiagnosis is useful for diagnosis of MAC lung disease. 5. Molecular epidemiologic tools for tuberculosis: IS6110 RFLP, Spoligotyping, and VNTR: Tomoshige MATSUMOTO, Hiromi ANO, Tetsuya TAKASHIMA, Izuo TSUYUGUCHI (Osaka Prefectural Medical Center for Respiratory and Allergic Diseases) We have performed molecular typing on about 1,300 culture positive clinical isolates that made up the majority of tuberculosis strains in part of southeast Osaka since 2001 until now. By spoligotyping, about 75% of entire strains belonged to the Beijing strain. Particular spoligotyping descriptions, which were not described in SpolDBIII, were found in the strains with lower than 6 copies of IS6110 RFLP. We described them as Osaka type. We could also show that direct typing from Tb PCR positive sputum of patients with tuberculosis was possible by VNTR and that VNTR with 16 loci was useful in tuberculosis typing in Osaka.

The change is length and width of the Sertoli cell nuclei in cytologic smears of testes with depopulation of the seminiferous epithelium.

PubMed

Banek, L; Posinovec, J

1980-09-15

The appearance of the Sertoli cells in cytological smears of tests with depopulation of the seminiferous epithelium is described. The mean values of the lengths and widths of the Sertoli cell nuclei in smears differed significantly between the depopulation and the control group (p < 0.01).

Papanicolaou Smear Screening of Women with Intellectual Disabilities: A Cross-Sectional Survey in Taiwan

ERIC Educational Resources Information Center

Lin, Lan-Ping; Lin, Jin-Ding; Sung, Chang-Lin; Liu, Ta-Wen; Liu, Yi-Lian; Chen, Li-Mei; Chu, Cordia M.

2010-01-01

Although little is known about the incidence of cervical cancer in women with intellectual disabilities (ID), Pap smear screening is an effective public health program to prevent cervical cancer to this group of people. The purposes of this study were to identify and evaluate the factors regarding the utilization of the Pap smears in women with ID…

Natural History of Tuberculosis: Duration and Fatality of Untreated Pulmonary Tuberculosis in HIV Negative Patients: A Systematic Review

PubMed Central

Tiemersma, Edine W.; van der Werf, Marieke J.; Borgdorff, Martien W.; Williams, Brian G.; Nagelkerke, Nico J. D.

2011-01-01

Background The prognosis, specifically the case fatality and duration, of untreated tuberculosis is important as many patients are not correctly diagnosed and therefore receive inadequate or no treatment. Furthermore, duration and case fatality of tuberculosis are key parameters in interpreting epidemiological data. Methodology and Principal Findings To estimate the duration and case fatality of untreated pulmonary tuberculosis in HIV negative patients we reviewed studies from the pre-chemotherapy era. Untreated smear-positive tuberculosis among HIV negative individuals has a 10-year case fatality variously reported between 53% and 86%, with a weighted mean of 70%. Ten-year case fatality of culture-positive smear-negative tuberculosis was nowhere reported directly but can be indirectly estimated to be approximately 20%. The duration of tuberculosis from onset to cure or death is approximately 3 years and appears to be similar for smear-positive and smear-negative tuberculosis. Conclusions Current models of untreated tuberculosis that assume a total duration of 2 years until self-cure or death underestimate the duration of disease by about one year, but their case fatality estimates of 70% for smear-positive and 20% for culture-positive smear-negative tuberculosis appear to be satisfactory. PMID:21483732

Effects of spectral smearing on performance of the spectral ripple and spectro-temporal ripple tests.

PubMed

Narne, Vijaya Kumar; Sharma, Mridula; Van Dun, Bram; Bansal, Shalini; Prabhu, Latika; Moore, Brian C J

2016-12-01

The main aim of this study was to use spectral smearing to evaluate the efficacy of a spectral ripple test (SRt) using stationary sounds and a recent variant with gliding ripples called the spectro-temporal ripple test (STRt) in measuring reduced spectral resolution. In experiment 1 the highest detectable ripple density was measured using four amounts of spectral smearing (unsmeared, mild, moderate, and severe). The thresholds worsened with increasing smearing and were similar for the SRt and the STRt across the three conditions with smearing. For unsmeared stimuli, thresholds were significantly higher (better) for the STRt than for the SRt. An amplitude fluctuation at the outputs of simulated (gammatone) auditory filters centered above 6400 Hz was identified as providing a potential detection cue for the STRt stimuli. Experiment 2 used notched noise with energy below and above the passband of the SRt and STRt stimuli to reduce confounding cues in the STRt. Thresholds were almost identical for the STRt and SRt for both unsmeared and smeared stimuli, indicating that the confounding cue for the STRt was eliminated by the notched noise. Thresholds obtained with notched noise present could be predicted reasonably accurately using an excitation-pattern model.

Cervical cancer screening among Lebanese women.

PubMed

Bou-Orm, I R; Sakr, R E; Adib, S M

2018-02-01

Cervical cancer is a very common malignancy amongst women worldwide. Pap smear is an effective and inexpensive screening test in asymptomatic women. The aim of this paper was to assess the prevalence of Pap smear screening for cervical cancer among Lebanese women and to determine associated sociodemographic and psychosocial characteristics. This national survey included 2255 women, selected by multi-stage random cluster sampling across Lebanon. A questionnaire about practices and perceptions related to cervical cancer screening was developed based on the "Health Belief Model". The weighted national prevalence of "ever-use" of the Pap smear for screening purposes was 35%. Most important determinants of screening behavior were: residence within Greater Beirut, higher socio-economic status and educational attainment, marriage status, presence of a health coverage, awareness of Pap smear usefulness, knowing someone who had already done it, and a balance between perceived benefits and perceived barriers to Pap smear screening. Regular information campaigns regarding the availability and effectiveness of the test should be devised, targeting in priority the sexually vulnerable women in Lebanon. Moreover, healthcare providers should be encouraged to discuss with their patients the opportunity of obtaining a Pap smear. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Predicting radiative heat transfer in thermochemical nonequilibrium flow fields. Theory and user's manual for the LORAN code

NASA Technical Reports Server (NTRS)

Chambers, Lin Hartung

1994-01-01

The theory for radiation emission, absorption, and transfer in a thermochemical nonequilibrium flow is presented. The expressions developed reduce correctly to the limit at equilibrium. To implement the theory in a practical computer code, some approximations are used, particularly the smearing of molecular radiation. Details of these approximations are presented and helpful information is included concerning the use of the computer code. This user's manual should benefit both occasional users of the Langley Optimized Radiative Nonequilibrium (LORAN) code and those who wish to use it to experiment with improved models or properties.

Cancer of the cervix - from bleak past to bright future; a review, with an emphasis on cancer of the cervix in malaysia.

PubMed

Nor Hayati, Othman

2003-01-01

Cancer of the cervix has the potential to be eradicated since the initiating cause is known. There was not much known about this cancer until the time of the Renaissance. In Malaysia, it is the second most common cancer among females after breast cancer. The strategies on prevention in this country are still not optimal. This article highlights the problems and also discusses the pathogenesis of this disease. The key to prevention is screening and the future is the era of molecular pap smear.

Association between Pap smear screening and job stress in Taiwanese nurses.

PubMed

Su, Shiang-Yuan; Chiou, Shu-Ti; Huang, Nicole; Huang, Chiu-Mieh; Chiang, Jen-Huai; Chien, Li-Yin

2016-02-01

Nursing is a professional job characterized by high stress. Stress could be associated with less practice of health promoting behaviors; however, no study has investigated the relationship between job stress and health screening behaviors among nurses. This study aimed to describe the rate of Pap smears in hospital nurses and examine the effects of job stress on receiving a Pap smear. This study was a cross-sectional survey. The study participants were 30,681 full-time female nurses who were at least 30 years of age working in 100 hospitals across Taiwan. The study participants filled out an anonymous structured questionnaire from May to July, 2011. The outcome variable was having a Pap test during the previous 3 years. The level of stress was measured by a 19-item scale, with higher scores indicating higher stress levels. About 62.4% of the nurses had a Pap smear during the previous three years. Each point increase in the stress score decreased the likelihood of Pap smears (OR = .997, 95% CI: .995-.999), after adjustment for participant characteristics, health status, health behaviors, and hospital characteristics. Despite more knowledge and higher accessibility, nurses were less likely to have Pap smear screening than the general population. A higher level of job stress was associated with a lower likelihood of having a Pap smear. Hospital administrators could help decrease work-related stress and improve stress adaption among nurses in order to improve their health screening behaviors. Copyright © 2015 Elsevier Ltd. All rights reserved.

Comparative analysis of endodontic smear layer removal efficacy of 17% ethylenediaminetetraacetic acid, 7% maleic acid, and 2% chlorhexidine using scanning electron microscope: An in vitro study.

PubMed

Attur, Kailash; Joy, Mathew T; Karim, Riyas; Anil Kumar, V J; Deepika, C; Ahmed, Haseena

2016-08-01

The aim of the present study was to evaluate the efficiency of different endodontic irrigants in the removal of smear layer through scanning electron microscopic image analysis. The present in vitro study was carried out on 45 single-rooted extracted human mandibular premolar teeth with single canal and complete root formation. Teeth were randomly assigned to three groups with 15 teeth in each group. Group I samples were irrigated with 17% ethylenediaminetetraacetic (EDTA) irrigation, Group II with 7% maleic acid irrigation, and Group III with 2% chlorhexidine irrigation. Scanning electron microscope evaluation was done for the assessment of smear layer removal in the coronal, middle, and apical thirds. Comparison of the smear layer removal between the three different groups was done by Kruskal-Wallis test, followed by Mann-Whitney U test for comparing individual groups. A P value less than 0.05 was considered to be statistically significant. Statistically significant difference was seen between the two test groups (17% EDTA vs. 7% maleic acid and 17% EDTA vs. 2% chlorhexidine) in smear layer removal at coronal, middle, and apical thirds of the root canal. The most efficient smear layer removal was seen in Group I with 17% EDTA irrigation compared with other groups (P < 0.05) and the least by 2% chlorhexidine. The present study shows that 17% EDTA efficiently removes the smear layer from root canal walls.

Effects of overnight refrigeration on the microscopic evaluation of sputum.

PubMed Central

Penn, R L; Silberman, R

1984-01-01

Microscopic evaluation of sputum permits selection of specimens suitable for culture, assessment of likely pathogens, and the best interpretation of culture results. We prospectively evaluated 50 sputum specimens which were promptly submitted to our clinical laboratory; smears and cultures were performed both immediately and after 20 h of refrigeration. Specimens were grouped according to the numbers of squamous epithelial cells and neutrophils per low-power field present on coded Gram-stained smears. The numbers of bacteria in five oil immersion fields were used to characterize smears for predominant, mixed, or scanty forms. After refrigeration, only three specimens changed group from a definite loss of squamous epithelial cells, and only two changed group from a definite loss of neutrophils. Based on cellular composition, the majority of samples would have been processed identically both before and after refrigeration. In contrast, organism forms detected on smears and their relative quantities were dramatically altered after refrigeration. A predominant smear form was gained in 11 and lost in 8 refrigerated specimens. The frequent changes on smears observed overall resulted from both increases and decreases in numbers of bacteria and yeasts. The majority of sputum culture results were insignificantly affected by the refrigeration of specimens. We conclude that 20 h of refrigeration renders sputum useless for the microscopic evaluation of potential pathogens and the subsequent interpretation of culture results. However, overnight refrigeration does not affect the determination from smears of sputum suitability for culture based on cellular composition. PMID:6699145

Stochastic, compartmental, and dynamic modeling of cross-contamination during mechanical smearing of cheeses.

PubMed

Aziza, Fanny; Mettler, Eric; Daudin, Jean-Jacques; Sanaa, Moez

2006-06-01

Cheese smearing is a complex process and the potential for cross-contamination with pathogenic or undesirable microorganisms is critical. During ripening, cheeses are salted and washed with brine to develop flavor and remove molds that could develop on the surfaces. Considering the potential for cross-contamination of this process in quantitative risk assessments could contribute to a better understanding of this phenomenon and, eventually, improve its control. The purpose of this article is to model the cross-contamination of smear-ripened cheeses due to the smearing operation under industrial conditions. A compartmental, dynamic, and stochastic model is proposed for mechanical brush smearing. This model has been developed to describe the exchange of microorganisms between compartments. Based on the analytical solution of the model equations and on experimental data collected with an industrial smearing machine, we assessed the values of the transfer parameters of the model. Monte Carlo simulations, using the distributions of transfer parameters, provide the final number of contaminated products in a batch and their final level of contamination for a given scenario taking into account the initial number of contaminated cheeses of the batch and their contaminant load. Based on analytical results, the model provides indicators for smearing efficiency and propensity of the process for cross-contamination. Unlike traditional approaches in mechanistic models, our approach captures the variability and uncertainty inherent in the process and the experimental data. More generally, this model could represent a generic base to use in modeling similar processes prone to cross-contamination.

A comparative evaluation of smear layer removal by using edta, etidronic acid, and maleic acid as root canal irrigants: An in vitro scanning electron microscopic study

PubMed Central

Kuruvilla, Aby; Jaganath, Bharath Makonahalli; Krishnegowda, Sahadev Chickmagaravalli; Ramachandra, Praveen Kumar Makonahalli; Johns, Dexton Antony; Abraham, Aby

2015-01-01

Aim: The purpose of this study is to evaluate and compare the efficacy of 17% EDTA, 18% etidronic acid, and 7% maleic acid in smear layer removal using scanning electron microscopic image analysis. Materials and Methods: Thirty, freshly extracted mandibular premolars were used. The teeth were decoronated to obtain working length of 17mm and instrumentation up to 40 size (K file) with 2.5% NaOCl irrigation between each file. The samples were divided into Groups I (17% ethylenediaminetetraacetic acid (EDTA)), II (18% etidronic acid), and III (7% maleic acid) containing 10 samples each. Longitudinal sectioning of the samples was done. Then the samples were observed under scanning electron microscope (SEM) at apical, middle, and coronal levels. The images were scored according to the criteria: 1. No smear layer, 2. moderate smear layer, and 3 heavy smear layer. Statistical Analysis: Data was analyzed statistically using Kruskal–Wallis analysis of variance (ANOVA) followed by Mann-Whitney U test for individual comparisons. The level for significance was set at 0.05. Results: The present study showed that all the three experimental irrigants removed the smear layer from different tooth levels (coronal, middle, and apical). Final irrigation with 7% maleic acid is more efficient than 17% EDTA and 18% etidronic acid in the removal of smear layer from the apical third of root canal. PMID:26069414

Laboratory productivity and the rate of manual peripheral blood smear review: a College of American Pathologists Q-Probes study of 95,141 complete blood count determinations performed in 263 institutions.

PubMed

Novis, David A; Walsh, Molly; Wilkinson, David; St Louis, Mary; Ben-Ezra, Jonathon

2006-05-01

Automated laboratory hematology analyzers are capable of performing differential counts on peripheral blood smears with greater precision and more accurate detection of distributional and morphologic abnormalities than those performed by manual examinations of blood smears. Manual determinations of blood morphology and leukocyte differential counts are time-consuming, expensive, and may not always be necessary. The frequency with which hematology laboratory workers perform manual screens despite the availability of labor-saving features of automated analyzers is unknown. To determine the normative rates with which manual peripheral blood smears were performed in clinical laboratories, to examine laboratory practices associated with higher or lower manual review rates, and to measure the effects of manual smear review on the efficiency of generating complete blood count (CBC) determinations. From each of 3 traditional shifts per day, participants were asked to select serially, 10 automated CBC specimens, and to indicate whether manual scans and/or reviews with complete differential counts were performed on blood smears prepared from those specimens. Sampling continued until a total of 60 peripheral smears were reviewed manually. For each specimen on which a manual review was performed, participants indicated the patient's age, hemoglobin value, white blood cell count, platelet count, and the primary reason why the manual review was performed. Participants also submitted data concerning their institutions' demographic profiles and their laboratories' staffing, work volume, and practices regarding CBC determinations. The rates of manual reviews and estimations of efficiency in performing CBC determinations were obtained from the data. A total of 263 hospitals and independent laboratories, predominantly located in the United States, participating in the College of American Pathologists Q-Probes Program. There were 95,141 CBC determinations examined in this study; participants reviewed 15,423 (16.2%) peripheral blood smears manually. In the median institution (50th percentile), manual reviews of peripheral smears were performed on 26.7% of specimens. Manual differential count review rates were inversely associated with the magnitude of platelet counts that were required by laboratory policy to trigger smear reviews and with the efficiency of generating CBC reports. Lower manual differential count review rates were associated with laboratory policies that allowed manual reviews solely on the basis of abnormal automated red cell parameters and that precluded performing repeat manual reviews within designated time intervals. The manual scan rate elevated with increased number of hospital beds. In more than one third (35.7%) of the peripheral smears reviewed manually, participants claimed to have learned additional information beyond what was available on automated hematology analyzer printouts alone. By adopting certain laboratory practices, it may be possible to reduce the rates of manual reviews of peripheral blood smears and increase the efficiency of generating CBC results.

Hepatozoon caimani Carini, 1909 (Adeleina: Hepatozoidae) in wild population of Caiman yacare Daudin, 1801 (Crocodylia: Alligatoridae), Pantanal, Brazil.

PubMed

Soares, Priscilla; Borghesan, Tarcilla Corrente; Tavares, Luiz Eduardo Rolland; Ferreira, Vanda Lúcia; Teixeira, Marta Maria Geraldes; Paiva, Fernando

2017-07-01

Previous studies showed infections of Hepatozoon caimani in wild populations of caimans in wide regions from Brazil; some of those demonstrated that trophic chain are linked to natural infections through paratenic hosts or by the direct ingestion of vectors. These studies life cycle of H. caimani contributed inestimably to the knowledge of transmission routes, yet but lack enhancement tools for better detail of parasite. This study reports the forms in the blood and tissues, and also partial molecular characterization of the H. caimani following part of the 18S rRNA region. In the southern Pantanal, there were sampling 39 adult caimans (Caiman yacare), where 31 (79.5%) were parasitized by H. caimani. Free gametocytes had an average intensity of 19.6% and intraerythrocytic forms 7.42%, in the blood smears. In stained smears of the liver and lungs of naturally infected caimans which were examined, monozoic and dizoic cysts were found in these tissues, generally next to the vessels. In the histopathology, meronts were observed in the wall of vessels from liver and kidney ducts. Blood samples were forwarded to PCR process and produced amplicons with about 600 and 900 bp, respectively, for the primers HEPF300/HEP900 and HEMO1/HEMO2. This was the first report of molecular confirmation of Hepatozoon in populations of naturally infected caimans of morphological detail of the gametocytes in scanning electron microscopy and histology of merogony in livers and kidneys of C. yacare.

Automated system for characterization and classification of malaria-infected stages using light microscopic images of thin blood smears.

PubMed

Das, D K; Maiti, A K; Chakraborty, C

2015-03-01

In this paper, we propose a comprehensive image characterization cum classification framework for malaria-infected stage detection using microscopic images of thin blood smears. The methodology mainly includes microscopic imaging of Leishman stained blood slides, noise reduction and illumination correction, erythrocyte segmentation, feature selection followed by machine classification. Amongst three-image segmentation algorithms (namely, rule-based, Chan-Vese-based and marker-controlled watershed methods), marker-controlled watershed technique provides better boundary detection of erythrocytes specially in overlapping situations. Microscopic features at intensity, texture and morphology levels are extracted to discriminate infected and noninfected erythrocytes. In order to achieve subgroup of potential features, feature selection techniques, namely, F-statistic and information gain criteria are considered here for ranking. Finally, five different classifiers, namely, Naive Bayes, multilayer perceptron neural network, logistic regression, classification and regression tree (CART), RBF neural network have been trained and tested by 888 erythrocytes (infected and noninfected) for each features' subset. Performance evaluation of the proposed methodology shows that multilayer perceptron network provides higher accuracy for malaria-infected erythrocytes recognition and infected stage classification. Results show that top 90 features ranked by F-statistic (specificity: 98.64%, sensitivity: 100%, PPV: 99.73% and overall accuracy: 96.84%) and top 60 features ranked by information gain provides better results (specificity: 97.29%, sensitivity: 100%, PPV: 99.46% and overall accuracy: 96.73%) for malaria-infected stage classification. © 2014 The Authors Journal of Microscopy © 2014 Royal Microscopical Society.

Definition of a type of abnormal vaginal flora that is distinct from bacterial vaginosis: aerobic vaginitis.

PubMed

Donders, Gilbert G G; Vereecken, Annie; Bosmans, Eugene; Dekeersmaecker, Alfons; Salembier, Geert; Spitz, Bernard

2002-01-01

To define an entity of abnormal vaginal flora: aerobic vaginitis. Observational study. University Hospital Gasthuisberg, Leuven, Belgium. 631 women attending for routine prenatal care or attending vaginitis clinic. Samples were taken for fresh wet mount microscopy of vaginal fluid, vaginal cultures and measurement of lactate, succinate and cytokine levels in vaginal fluid. Smears deficient in lactobacilli and positive for clue cells were considered to indicate a diagnosis of bacterial vaginosis. Aerobic vaginitis was diagnosed if smears were deficient in lactobacilli, positive for cocci or coarse bacilli, positive for parabasal epithelial cells, and/or positive for vaginal leucocytes (plus their granular aspect). Genital complaints include red inflammation, yellow discharge, vaginal dyspareunia. Group B streptococci, escherichia coli, staphylococcus aureus and trichomonas vaginalis are frequently cultured. Vaginal lactate concentration is severely depressed in women with aerobic vaginitis, as in bacterial vaginosis, but vaginal succinate is not produced. Also in contrast to bacterial vaginosis, aerobic vaginitis produces a host immune response that leads to high production of interleukin-6, interleukin-1-beta and leukaemia inhibitory factor in the vaginal fluid. Aerobic vaginitis is associated with aerobic micro-organisms, mainly group B streptococci and E. coli. Its characteristics are different from those of bacterial vaginosis and elicit an important host response. The most severe form of aerobic vaginitis equals desquamative inflammatory vaginitis. In theory, aerobic vaginitis may be a better candidate than bacterial vaginosis as the cause of pregnancy complications, such as ascending chorioamnionitis, preterm rupture of the membranes and preterm delivery.

In vivo optical detection of intranuclear cancer biomarkers using gold nanoparticles

NASA Astrophysics Data System (ADS)

Kumar, Sonia; Sokolov, Konstantin; Richards-Kortum, Rebecca

2006-02-01

Specific genotypes of human papillomavirus (HPV) are well correlated with cervical oncogenesis. The major transforming and immortalizing protein in high risk HPVs, namely HPV16, is E7 protein. E7 protein functions by deregulating the cell cycle and promoting S-phase reentry in differentiated keratinocytes. Currently, clinical diagnosis of cervical cancer is based on phenotypic changes observed in a screening Papanicolaou smear. Although screening has been effective in reducing the occurrence of cervical cancer, the low specificity of the Pap smear results in resources wasted on the evaluation of low-grade lesions not likely to progress to cervical cancer. Molecular characterization of active HPV infections using molecular specific contrast agents are combined with in-vivo optical imaging is proposed to be a cost-effective, non-invasive technique for the detection of cervical pre-cancers. Contrast is achieved by exploiting the peak absorbance and scattering shift in aggregated gold nanoparticles over isolated ones and molecular specificity is achieved via recognition moieties with high affinities for E7. Conjugates of gold nanoparticles and HPV16 anti-E7 antibodies are delivered into the nucleus of living cells and imaged with reflectance confocal microscopy. These contrast agents have been used to successfully enhance contrast in HPV16+ cervical cancer cells over HPV- cells by a factor of 2.5. Further characterization and development of these contrast agents will provide a robust, low cost screening tool for the detection of cervical pre-cancers.

Contact Investigation in Households of Patients with Tuberculosis in Hanoi, Vietnam: A Prospective Cohort Study

PubMed Central

Fox, Gregory James; Nhung, Nguyen Viet; Sy, Dinh Ngoc; Lien, Luu Thi; Cuong, Nguyen Kim; Britton, Warwick John; Marks, Guy Barrington

2012-01-01

Setting Existing tuberculosis control strategies in Vietnam are based on symptomatic patients attending health services for investigation. This approach has not resulted in substantial reductions in the prevalence of tuberculosis disease, despite the National Tuberculosis Program achieving high treatment completion rates. Alternative approaches are being considered. Objective To determine the feasibility and yield of contact investigation in households of patients with smear positive pulmonary tuberculosis among household members of tuberculosis patients in Hanoi, Vietnam. Methods Household contacts of patients with smear positive pulmonary tuberculosis were recruited at four urban and rural District Tuberculosis Units in Hanoi. Clinical and radiological screening was conducted at baseline, six months and 12 months. Sputum microscopy and culture was performed in contacts suspected of having tuberculosis. MIRU-VNTR molecular testing was used to compare the strains of patients and their contacts with disease. Results Among 545 household contacts of 212 patients, four were diagnosed with tuberculosis at baseline (prevalence 734 cases per 100,000 persons, 95% CI 17–1451) and one was diagnosed with tuberculosis during the subsequent 12 months after initial screening (incidence 180 cases per 100,000 person-years, 95% CI 44–131). Two of these cases were culture positive for M. tuberculosis and both had identical or near-identical MIRU-VNTR strain types. Conclusion Household contacts of patients with potentially infectious forms of tuberculosis have a high prevalence of disease. Household contact investigation is feasible in Vietnam. Further research is required to investigate its effectiveness. PMID:23166785

Pediatric tuberculosis-human immunodeficiency virus co-infection in the United Kingdom highlights the need for better therapy monitoring tools: a case report.

PubMed

Evangelopoulos, Dimitrios; Whittaker, Elizabeth; Honeyborne, Isobella; McHugh, Timothy D; Klein, Nigel; Shingadia, Delane

2017-02-26

Tuberculosis is an infection that requires at least 6 months of chemotherapy in order to clear the bacteria from the patient's lungs. Usually, therapeutic monitoring is dependent on smear microscopy where a decline in acid-fast bacilli is observed. However, this might not be indicative of the actual decline of bacterial load and thus other tools such as culture and molecular assays are required for patient management. Here, we report the case of a 12-year-old Black African boy co-infected with tuberculosis and human immunodeficiency virus who remained smear culture positive and liquid culture negative for a prolonged period of time following chemotherapy. In order to determine whether there was any live bacteria present in his specimens, we applied the newly developed molecular bacterial load assay that detects the presence of 16S ribosomal ribonucleic acid derived from the bacteria. Using this methodology, we were able to quantify his bacterial load and inform the management of his treatment in order to reduce the disease burden. Following this intervention he went on to make a complete recovery. This case report highlights the value of improved biomarkers for monitoring the treatment of tuberculosis and the role of molecular assays such as the molecular bacterial load assay applied here. The molecular bacterial load assay detects bacterial ribonucleic acid which corresponds closely with the number of live bacilli as compared with polymerase chain reaction that detects deoxyribonucleic acid and may include dead bacteria.

Smear Layer Removal in the Apical Third of Root Canals by Two Chelating Agents and Laser: A Comparative in vitro Study

PubMed Central

Ashraf, Hengameh; Asnaashari, Mohammad; Darmiani, Soheila; Birang, Reza

2014-01-01

Introduction: Smear layer (SL) is produced as a result of mechanical instrumentation of the canal(s). Despite the controversies regarding its removal, the evidence-based trend has shifted towards removing and eliminating the SL. Different methods have been used to remove the SL and the aim of this in vitro study was to evaluate the ability of 17% ethylenediaminetetraacetic acid (EDTA), 18% etidronate and Er: YAG on effective removal of the SL. Methods and Materials: Fifty straight single-rooted teeth were divided into three experimental groups (n=15) and one control group of five. The canals were instrumented with HERO 642 rotary files up to 30/0.06. In group 1, canals were irradiated with Er: YAG laser; in groups 2 and 3, canals were irrigated with 17% EDTA and 18% etidronate, respectively. In group 4 (control) distilled water was used for canal irrigation. The amount of remaining SL was quantified according to Hulsmann’s method with scanning electron microscopy (SEM). Data was analyzed by the Kruskal-Wallis and Mann-Whitney U tests (P<0.05). Results: The results showed statistically significant differences in terms of SL removal among the groups (P<0.05). The amount of removed SL by EDTA was significantly greater followed by Er: YAG laser and 18% etidronate. Conclusion: Within the limitations of this study, EDTA was more effective in removing SL compared to Er: YAG and etidronate. PMID:25031596

Imaging multicellular specimens with real-time optimized tiling light-sheet selective plane illumination microscopy

PubMed Central

Fu, Qinyi; Martin, Benjamin L.; Matus, David Q.; Gao, Liang

2016-01-01

Despite the progress made in selective plane illumination microscopy, high-resolution 3D live imaging of multicellular specimens remains challenging. Tiling light-sheet selective plane illumination microscopy (TLS-SPIM) with real-time light-sheet optimization was developed to respond to the challenge. It improves the 3D imaging ability of SPIM in resolving complex structures and optimizes SPIM live imaging performance by using a real-time adjustable tiling light sheet and creating a flexible compromise between spatial and temporal resolution. We demonstrate the 3D live imaging ability of TLS-SPIM by imaging cellular and subcellular behaviours in live C. elegans and zebrafish embryos, and show how TLS-SPIM can facilitate cell biology research in multicellular specimens by studying left-right symmetry breaking behaviour of C. elegans embryos. PMID:27004937

Effect of diabetes on tuberculosis presentation and outcomes in Kiribati.

PubMed

Cavanaugh, J; Viney, K; Kienene, T; Harley, D; Kelly, P M; Sleigh, A; O'Connor, J; Mase, S

2015-05-01

To determine the association between diabetes and the clinical features and treatment outcomes of TB in Kiribati. We enrolled consecutive patients with TB who presented from August 2010 to February 2012 and compared clinical features and TB treatment outcomes for patients with and without diabetes, as measured by haemoglobin A1c assay. Poor outcome was defined as death, default or treatment failure, and good outcome as treatment success or cure. Two hundred and seventy-five eligible persons with TB disease were enrolled; 101 (37%) had diabetes. TB patients with diabetes were more likely to have acid-fast bacilli (AFB) seen on sputum smear microscopy (RR: 1.3; 95% CI: 1.03-1.62). The risk of poor outcome did not differ between patients with or without diabetes (RR: 1.1; 95% CI: 0.5-2.7). TB patients with diabetes are more likely than those without to have sputum with AFB on microscopy. This could increase transmission in the community. Early detection of TB by screening patients with diabetes, and the converse, could be important public health interventions where diabetes and TB are prevalent. © 2015 John Wiley & Sons Ltd.

Detection of avian malaria (Plasmodium spp.) in native land birds of American Samoa

USGS Publications Warehouse

Jarvi, S.I.; Farias, M.E.M.; Baker, H.; Freifeld, H.B.; Baker, P.E.; Van Gelder, E.; Massey, J.G.; Atkinson, C.T.

2003-01-01

This study documents the presence of Plasmodium spp. in landbirds of central Polynesia. Blood samples collected from eight native and introduced species from the island of Tutuila, American Samoa were evaluated for the presence of Plasmodium spp. by nested rDNA PCR, serology and/or microscopy. A total of 111/188 birds (59%) screened by nested PCR were positive. Detection of Plasmodium spp. was verified by nucleotide sequence comparisons of partial 18S ribosomal RNA and TRAP (thrombospondin-related anonymous protein) genes using phylogenetic analyses. All samples screened by immunoblot to detect antibodies that cross-react with Hawaiian isolates of Plasmodium relictum (153) were negative. Lack of cross-reactivity is probably due to antigenic differences between the Hawaiian and Samoan Plasmodium isolates. Similarly, all samples examined by microscopy (214) were negative. The fact that malaria is present, but not detectable by blood smear evaluation is consistent with low peripheral parasitemia characteristic of chronic infections. High prevalence of apparently chronic infections, the relative stability of the native land bird communities, and the presence of mosquito vectors which are considered endemic and capable of transmitting avian Plasmodia, suggest that these parasites are indigenous to Samoa and have a long coevolutionary history with their hosts.

Plasmodium ovale infection in Malaysia: first imported case

PubMed Central

2010-01-01

Background Plasmodium ovale infection is rarely reported in Malaysia. This is the first imported case of P. ovale infection in Malaysia which was initially misdiagnosed as Plasmodium vivax. Methods Peripheral blood sample was first examined by Giemsa-stained microscopy examination and further confirmed using a patented in-house multiplex PCR followed by sequencing. Results and Discussion Initial results from peripheral blood smear examination diagnosed P. vivax infection. However further analysis using a patented in-house multiplex PCR followed by sequencing confirmed the presence of P. ovale. Given that Anopheles maculatus and Anopheles dirus, vectors of P. ovale are found in Malaysia, this finding has significant implication on Malaysia's public health sector. Conclusions The current finding should serve as an alert to epidemiologists, clinicians and laboratory technicians in the possibility of finding P. ovale in Malaysia. P. ovale should be considered in the differential diagnosis of imported malaria cases in Malaysia due to the exponential increase in the number of visitors from P. ovale endemic regions and the long latent period of P. ovale. It is also timely that conventional diagnosis of malaria via microscopy should be coupled with more advanced molecular tools for effective diagnosis. PMID:20929588

In vivo Susceptibility of Plasmodium vivax to Chloroquine in Southeastern Iran.

PubMed

Heidari, A; Keshavarz, H; Shojaee, S; Raeisi, A; Dittrich, S

2012-01-01

Plasmodium vivax is the predominant species causes of malaria with about 90% total annual reported malaria in Iran. This study conducted to determine the susceptibility of Plasmodium vivax isolates to chloroquine in Sistan and Balochistan Province, southeastern Iran. A total 270 subjects with symptomatic malaria and confirmed P. vivax infection completed the designed 28-day in vivo study. The thick and thin film blood smears were screened for malaria parasites by microscopy. The nested PCR was applied using the Plasmodium 18 subunit ribosomal ribonucleic (Ssr RNA) genes for detecting mixed infections and diagnosis of parasites in the samples with low parasite on days 0, 5, 6, 7, and 28. P. vivax was cleared in 15%, 50%, 95%, and 100% of patients on days 1, 2, 3, 4 respectively by microscopy assessment. Six patients were exhibited specific P. vivax band in nested PCR on day 5. No recurrence was observed on days 7, 14 and 28. Mean (±standard deviation) parasite clearance time was 2.41 (±0.8) days. P. vivax is still susceptible to chloroquine in Southeatern Iran. This finding is compatible with results of neighboring countries Pakistan and Afghanistan.

Malaria in pregnant Cameroonian women: the effect of age and gravidity on submicroscopic and mixed-species infections and multiple parasite genotypes.

PubMed

Walker-Abbey, Annie; Djokam, Rosine R T; Eno, Anna; Leke, Rose F G; Titanji, Vincent P K; Fogako, Josephine; Sama, Grace; Thuita, Lucy H; Beardslee, Eliza; Snounou, Georges; Zhou, Ainong; Taylor, Diane Wallace

2005-03-01

Polymerase chain reaction (PCR)-based methods were used to investigate malaria in pregnant women residing in Yaounde, Cameroon. Microscopy and species-specific PCR-based diagnosis show that at delivery 82.4% of the women were infected with Plasmodium falciparum (27.5% blood-smear positive and 54.9% submicroscopic infections). The prevalence of P. malariae and P. ovale was 7.6% and 2.5%, respectively, with 9.4% infected with more than one species. Based on genotyping of the merozoite surface protein 1 (msp-1) and msp-2 alleles, the mean number of genetically different P. falciparum parasites in peripheral blood was 3.4 (range = 1-9) and 3.5 (range 1-8) in the placenta. Plasmodium falciparum detected by microscopy and PCR as well as mixed-species infections were significantly higher in women < or = 20 years old and paucigravidae, but maternal anemia was associated only with microscopic detection of parasites. Neither submicroscopic infections nor number of parasite genotypes decreased significantly with age or gravidity. Thus, pregnancy-associated immunity helps reduce malaria to submicroscopic levels, but does not reduce the number of circulating parasite genotypes.