Rapid detection of Salmonella spp. in food by use of the ISO-GRID hydrophobic grid membrane filter.

PubMed Central

Entis, P; Brodsky, M H; Sharpe, A N; Jarvis, G A

1982-01-01

A rapid hydrophobic grid-membrane filter (HGMF) method was developed and compared with the Health Protection Branch cultural method for the detection of Salmonella spp. in 798 spiked samples and 265 naturally contaminated samples of food. With the HGMF method, Salmonella spp. were isolated from 618 of the spiked samples and 190 of the naturally contaminated samples. The conventional method recovered Salmonella spp. from 622 spiked samples and 204 unspiked samples. The isolation rates from Salmonella-positive samples for the two methods were not significantly different (94.6% overall for the HGMF method and 96.7% for the conventional approach), but the HGMF results were available in only 2 to 3 days after sample receipt compared with 3 to 4 days by the conventional method. Images PMID:7059168

Rapid detection methods for viable Mycobacterium avium subspecies paratuberculosis in milk and cheese.

PubMed

Botsaris, George; Slana, Iva; Liapi, Maria; Dodd, Christine; Economides, Constantinos; Rees, Catherine; Pavlik, Ivo

2010-07-31

Mycobacterium avium subsp. paratuberculosis (MAP) may have a role in the development of Crohn's disease in humans via the consumption of contaminated milk and milk products. Detection of MAP from milk and dairy products has been reported from countries on the European continent, Argentina, the UK and Australia. In this study three different methods (quantitative real time PCR, combined phage IS900 PCR and conventional cultivation) were used to detect the presence of MAP in bulk tank milk (BTM) and cheese originating from sheep, goat and mixed milks from farms and products in Cyprus. During the first survey the presence of MAP was detected in 63 (28.6%) of cows' BTM samples by quantitative real time PCR. A second survey of BTM used a new combined phage IS900 PCR assay, and in this case MAP was detected in 50 (22.2%) samples showing a good level of agreement by both methods. None of the herds tested were known to be affected by Johne's disease and the presence of viable MAP was confirmed by conventional culture in only two cases of cows BTM. This suggests that either rapid method used is more sensitive than the conventional culture when testing raw milk samples for MAP. The two isolates recovered from BTM were identified by IS1311 PCR REA as cattle and sheep strains, respectively. In contrast when cheese samples were tested, MAP DNA was detected by quantitative real time PCR in seven (25.0%) samples (n=28). However no viable MAP was detected when either the combined phage IS900 PCR or conventional culture methods were used. Copyright 2010 Elsevier B.V. All rights reserved.

New reporting procedures based on long-term method detection levels and some considerations for interpretations of water-quality data provided by the U.S. Geological Survey National Water Quality Laboratory

USGS Publications Warehouse

Childress, Carolyn J. Oblinger; Foreman, William T.; Connor, Brooke F.; Maloney, Thomas J.

1999-01-01

This report describes the U.S. Geological Survey National Water Quality Laboratory?s approach for determining long-term method detection levels and establishing reporting levels, details relevant new reporting conventions, and provides preliminary guidance on interpreting data reported with the new conventions. At the long-term method detection level concentration, the risk of a false positive detection (analyte reported present at the long-term method detection level when not in sample) is no more than 1 percent. However, at the long-term method detection level, the risk of a false negative occurrence (analyte reported not present when present at the long-term method detection level concentration) is up to 50 percent. Because this false negative rate is too high for use as a default 'less than' reporting level, a more reliable laboratory reporting level is set at twice the determined long-term method detection level. For all methods, concentrations measured between the laboratory reporting level and the long-term method detection level will be reported as estimated concentrations. Non-detections will be censored to the laboratory reporting level. Adoption of the new reporting conventions requires a full understanding of how low-concentration data can be used and interpreted and places responsibility for using and presenting final data with the user rather than with the laboratory. Users must consider that (1) new laboratory reporting levels may differ from previously established minimum reporting levels, (2) long-term method detection levels and laboratory reporting levels may change over time, and (3) estimated concentrations are less certain than concentrations reported above the laboratory reporting level. The availability of uncensored but qualified low-concentration data for interpretation and statistical analysis is a substantial benefit to the user. A decision to censor data after they are reported from the laboratory may still be made by the user, if merited, on the basis of the intended use of the data.

Development and comparison of a real-time PCR assay for detection of Dichelobacter nodosus with culturing and conventional PCR: harmonisation between three laboratories

PubMed Central

2012-01-01

Background Ovine footrot is a contagious disease with worldwide occurrence in sheep. The main causative agent is the fastidious bacterium Dichelobacter nodosus. In Scandinavia, footrot was first diagnosed in Sweden in 2004 and later also in Norway and Denmark. Clinical examination of sheep feet is fundamental to diagnosis of footrot, but D. nodosus should also be detected to confirm the diagnosis. PCR-based detection using conventional PCR has been used at our institutes, but the method was laborious and there was a need for a faster, easier-to-interpret method. The aim of this study was to develop a TaqMan-based real-time PCR assay for detection of D. nodosus and to compare its performance with culturing and conventional PCR. Methods A D. nodosus-specific TaqMan based real-time PCR assay targeting the 16S rRNA gene was designed. The inclusivity and exclusivity (specificity) of the assay was tested using 55 bacterial and two fungal strains. To evaluate the sensitivity and harmonisation of results between different laboratories, aliquots of a single DNA preparation were analysed at three Scandinavian laboratories. The developed real-time PCR assay was compared to culturing by analysing 126 samples, and to a conventional PCR method by analysing 224 samples. A selection of PCR-products was cloned and sequenced in order to verify that they had been identified correctly. Results The developed assay had a detection limit of 3.9 fg of D. nodosus genomic DNA. This result was obtained at all three laboratories and corresponds to approximately three copies of the D. nodosus genome per reaction. The assay showed 100% inclusivity and 100% exclusivity for the strains tested. The real-time PCR assay found 54.8% more positive samples than by culturing and 8% more than conventional PCR. Conclusions The developed real-time PCR assay has good specificity and sensitivity for detection of D. nodosus, and the results are easy to interpret. The method is less time-consuming than either culturing or conventional PCR. PMID:22293440

Weak wide-band signal detection method based on small-scale periodic state of Duffing oscillator

NASA Astrophysics Data System (ADS)

Hou, Jian; Yan, Xiao-peng; Li, Ping; Hao, Xin-hong

2018-03-01

The conventional Duffing oscillator weak signal detection method, which is based on a strong reference signal, has inherent deficiencies. To address these issues, the characteristics of the Duffing oscillatorʼs phase trajectory in a small-scale periodic state are analyzed by introducing the theory of stopping oscillation system. Based on this approach, a novel Duffing oscillator weak wide-band signal detection method is proposed. In this novel method, the reference signal is discarded, and the to-be-detected signal is directly used as a driving force. By calculating the cosine function of a phase space angle, a single Duffing oscillator can be used for weak wide-band signal detection instead of an array of uncoupled Duffing oscillators. Simulation results indicate that, compared with the conventional Duffing oscillator detection method, this approach performs better in frequency detection intervals, and reduces the signal-to-noise ratio detection threshold, while improving the real-time performance of the system. Project supported by the National Natural Science Foundation of China (Grant No. 61673066).

Assessment of exposure to the Penicillium glabrum complex in cork industry using complementing methods.

PubMed

Viegas, Carla; Sabino, Raquel; Botelho, Daniel; dos Santos, Mateus; Gomes, Anita Quintal

2015-09-01

Cork oak is the second most dominant forest species in Portugal and makes this country the world leader in cork export. Occupational exposure to Chrysonilia sitophila and the Penicillium glabrum complex in cork industry is common, and the latter fungus is associated with suberosis. However, as conventional methods seem to underestimate its presence in occupational environments, the aim of our study was to see whether information obtained by polymerase chain reaction (PCR), a molecular-based method, can complement conventional findings and give a better insight into occupational exposure of cork industry workers. We assessed fungal contamination with the P. glabrum complex in three cork manufacturing plants in the outskirts of Lisbon using both conventional and molecular methods. Conventional culturing failed to detect the fungus at six sampling sites in which PCR did detect it. This confirms our assumption that the use of complementing methods can provide information for a more accurate assessment of occupational exposure to the P. glabrum complex in cork industry.

Method for detecting an image of an object

DOEpatents

Chapman, Leroy Dean; Thomlinson, William C.; Zhong, Zhong

1999-11-16

A method for detecting an absorption, refraction and scatter image of an object by independently analyzing, detecting, digitizing, and combining images acquired on a high and a low angle side of a rocking curve of a crystal analyzer. An x-ray beam which is generated by any suitable conventional apparatus can be irradiated upon either a Bragg type crystal analyzer or a Laue type crystal analyzer. Images of the absorption, refraction and scattering effects are detected, such as on an image plate, and then digitized. The digitized images are simultaneously solved, preferably on a pixel-by-pixel basis, to derive a combined visual image which has dramatically improved contrast and spatial resolution over an image acquired through conventional radiology methods.

Significance of parametric spectral ratio methods in detection and recognition of whispered speech

NASA Astrophysics Data System (ADS)

Mathur, Arpit; Reddy, Shankar M.; Hegde, Rajesh M.

2012-12-01

In this article the significance of a new parametric spectral ratio method that can be used to detect whispered speech segments within normally phonated speech is described. Adaptation methods based on the maximum likelihood linear regression (MLLR) are then used to realize a mismatched train-test style speech recognition system. This proposed parametric spectral ratio method computes a ratio spectrum of the linear prediction (LP) and the minimum variance distortion-less response (MVDR) methods. The smoothed ratio spectrum is then used to detect whispered segments of speech within neutral speech segments effectively. The proposed LP-MVDR ratio method exhibits robustness at different SNRs as indicated by the whisper diarization experiments conducted on the CHAINS and the cell phone whispered speech corpus. The proposed method also performs reasonably better than the conventional methods for whisper detection. In order to integrate the proposed whisper detection method into a conventional speech recognition engine with minimal changes, adaptation methods based on the MLLR are used herein. The hidden Markov models corresponding to neutral mode speech are adapted to the whispered mode speech data in the whispered regions as detected by the proposed ratio method. The performance of this method is first evaluated on whispered speech data from the CHAINS corpus. The second set of experiments are conducted on the cell phone corpus of whispered speech. This corpus is collected using a set up that is used commercially for handling public transactions. The proposed whisper speech recognition system exhibits reasonably better performance when compared to several conventional methods. The results shown indicate the possibility of a whispered speech recognition system for cell phone based transactions.

Supercontinuum Fourier transform spectrometry with balanced detection on a single photodiode

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

Goncharov, Vasily V.; Hall, Gregory E., E-mail: gehall@bnl.gov

We demonstrate a method of combining a supercontinuum light source with a commercial Fourier transform spectrometer, using a novel approach to dual-beam balanced detection, implemented with phase-sensitive detection on a single light detector. A 40 dB reduction in the relative intensity noise is achieved for broadband light, analogous to conventional balanced detection methods using two matched photodetectors. Unlike conventional balanced detection, however, this method exploits the time structure of the broadband source to interleave signal and reference pulse trains in the time domain, recording the broadband differential signal at the fundamental pulse repetition frequency of the supercontinuum. The method ismore » capable of real-time correction for instability in the supercontinuum spectral structure over a broad range of wavelengths and is compatible with commercially designed spectrometers. A proof-of-principle experimental setup is demonstrated for weak absorption in the 1500-1600 nm region.« less

Direct PCR - A rapid method for multiplexed detection of different serotypes of Salmonella in enriched pork meat samples.

PubMed

Chin, Wai Hoe; Sun, Yi; Høgberg, Jonas; Quyen, Than Linh; Engelsmann, Pia; Wolff, Anders; Bang, Dang Duong

2017-04-01

Salmonellosis, an infectious disease caused by Salmonella spp., is one of the most common foodborne diseases. Isolation and identification of Salmonella by conventional bacterial culture method is time consuming. In response to the demand for rapid on line or at site detection of pathogens, in this study, we developed a multiplex Direct PCR method for rapid detection of different Salmonella serotypes directly from pork meat samples without any DNA purification steps. An inhibitor-resistant Phusion Pfu DNA polymerase was used to overcome PCR inhibition. Four pairs of primers including a pair of newly designed primers targeting Salmonella spp. at subtype level were incorporated in the multiplex Direct PCR. To maximize the efficiency of the Direct PCR, the ratio between sample and dilution buffer was optimized. The sensitivity and specificity of the multiplex Direct PCR were tested using naturally contaminated pork meat samples for detecting and subtyping of Salmonella spp. Conventional bacterial culture methods were used as reference to evaluate the performance of the multiplex Direct PCR. Relative accuracy, sensitivity and specificity of 98.8%; 97.6% and 100%, respectively, were achieved by the method. Application of the multiplex Direct PCR to detect Salmonella in pork meat at slaughter reduces the time of detection from 5 to 6 days by conventional bacterial culture and serotyping methods to 14 h (including 12 h enrichment time). Furthermore, the method poses a possibility of miniaturization and integration into a point-of-need Lab-on-a-chip system for rapid online pathogen detection. Copyright © 2016 Elsevier Ltd. All rights reserved.

A New Approach to Detect Mover Position in Linear Motors Using Magnetic Sensors

PubMed Central

Paul, Sarbajit; Chang, Junghwan

2015-01-01

A new method to detect the mover position of a linear motor is proposed in this paper. This method employs a simple cheap Hall Effect sensor-based magnetic sensor unit to detect the mover position of the linear motor. With the movement of the linear motor, Hall Effect sensor modules electrically separated 120° along with the idea of three phase balanced condition (va + vb + vc = 0) are used to produce three phase signals. The amplitude of the sensor output voltage signals are adjusted to unit amplitude to minimize the amplitude errors. With the unit amplitude signals three to two phase transformation is done to reduce the three multiples of harmonic components. The final output thus obtained is converted to position data by the use of arctangent function. The measurement accuracy of the new method is analyzed by experiments and compared with the conventional two phase method. Using the same number of sensor modules as the conventional two phase method, the proposed method gives more accurate position information compared to the conventional system where sensors are separated by 90° electrical angles. PMID:26506348

Theory and practice of conventional adventitious virus testing.

PubMed

Gregersen, Jens-Peter

2011-01-01

CONFERENCE PROCEEDING Proceedings of the PDA/FDA Adventitious Viruses in Biologics: Detection and Mitigation Strategies Workshop in Bethesda, MD, USA; December 1-3, 2010 Guest Editors: Arifa Khan (Bethesda, MD), Patricia Hughes (Bethesda, MD) and Michael Wiebe (San Francisco, CA) For decades conventional tests in cell cultures and in laboratory animals have served as standard methods for broad-spectrum screening for adventitious viruses. New virus detection methods based on molecular biology have broadened and improved our knowledge about potential contaminating viruses and about the suitability of the conventional test methods. This paper summarizes and discusses practical aspects of conventional test schemes, such as detectability of various viruses, questionable or false-positive results, animal numbers needed, time and cost of testing, and applicability for rapidly changing starting materials. Strategies to improve the virus safety of biological medicinal products are proposed. The strategies should be based upon a flexible application of existing and new methods along with a scientifically based risk assessment. However, testing alone does not guarantee the absence of adventitious agents and must be accompanied by virus removing or virus inactivating process steps for critical starting materials, raw materials, and for the drug product.

Measurement of polychlorinated biphenyls in solid waste such as transformer insulation paper by supercritical fluid extraction and gas chromatography electron capture detection.

PubMed

Chikushi, Hiroaki; Fujii, Yuka; Toda, Kei

2012-09-21

In this work, a method for measuring polychlorinated biphenyls (PCBs) in contaminated solid waste was investigated. This waste includes paper that is used in electric transformers to insulate electric components. The PCBs in paper sample were extracted by supercritical fluid extraction and analyzed by gas chromatography-electron capture detection. The recoveries with this method (84-101%) were much higher than those with conventional water extraction (0.08-14%), and were comparable to those with conventional organic solvent extraction. Limit of detection was 0.0074 mg kg(-1) and measurable up to 2.5 mg kg(-1) for 0.5 g of paper sample. Data for real insulation paper by the proposed method agreed well with those by the conventional organic solvent extraction. Extraction from wood and concrete was also investigated and good performance was obtained as well as for paper samples. The supercritical fluid extraction is simpler, faster, and greener than conventional organic solvent extraction. Copyright © 2012 Elsevier B.V. All rights reserved.

A comparison of in-house real-time LAMP assays with a commercial assay for the detection of pathogenic bacteria

USDA-ARS?s Scientific Manuscript database

Molecular detection of bacterial pathogens based on LAMP methods is a faster and simpler approach than conventional culture methods. Although different LAMP-based methods for pathogenic bacterial detection are available, a systematic comparison of these different LAMP assays has not been performed. ...

Comparison on extraction yield of sennoside A and sennoside B from senna (Cassia angustifolia) using conventional and non conventional extraction techniques and their quantification using a validated HPLC-PDA detection method.

PubMed

Dhanani, Tushar; Singh, Raghuraj; Reddy, Nagaraja; Trivedi, A; Kumar, Satyanshu

2017-05-01

Senna is an important medicinal plant and is used in many Ayurvedic formulations. Dianthraquinone glucosides are the main bioactive phytochemicals present in leaves and pods of senna. The extraction efficiency in terms of yield and composition of the extract of senna prepared using both conventional (cold percolation at room temperature and refluxing) and non conventional (ultrasound and microwave assisted solvent extraction as well as supercritical fluid extraction) techniques were compared in the present study. Also a rapid reverse phase HPLC-PDA detection method was developed and validated for the simultaneous determination of sennoside A and sennoside B in the different extracts of senna leaves. Ultrasound and microwave assisted solvent extraction techniques were more effective in terms of yield and composition of the extracts compared to cold percolation at room temperature and refluxing methods of extraction.

Multi-Institutional Evaluation of Digital Tomosynthesis, Dual-Energy Radiography, and Conventional Chest Radiography for the Detection and Management of Pulmonary Nodules.

PubMed

Dobbins, James T; McAdams, H Page; Sabol, John M; Chakraborty, Dev P; Kazerooni, Ella A; Reddy, Gautham P; Vikgren, Jenny; Båth, Magnus

2017-01-01

Purpose To conduct a multi-institutional, multireader study to compare the performance of digital tomosynthesis, dual-energy (DE) imaging, and conventional chest radiography for pulmonary nodule detection and management. Materials and Methods In this binational, institutional review board-approved, HIPAA-compliant prospective study, 158 subjects (43 subjects with normal findings) were enrolled at four institutions. Informed consent was obtained prior to enrollment. Subjects underwent chest computed tomography (CT) and imaging with conventional chest radiography (posteroanterior and lateral), DE imaging, and tomosynthesis with a flat-panel imaging device. Three experienced thoracic radiologists identified true locations of nodules (n = 516, 3-20-mm diameters) with CT and recommended case management by using Fleischner Society guidelines. Five other radiologists marked nodules and indicated case management by using images from conventional chest radiography, conventional chest radiography plus DE imaging, tomosynthesis, and tomosynthesis plus DE imaging. Sensitivity, specificity, and overall accuracy were measured by using the free-response receiver operating characteristic method and the receiver operating characteristic method for nodule detection and case management, respectively. Results were further analyzed according to nodule diameter categories (3-4 mm, >4 mm to 6 mm, >6 mm to 8 mm, and >8 mm to 20 mm). Results Maximum lesion localization fraction was higher for tomosynthesis than for conventional chest radiography in all nodule size categories (3.55-fold for all nodules, P < .001; 95% confidence interval [CI]: 2.96, 4.15). Case-level sensitivity was higher with tomosynthesis than with conventional chest radiography for all nodules (1.49-fold, P < .001; 95% CI: 1.25, 1.73). Case management decisions showed better overall accuracy with tomosynthesis than with conventional chest radiography, as given by the area under the receiver operating characteristic curve (1.23-fold, P < .001; 95% CI: 1.15, 1.32). There were no differences in any specificity measures. DE imaging did not significantly affect nodule detection when paired with either conventional chest radiography or tomosynthesis. Conclusion Tomosynthesis outperformed conventional chest radiography for lung nodule detection and determination of case management; DE imaging did not show significant differences over conventional chest radiography or tomosynthesis alone. These findings indicate performance likely achievable with a range of reader expertise. © RSNA, 2016 Online supplemental material is available for this article.

Multi-Institutional Evaluation of Digital Tomosynthesis, Dual-Energy Radiography, and Conventional Chest Radiography for the Detection and Management of Pulmonary Nodules

PubMed Central

McAdams, H. Page; Sabol, John M.; Chakraborty, Dev P.; Kazerooni, Ella A.; Reddy, Gautham P.; Vikgren, Jenny; Båth, Magnus

2017-01-01

Purpose To conduct a multi-institutional, multireader study to compare the performance of digital tomosynthesis, dual-energy (DE) imaging, and conventional chest radiography for pulmonary nodule detection and management. Materials and Methods In this binational, institutional review board–approved, HIPAA-compliant prospective study, 158 subjects (43 subjects with normal findings) were enrolled at four institutions. Informed consent was obtained prior to enrollment. Subjects underwent chest computed tomography (CT) and imaging with conventional chest radiography (posteroanterior and lateral), DE imaging, and tomosynthesis with a flat-panel imaging device. Three experienced thoracic radiologists identified true locations of nodules (n = 516, 3–20-mm diameters) with CT and recommended case management by using Fleischner Society guidelines. Five other radiologists marked nodules and indicated case management by using images from conventional chest radiography, conventional chest radiography plus DE imaging, tomosynthesis, and tomosynthesis plus DE imaging. Sensitivity, specificity, and overall accuracy were measured by using the free-response receiver operating characteristic method and the receiver operating characteristic method for nodule detection and case management, respectively. Results were further analyzed according to nodule diameter categories (3–4 mm, >4 mm to 6 mm, >6 mm to 8 mm, and >8 mm to 20 mm). Results Maximum lesion localization fraction was higher for tomosynthesis than for conventional chest radiography in all nodule size categories (3.55-fold for all nodules, P < .001; 95% confidence interval [CI]: 2.96, 4.15). Case-level sensitivity was higher with tomosynthesis than with conventional chest radiography for all nodules (1.49-fold, P < .001; 95% CI: 1.25, 1.73). Case management decisions showed better overall accuracy with tomosynthesis than with conventional chest radiography, as given by the area under the receiver operating characteristic curve (1.23-fold, P < .001; 95% CI: 1.15, 1.32). There were no differences in any specificity measures. DE imaging did not significantly affect nodule detection when paired with either conventional chest radiography or tomosynthesis. Conclusion Tomosynthesis outperformed conventional chest radiography for lung nodule detection and determination of case management; DE imaging did not show significant differences over conventional chest radiography or tomosynthesis alone. These findings indicate performance likely achievable with a range of reader expertise. © RSNA, 2016 Online supplemental material is available for this article. PMID:27439324

Detection and enumeration of Salmonella enteritidis in homemade ice cream associated with an outbreak: comparison of conventional and real-time PCR methods.

PubMed

Seo, K H; Valentin-Bon, I E; Brackett, R E

2006-03-01

Salmonellosis caused by Salmonella Enteritidis (SE) is a significant cause of foodborne illnesses in the United States. Consumption of undercooked eggs and egg-containing products has been the primary risk factor for the disease. The importance of the bacterial enumeration technique has been enormously stressed because of the quantitative risk analysis of SE in shell eggs. Traditional enumeration methods mainly depend on slow and tedious most-probable-number (MPN) methods. Therefore, specific, sensitive, and rapid methods for SE quantitation are needed to collect sufficient data for risk assessment and food safety policy development. We previously developed a real-time quantitative PCR assay for the direct detection and enumeration of SE and, in this study, applied it to naturally contaminated ice cream samples with and without enrichment. The detection limit of the real-time PCR assay was determined with artificially inoculated ice cream. When applied to the direct detection and quantification of SE in ice cream, the real-time PCR assay was as sensitive as the conventional plate count method in frequency of detection. However, populations of SE derived from real-time quantitative PCR were approximately 1 log higher than provided by MPN and CFU values obtained by conventional culture methods. The detection and enumeration of SE in naturally contaminated ice cream can be completed in 3 h by this real-time PCR method, whereas the cultural enrichment method requires 5 to 7 days. A commercial immunoassay for the specific detection of SE was also included in the study. The real-time PCR assay proved to be a valuable tool that may be useful to the food industry in monitoring its processes to improve product quality and safety.

New optical method for enhanced detection of colon cancer by capsule endoscopy

NASA Astrophysics Data System (ADS)

AnkriEqually Contributed, Rinat; Peretz, Dolev; Motiei, Menachem; Sella-Tavor, Osnat; Popovtzer, Rachela

2013-09-01

PillCam®COLON capsule endoscopy (CE), a non-invasive diagnostic tool of the digestive tract, has dramatically changed the diagnostic approach and has become an attractive alternative to the conventional colonoscopy for early detection of colorectal cancer. However, despite the significant progress and non-invasive detection capability, studies have shown that its sensitivity and specificity is lower than that of conventional colonoscopy. This work presents a new optical detection method, specifically tailored to colon cancer detection and based on the well-known optical properties of immune-conjugated gold nanorods (GNRs). We show, on a colon cancer model implanted in a chick chorioallantoic membrane (CAM), that this detection method enables conclusive differentiation between cancerous and normal tissues, where neither the distance between the light source and the intestinal wall, nor the background signal, affects the monitored signal. This optical method, which can easily be integrated in CE, is expected to reduce false positive and false negative results and improve identification of tumors and micro metastases.

Double-label immunofluorescence method for simultaneous detection of adenovirus and herpes simplex virus from the eye.

PubMed

Walpita, P; Darougar, S

1989-07-01

The development and application of a double-label immunofluorescence method which has the potential to screen for single or dual infections from any site, in single shell vial cultures, is described. In this study, a total of 1,141 ocular specimens were inoculated in shell vials, centrifuged at 15,000 X g for 1 h, incubated at 37 degrees C for 48 h, and fixed in methanol at room temperature for 15 min. The virus inclusions were detected by staining with a double-label indirect immunofluorescence procedure using mixtures of appropriate first antibodies, followed by fluorescein- and rhodamine-conjugated second antibodies. Each specimen was also inoculated in parallel by the conventional virus isolation method. The sensitivity and specificity of the double-label shell vial procedure were comparable to those with the conventional method, and the former test took only 48 h to complete. The test offers a rapid and simple single-vial procedure which allows for individual or simultaneous detection of multiple pathogens. It results in savings in time and cost over the conventional virus isolation method and other shell vial procedures.

A Monocular Vision Sensor-Based Obstacle Detection Algorithm for Autonomous Robots.

PubMed

Lee, Tae-Jae; Yi, Dong-Hoon; Cho, Dong-Il Dan

2016-03-01

This paper presents a monocular vision sensor-based obstacle detection algorithm for autonomous robots. Each individual image pixel at the bottom region of interest is labeled as belonging either to an obstacle or the floor. While conventional methods depend on point tracking for geometric cues for obstacle detection, the proposed algorithm uses the inverse perspective mapping (IPM) method. This method is much more advantageous when the camera is not high off the floor, which makes point tracking near the floor difficult. Markov random field-based obstacle segmentation is then performed using the IPM results and a floor appearance model. Next, the shortest distance between the robot and the obstacle is calculated. The algorithm is tested by applying it to 70 datasets, 20 of which include nonobstacle images where considerable changes in floor appearance occur. The obstacle segmentation accuracies and the distance estimation error are quantitatively analyzed. For obstacle datasets, the segmentation precision and the average distance estimation error of the proposed method are 81.4% and 1.6 cm, respectively, whereas those for a conventional method are 57.5% and 9.9 cm, respectively. For nonobstacle datasets, the proposed method gives 0.0% false positive rates, while the conventional method gives 17.6%.

Comparison of two commercially available rapid detection methods and a conventional culture method to detect naturally occurring salmonellae on broiler carcasses

USDA-ARS?s Scientific Manuscript database

Many different screening devices and sampling methods have been used to detect the presence of naturally occurring Salmonella on commercially processed broiler carcasses. The objective of this study was to compare two commercial screening systems (BAX® and Roka®) to a standard cultural procedure use...

An Improved Azimuth Angle Estimation Method with a Single Acoustic Vector Sensor Based on an Active Sonar Detection System.

PubMed

Zhao, Anbang; Ma, Lin; Ma, Xuefei; Hui, Juan

2017-02-20

In this paper, an improved azimuth angle estimation method with a single acoustic vector sensor (AVS) is proposed based on matched filtering theory. The proposed method is mainly applied in an active sonar detection system. According to the conventional passive method based on complex acoustic intensity measurement, the mathematical and physical model of this proposed method is described in detail. The computer simulation and lake experiments results indicate that this method can realize the azimuth angle estimation with high precision by using only a single AVS. Compared with the conventional method, the proposed method achieves better estimation performance. Moreover, the proposed method does not require complex operations in frequencydomain and achieves computational complexity reduction.

[The detection of stable and unstable markers of radiation action in persons with a history of chronic irradiation using the methods of routine and differentiated staining of metaphase chromosomes].

PubMed

Shemetun, O V; Pilins'ka, M A

1998-01-01

A comparative cytogenetic observation of a group from Chernobyl NPP personnel has been carried out using conventional and G-banding staining. Detection rate of stable aberration by conventional staining to G-banding was 0.20.

[Value of combined conventional and contrast enhanced sonography in the evaluation of hepatic disorders].

PubMed

Tranquart, F; Bleuzen, A; Kissel, A

2004-06-01

To assess the value of combined conventional and contrast-material enhanced sonography for the characterization of focal liver lesions. Simultaneous imaging with grey scale and contrast enhanced US was performed in 90 patients following Levovist injection (Schering, Berlin, Germany) using the "Agent Detection Imaging" method (ADI, Siemens-Acuson, Mountain View, USA). US scanning was performed at least 4 minutes after contrast injection with review of both grayscale and contrast enhanced modes. Results for detection and characterization of lesions were compared to the selected gold standard imaging modality (CT or MRI). Final diagnoses included: 20 normal examinations, 41 patients with metastases, 6 patients with hepatocellular carcinoma, 13 patients with hemangioma, 6 patients with other benign lesions, 4 patients with cysts and 6 patients with two types of lesions. Delayed phase contrast enhanced US allowed diagnosis of all lesions except for one metastasis and all hepatocellular carcinomas. While the diagnosis of hepatoma could not be confirmed, the features suggested a malignant etiology. For 7 patients with metastases, more lesions were detected at ADI (4.9 lesions) than at conventional US (1.1 lesion). For 3 patients, CT showed more lesions than ADI US (3.3 versus 1.6 lesions). The accuracy of ADI US for differentiating between benign and malignant lesions was 98.7% compared to 49.6% for conventional US (p<0.001). The total number of lesions detected at ADI US was higher (p<0.01) than at conventional US and not significantly different from that obtained by the gold standard reference methods. Complete characterization was achieved in 92.2% of cases with ADI US compared to 59.2% with conventional US (p<0.001). Contrast-material enhanced US combined with conventional US markedly improves the diagnostic accuracy of US in terms of lesion detection and characterization.

Rapid Detection of Escherichia coli O157:H7 in Fresh Lettuce Based on Localized Surface Plasmon Resonance Combined with Immunomagnetic Separation.

PubMed

Lee, Nari; Choi, Sung-Wook; Chang, Hyun-Joo; Chun, Hyang Sook

2018-05-01

This study presents a method for rapid detection of Escherichia coli O157:H7 in fresh lettuce based on the properties of target separation and localized surface plasmon resonance of immunomagnetic nanoparticles. The multifunctional immunomagnetic nanoparticles enabling simultaneous separation and detection were prepared by synthesizing magnetic nanoparticles (ca. 10 nm in diameter) composed of an iron oxide (Fe 3 O 4 ) core and gold shell and then conjugating these nanoparticles with the anti- E. coli O157:H7 antibodies. The application of multifunctional immunomagnetic nanoparticles for detecting E. coli O157:H7 in a lettuce matrix allowed detection of the presence of <1 log CFU mL -1 without prior enrichment. In contrast, the detection limit of the conventional plating method was 2.74 log CFU mL -1 . The method, which requires no preenrichment, provides an alternative to conventional microbiological detection methods and can be used as a rapid screening tool for a large number of food samples.

Real-time explosive particle detection using a cyclone particle concentrator.

PubMed

Hashimoto, Yuichiro; Nagano, Hisashi; Takada, Yasuaki; Kashima, Hideo; Sugaya, Masakazu; Terada, Koichi; Sakairi, Minoru

2014-06-30

There is a need for more rapid methods for the detection of explosive particles. We have developed a novel real-time analysis technique for explosive particles that uses a cyclone particle concentrator. This technique can analyze sample surfaces for the presence of particles from explosives such as TNT and RDX within 3 s, which is much faster than is possible by conventional methods. Particles are detached from the sample surface with air jet pulses, and then introduced into a cyclone particle concentrator with a high pumping speed of about 80 L/min. A vaporizer placed at the bottom of the cyclone particle concentrator immediately converts the particles into a vapor. The vapor is then ionized in the atmospheric pressure chemical ionization (APCI) source of a linear ion trap mass spectrometer. An online connection between the vaporizer and a mass spectrometer enables high-speed detection within a few seconds, compared with the conventional off-line heating method that takes more than 10 s to raise the temperature of a sample filter unit. Since the configuration enriched the number density of explosive particles by about 80 times compared with that without the concentrator, a sub-ng amount of TNT particles on a surface was detectable. The detection limit of our technique is comparable with that of an explosives trace detector using ion mobility spectrometry. The technique will be beneficial for trace detection in security applications, because it detects explosive particles on the surface more speedily than conventional methods. Copyright © 2014 John Wiley & Sons, Ltd.

Recent Advances in Mycotoxin Determination for Food Monitoring via Microchip

PubMed Central

Man, Yan; Liang, Gang; Li, An; Pan, Ligang

2017-01-01

Mycotoxins are one of the main factors impacting food safety. Mycotoxin contamination has threatened the health of humans and animals. Conventional methods for the detection of mycotoxins are gas chromatography (GC) or liquid chromatography (LC) coupled with mass spectrometry (MS), or enzyme-linked immunosorbent assay (ELISA). However, all these methods are time-consuming, require large-scale instruments and skilled technicians, and consume large amounts of hazardous regents and solvents. Interestingly, a microchip requires less sample consumption and short analysis time, and can realize the integration, miniaturization, and high-throughput detection of the samples. Hence, the application of a microchip for the detection of mycotoxins can make up for the deficiency of the conventional detection methods. This review focuses on the application of a microchip to detect mycotoxins in foods. The toxicities of mycotoxins and the materials of the microchip are firstly summarized in turn. Then the application of a microchip that integrates various kinds of detection methods (optical, electrochemical, photo-electrochemical, and label-free detection) to detect mycotoxins is reviewed in detail. Finally, challenges and future research directions in the development of a microchip to detect mycotoxins are previewed. PMID:29036884

Electronic method for autofluorography of macromolecules on two-D matrices

DOEpatents

Davidson, Jackson B.; Case, Arthur L.

1983-01-01

A method for detecting, localizing, and quantifying macromolecules contained in a two-dimensional matrix is provided which employs a television-based position sensitive detection system. A molecule-containing matrix may be produced by conventional means to produce spots of light at the molecule locations which are detected by the television system. The matrix, such as a gel matrix, is exposed to an electronic camera system including an image-intensifier and secondary electron conduction camera capable of light integrating times of many minutes. A light image stored in the form of a charge image on the camera tube target is scanned by conventional television techniques, digitized, and stored in a digital memory. Intensity of any point on the image may be determined from the number at the memory address of the point. The entire image may be displayed on a television monitor for inspection and photographing or individual spots may be analyzed through selected readout of the memory locations. Compared to conventional film exposure methods, the exposure time may be reduced 100-1000 times.

Hydrogen peroxide test for intraoperative bile leak detection.

PubMed

Trehan, V; Rao, Pankaj P; Naidu, C S; Sharma, Anuj K; Singh, A K; Sharma, Sanjay; Gaur, Amit; Kulkarni, S V; Pathak, N

2017-07-01

Bile leakage (BL) is a common complication following liver surgery, ranging from 3 to 27% in different series. To reduce the incidence of post-operative BL various BL tests have been applied since ages, but no method is foolproof and every method has their own limitations. In this study we used a relatively simpler technique to detect the BL intra-operatively. Topical application of 1.5% diluted hydrogen peroxide (H 2 O 2 ) was used to detect the BL from cut surface of liver and we compared this with conventional saline method to know the efficacy. A total of 31 patients included all patients who underwent liver resection and donor hepatectomies as part of Living Donor Liver Transplantation. After complete liver resection, the conventional saline test followed by topical diluted 1.5% H 2 O 2 test was performed on all. A BL was demonstrated in 11 patients (35.48%) by the conventional saline method and in 19 patients (61.29%) by H 2 O 2 method. Statistically compared by Wilcoxon signed-rank test showed significant difference ( P  = 0.014) for minor liver resections group and ( P  = 0.002) for major liver resections group. The topical application of H 2 O 2 is a simple and effective method of detection of BL from cut surface of liver. It is an easy, non-invasive, cheap, less time consuming, reproducible, and sensitive technique with no obvious disadvantages.

[Sensitivity and specificity of nested PCR pyrosequencing in hepatitis B virus drug resistance gene testing].

PubMed

Sun, Shumei; Zhou, Hao; Zhou, Bin; Hu, Ziyou; Hou, Jinlin; Sun, Jian

2012-05-01

To evaluate the sensitivity and specificity of nested PCR combined with pyrosequencing in the detection of HBV drug-resistance gene. RtM204I (ATT) mutant and rtM204 (ATG) nonmutant plasmids mixed at different ratios were detected for mutations using nested-PCR combined with pyrosequencing, and the results were compared with those by conventional PCR pyrosequencing to analyze the linearity and consistency of the two methods. Clinical specimens with different viral loads were examined for drug-resistant mutations using nested PCR pyrosequencing and nested PCR combined with dideoxy sequencing (Sanger) for comparison of the detection sensitivity and specificity. The fitting curves demonstrated good linearity of both conventional PCR pyrosequencing and nested PCR pyrosequencing (R(2)>0.99, P<0.05). Nested PCR showed a better consistency with the predicted value than conventional PCR, and was superior to conventional PCR for detection of samples containing 90% mutant plasmid. In the detection of clinical specimens, Sanger sequencing had a significantly lower sensitivity than nested PCR pyrosequencing (92% vs 100%, P<0.01). The detection sensitivity of Sanger sequencing varied with the viral loads, especially in samples with low viral copies (HBV DNA ≤3log10 copies/ml), where the sensitivity was 78%, significantly lower than that of pyrosequencing (100%, P<0.01). Neither of the two methods yielded positive results for the negative control samples, suggesting their good specificity. Compared with nested PCR and Sanger sequencing method, nested PCR pyrosequencing has a higher sensitivity especially in clinical specimens with low viral copies, which can be important for early detection of HBV mutant strains and hence more effective clinical management.

[Usefulness of conventional polymerase chain reaction for the detection of Mycoplasma hominis, Ureaplasma spp. and Trichomonas vaginalis in female outpatient's genital samples].

PubMed

Alarcón, Gonzalo; Barraza, Gabriela; Vera, Andrea; Wozniak, Aniela; García, Patricia

2016-02-01

Trichomonas vaginalis, Mycoplasma hominis and Ureaplasma spp. are microorganisms responsible for genitourinary and pregnancy pathologies. Nucleic acid amplification methods have shown several advantages, but have not been widely studied for the detection of these microorganisms. To implement a conventional polymerase chain reaction (PCR) for the detection of the microorganisms and to compare its results versus the methods currently used at our laboratory. 91 available samples were processed by PCR, culture (M. hominis y Ureaplasma spp.) and wet mount (T vaginalis). Results were compared and statistically analyzed by kappa agreement test. 85, 80 and 87 samples resulted in agreement for the detection of M. hominis, Ureaplasma spp. y T. vaginalis, respectively. For M. hominis and Ureaplasma spp., agreement was substantial, whereas for T. vaginalis it was moderate, however, for the latter, PCR detected more cases than wet mount. We recommend the implementation of PCR for detection of T. vaginalis whereas culture kit is still a useful method for the other microorganisms.

An Improved Azimuth Angle Estimation Method with a Single Acoustic Vector Sensor Based on an Active Sonar Detection System

PubMed Central

Zhao, Anbang; Ma, Lin; Ma, Xuefei; Hui, Juan

2017-01-01

In this paper, an improved azimuth angle estimation method with a single acoustic vector sensor (AVS) is proposed based on matched filtering theory. The proposed method is mainly applied in an active sonar detection system. According to the conventional passive method based on complex acoustic intensity measurement, the mathematical and physical model of this proposed method is described in detail. The computer simulation and lake experiments results indicate that this method can realize the azimuth angle estimation with high precision by using only a single AVS. Compared with the conventional method, the proposed method achieves better estimation performance. Moreover, the proposed method does not require complex operations in frequency-domain and achieves computational complexity reduction. PMID:28230763

A novel method for accurate needle-tip identification in trans-rectal ultrasound-based high-dose-rate prostate brachytherapy.

PubMed

Zheng, Dandan; Todor, Dorin A

2011-01-01

In real-time trans-rectal ultrasound (TRUS)-based high-dose-rate prostate brachytherapy, the accurate identification of needle-tip position is critical for treatment planning and delivery. Currently, needle-tip identification on ultrasound images can be subject to large uncertainty and errors because of ultrasound image quality and imaging artifacts. To address this problem, we developed a method based on physical measurements with simple and practical implementation to improve the accuracy and robustness of needle-tip identification. Our method uses measurements of the residual needle length and an off-line pre-established coordinate transformation factor, to calculate the needle-tip position on the TRUS images. The transformation factor was established through a one-time systematic set of measurements of the probe and template holder positions, applicable to all patients. To compare the accuracy and robustness of the proposed method and the conventional method (ultrasound detection), based on the gold-standard X-ray fluoroscopy, extensive measurements were conducted in water and gel phantoms. In water phantom, our method showed an average tip-detection accuracy of 0.7 mm compared with 1.6 mm of the conventional method. In gel phantom (more realistic and tissue-like), our method maintained its level of accuracy while the uncertainty of the conventional method was 3.4mm on average with maximum values of over 10mm because of imaging artifacts. A novel method based on simple physical measurements was developed to accurately detect the needle-tip position for TRUS-based high-dose-rate prostate brachytherapy. The method demonstrated much improved accuracy and robustness over the conventional method. Copyright © 2011 American Brachytherapy Society. Published by Elsevier Inc. All rights reserved.

Scanning Electron Microscope-Cathodoluminescence Analysis of Rare-Earth Elements in Magnets.

PubMed

Imashuku, Susumu; Wagatsuma, Kazuaki; Kawai, Jun

2016-02-01

Scanning electron microscope-cathodoluminescence (SEM-CL) analysis was performed for neodymium-iron-boron (NdFeB) and samarium-cobalt (Sm-Co) magnets to analyze the rare-earth elements present in the magnets. We examined the advantages of SEM-CL analysis over conventional analytical methods such as SEM-energy-dispersive X-ray (EDX) spectroscopy and SEM-wavelength-dispersive X-ray (WDX) spectroscopy for elemental analysis of rare-earth elements in NdFeB magnets. Luminescence spectra of chloride compounds of elements in the magnets were measured by the SEM-CL method. Chloride compounds were obtained by the dropwise addition of hydrochloric acid on the magnets followed by drying in vacuum. Neodymium, praseodymium, terbium, and dysprosium were separately detected in the NdFeB magnets, and samarium was detected in the Sm-Co magnet by the SEM-CL method. In contrast, it was difficult to distinguish terbium and dysprosium in the NdFeB magnet with a dysprosium concentration of 1.05 wt% by conventional SEM-EDX analysis. Terbium with a concentration of 0.02 wt% in an NdFeB magnet was detected by SEM-CL analysis, but not by conventional SEM-WDX analysis. SEM-CL analysis is advantageous over conventional SEM-EDX and SEM-WDX analyses for detecting trace rare-earth elements in NdFeB magnets, particularly dysprosium and terbium.

A Monocular Vision Sensor-Based Obstacle Detection Algorithm for Autonomous Robots

PubMed Central

Lee, Tae-Jae; Yi, Dong-Hoon; Cho, Dong-Il “Dan”

2016-01-01

This paper presents a monocular vision sensor-based obstacle detection algorithm for autonomous robots. Each individual image pixel at the bottom region of interest is labeled as belonging either to an obstacle or the floor. While conventional methods depend on point tracking for geometric cues for obstacle detection, the proposed algorithm uses the inverse perspective mapping (IPM) method. This method is much more advantageous when the camera is not high off the floor, which makes point tracking near the floor difficult. Markov random field-based obstacle segmentation is then performed using the IPM results and a floor appearance model. Next, the shortest distance between the robot and the obstacle is calculated. The algorithm is tested by applying it to 70 datasets, 20 of which include nonobstacle images where considerable changes in floor appearance occur. The obstacle segmentation accuracies and the distance estimation error are quantitatively analyzed. For obstacle datasets, the segmentation precision and the average distance estimation error of the proposed method are 81.4% and 1.6 cm, respectively, whereas those for a conventional method are 57.5% and 9.9 cm, respectively. For nonobstacle datasets, the proposed method gives 0.0% false positive rates, while the conventional method gives 17.6%. PMID:26938540

A broad range assay for rapid detection and etiologic characterization of bacterial meningitis: performance testing in samples from sub-Sahara.

PubMed

Won, Helen; Yang, Samuel; Gaydos, Charlotte; Hardick, Justin; Ramachandran, Padmini; Hsieh, Yu-Hsiang; Kecojevic, Alexander; Njanpop-Lafourcade, Berthe-Marie; Mueller, Judith E; Tameklo, Tsidi Agbeko; Badziklou, Kossi; Gessner, Bradford D; Rothman, Richard E

2012-09-01

This study aimed to conduct a pilot evaluation of broad-based multiprobe polymerase chain reaction (PCR) in clinical cerebrospinal fluid (CSF) samples compared to local conventional PCR/culture methods used for bacterial meningitis surveillance. A previously described PCR consisting of initial broad-based detection of Eubacteriales by a universal probe, followed by Gram typing, and pathogen-specific probes was designed targeting variable regions of the 16S rRNA gene. The diagnostic performance of the 16S rRNA assay in "127 CSF samples was evaluated in samples from patients from Togo, Africa, by comparison to conventional PCR/culture methods. Our probes detected Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae. Uniprobe sensitivity and specificity versus conventional PCR were 100% and 54.6%, respectively. Sensitivity and specificity of uniprobe versus culture methods were 96.5% and 52.5%, respectively. Gram-typing probes correctly typed 98.8% (82/83) and pathogen-specific probes identified 96.4% (80/83) of the positives. This broad-based PCR algorithm successfully detected and provided species level information for multiple bacterial meningitis agents in clinical samples. Copyright © 2012 Elsevier Inc. All rights reserved.

A broad range assay for rapid detection and etiologic characterization of bacterial meningitis: performance testing in samples from sub-Sahara☆, ☆☆,★

PubMed Central

Won, Helen; Yang, Samuel; Gaydos, Charlotte; Hardick, Justin; Ramachandran, Padmini; Hsieh, Yu-Hsiang; Kecojevic, Alexander; Njanpop-Lafourcade, Berthe-Marie; Mueller, Judith E.; Tameklo, Tsidi Agbeko; Badziklou, Kossi; Gessner, Bradford D.; Rothman, Richard E.

2012-01-01

This study aimed to conduct a pilot evaluation of broad-based multiprobe polymerase chain reaction (PCR) in clinical cerebrospinal fluid (CSF) samples compared to local conventional PCR/culture methods used for bacterial meningitis surveillance. A previously described PCR consisting of initial broad-based detection of Eubacteriales by a universal probe, followed by Gram typing, and pathogen-specific probes was designed targeting variable regions of the 16S rRNA gene. The diagnostic performance of the 16S rRNA assay in “”127 CSF samples was evaluated in samples from patients from Togo, Africa, by comparison to conventional PCR/culture methods. Our probes detected Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae. Uniprobe sensitivity and specificity versus conventional PCR were 100% and 54.6%, respectively. Sensitivity and specificity of uniprobe versus culture methods were 96.5% and 52.5%, respectively. Gram-typing probes correctly typed 98.8% (82/83) and pathogen-specific probes identified 96.4% (80/83) of the positives. This broad-based PCR algorithm successfully detected and provided species level information for multiple bacterial meningitis agents in clinical samples. PMID:22809694

Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing

PubMed Central

Espy, M. J.; Uhl, J. R.; Sloan, L. M.; Buckwalter, S. P.; Jones, M. F.; Vetter, E. A.; Yao, J. D. C.; Wengenack, N. L.; Rosenblatt, J. E.; Cockerill, F. R.; Smith, T. F.

2006-01-01

Real-time PCR has revolutionized the way clinical microbiology laboratories diagnose many human microbial infections. This testing method combines PCR chemistry with fluorescent probe detection of amplified product in the same reaction vessel. In general, both PCR and amplified product detection are completed in an hour or less, which is considerably faster than conventional PCR detection methods. Real-time PCR assays provide sensitivity and specificity equivalent to that of conventional PCR combined with Southern blot analysis, and since amplification and detection steps are performed in the same closed vessel, the risk of releasing amplified nucleic acids into the environment is negligible. The combination of excellent sensitivity and specificity, low contamination risk, and speed has made real-time PCR technology an appealing alternative to culture- or immunoassay-based testing methods for diagnosing many infectious diseases. This review focuses on the application of real-time PCR in the clinical microbiology laboratory. PMID:16418529

Damage diagnosis algorithm using a sequential change point detection method with an unknown distribution for damage

NASA Astrophysics Data System (ADS)

Noh, Hae Young; Rajagopal, Ram; Kiremidjian, Anne S.

2012-04-01

This paper introduces a damage diagnosis algorithm for civil structures that uses a sequential change point detection method for the cases where the post-damage feature distribution is unknown a priori. This algorithm extracts features from structural vibration data using time-series analysis and then declares damage using the change point detection method. The change point detection method asymptotically minimizes detection delay for a given false alarm rate. The conventional method uses the known pre- and post-damage feature distributions to perform a sequential hypothesis test. In practice, however, the post-damage distribution is unlikely to be known a priori. Therefore, our algorithm estimates and updates this distribution as data are collected using the maximum likelihood and the Bayesian methods. We also applied an approximate method to reduce the computation load and memory requirement associated with the estimation. The algorithm is validated using multiple sets of simulated data and a set of experimental data collected from a four-story steel special moment-resisting frame. Our algorithm was able to estimate the post-damage distribution consistently and resulted in detection delays only a few seconds longer than the delays from the conventional method that assumes we know the post-damage feature distribution. We confirmed that the Bayesian method is particularly efficient in declaring damage with minimal memory requirement, but the maximum likelihood method provides an insightful heuristic approach.

Rapid detection of microbes in the dialysis solution by the microcolony fluorescence staining method (Millflex quantum).

PubMed

Osono, Eiichi; Kobayashi, Eiko; Inoue, Yuki; Honda, Kazumi; Kumagai, Takuya; Negishi, Hideki; Okamatsu, Kentaro; Ichimura, Kyoko; Kamano, Chisako; Suzuki, Fumi; Norose, Yoshihiko; Takahashi, Megumi; Takaku, Shun; Fujioka, Noriaki; Hayama, Naoaki; Takizawa, Hideaki

2014-01-01

A chemiluminescence system, Milliflex Quantum (MFQ), to detect microcolonies, has been used in the pharmaceutical field. In this study, we investigated aquatic bacteria in hemodialysis solutions sampled from bioburden areas in 4 dialysis faculties. Using MFQ, microcolonies could be detected after a short incubation period. The colony count detected with MFQ after a 48-hour incubation was 92% ± 39%, compared to that after the conventionally used 7-14-day incubation period; in addition, the results also showed a linear correlation. Moreover, MFQ-based analysis allowed the visualization of damaged cells and of the high density due to the excessive amount of bacteria. These results suggested that MFQ had adequate sensitivity to detect microbacteria in dialysis solutions, and it was useful for validating the conditions of conventional culture methods.

Comparison of Conventional PCR, Multiplex PCR, and Loop-Mediated Isothermal Amplification Assays for Rapid Detection of Arcobacter Species

PubMed Central

Wang, Xiaoyu; Seo, Dong Joo; Lee, Min Hwa

2014-01-01

This study aimed to develop a loop-mediated isothermal amplification (LAMP) method for the rapid detection of Arcobacter species. Specific primers targeting the 23S ribosomal RNA gene were used to detect Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. The specificity of the LAMP primer set was assessed using DNA samples from a panel of Arcobacter and Campylobacter species, and the sensitivity was determined using serial dilutions of Arcobacter species cultures. LAMP showed a 10- to 1,000-fold-higher sensitivity than multiplex PCR, with a detection limit of 2 to 20 CFU per reaction in vitro. Whereas multiplex PCR showed cross-reactivity with Campylobacter species, the LAMP method developed in this study was more sensitive and reliable than conventional PCR or multiplex PCR for the detection of Arcobacter species. PMID:24478488

Near Infrared Imaging as a Diagnostic Tool for Detecting Enamel Demineralization: An in vivo Study

NASA Astrophysics Data System (ADS)

Lucas, Seth Adam

Background and Objectives: For decades there has been an effort to develop alternative optical methods of imaging dental decay utilizing non-ionizing radiation methods. The purpose of this in-vivo study was to demonstrate whether NIR can be used as a diagnostic tool to evaluate dental caries and to compare the sensitivity and specificity of this method with that of conventional methods, including bitewing x-rays and visual inspection. Materials and Methods: 31 test subjects (n=31) from the UCSF orthodontic clinic undergoing orthodontic treatment with planned premolar extractions were recruited. Calibrated examiners performed caries detection examinations using conventional methods: bitewing radiographs and visual inspection. These findings were compared with the results from NIR examinations: transillumination and reflectance. To confirm the results found in the two different detection methods, a gold standard was used. After teeth were extracted, polarized light microscopy and transverse microradiography were performed. Results: A total of 87 premolars were used in the study. NIR identified the occlusal lesions with a sensitivity of 71% and a specificity of 77%, whereas, the visual examination had a sensitivity of only 40% and a specifity of 39%. For interproximal lesions halfway to DEJ, specificity remained constant, but sensitivity improved to 100% for NIR and 75% for x-rays. Conclusions: The results of this preliminary study demonstrate that NIR is just as effective at detecting enamel interproximal lesions as standard dental x-rays. NIR was more effective at detecting occlusal lesions than visual examination alone. NIR shows promise as an alternative diagnostic tool to the conventional methods of x-rays and visual examination and provides a non-ionizing radiation technique.

Early detection of materials degradation

NASA Astrophysics Data System (ADS)

Meyendorf, Norbert

2017-02-01

Lightweight components for transportation and aerospace applications are designed for an estimated lifecycle, taking expected mechanical and environmental loads into account. The main reason for catastrophic failure of components within the expected lifecycle are material inhomogeneities, like pores and inclusions as origin for fatigue cracks, that have not been detected by NDE. However, material degradation by designed or unexpected loading conditions or environmental impacts can accelerate the crack initiation or growth. Conventional NDE methods are usually able to detect cracks that are formed at the end of the degradation process, but methods for early detection of fatigue, creep, and corrosion are still a matter of research. For conventional materials ultrasonic, electromagnetic, or thermographic methods have been demonstrated as promising. Other approaches are focused to surface damage by using optical methods or characterization of the residual surface stresses that can significantly affect the creation of fatigue cracks. For conventional metallic materials, material models for nucleation and propagation of damage have been successfully applied for several years. Material microstructure/property relations are well established and the effect of loading conditions on the component life can be simulated. For advanced materials, for example carbon matrix composites or ceramic matrix composites, the processes of nucleation and propagation of damage is still not fully understood. For these materials NDE methods can not only be used for the periodic inspections, but can significantly contribute to the material scientific knowledge to understand and model the behavior of composite materials.

Teschoviruses as Indicators of Porcine Fecal Contamination of Surface Water

PubMed Central

Jiménez-Clavero, Miguel Angel; Fernández, Carlos; Ortiz, José Antonio; Pro, Javier; Carbonell, Gregoria; Tarazona, José Vicente; Roblas, Neftalí; Ley, Victoria

2003-01-01

Teschoviruses specifically infect pigs and are shed in pig feces. Hence, their presence in water should indicate contamination with pig fecal residues. To assess this hypothesis, we have developed a real-time reverse transcriptase PCR (RT-PCR) method that allows the quantitative detection of pig teschovirus (PTV) RNA. The method is able to detect 92 fg of PTV RNA per ml of sample. Using this method, we have detected the presence of PTV RNA in water and fecal samples from all pig farms examined (n = 5). Feces from other animal species (cattle, sheep, and goats) were negative in this test. To compare the PTV RNA detection method with conventional chemical determinations currently in use for evaluation of water contamination, we analyzed water samples collected downstream from a pig slurry spillage site. We have found a positive correlation within both types of determinations. The sensitivity of the PTV detection assay was similar to that achieved by unspecific organic matter determination and superior to all other conventional chemical analyses performed. Furthermore, the new method is highly specific, revealing the porcine origin of the contamination, a feature that is lacking in currently available methods for the assessment of water contamination. PMID:14532098

Islanding detection technique using wavelet energy in grid-connected PV system

NASA Astrophysics Data System (ADS)

Kim, Il Song

2016-08-01

This paper proposes a new islanding detection method using wavelet energy in a grid-connected photovoltaic system. The method detects spectral changes in the higher-frequency components of the point of common coupling voltage and obtains wavelet coefficients by multilevel wavelet analysis. The autocorrelation of the wavelet coefficients can clearly identify islanding detection, even in the variations of the grid voltage harmonics during normal operating conditions. The advantage of the proposed method is that it can detect islanding condition the conventional under voltage/over voltage/under frequency/over frequency methods fail to detect. The theoretical method to obtain wavelet energies is evolved and verified by the experimental result.

Ultratrace level determination and quantitative analysis of kidney injury biomarkers in patient samples attained by zinc oxide nanorods

NASA Astrophysics Data System (ADS)

Singh, Manpreet; Alabanza, Anginelle; Gonzalez, Lorelis E.; Wang, Weiwei; Reeves, W. Brian; Hahm, Jong-In

2016-02-01

Determining ultratrace amounts of protein biomarkers in patient samples in a straightforward and quantitative manner is extremely important for early disease diagnosis and treatment. Here, we successfully demonstrate the novel use of zinc oxide nanorods (ZnO NRs) in the ultrasensitive and quantitative detection of two acute kidney injury (AKI)-related protein biomarkers, tumor necrosis factor (TNF)-α and interleukin (IL)-8, directly from patient samples. We first validate the ZnO NRs-based IL-8 results via comparison with those obtained from using a conventional enzyme-linked immunosorbent method in samples from 38 individuals. We further assess the full detection capability of the ZnO NRs-based technique by quantifying TNF-α, whose levels in human urine are often below the detection limits of conventional methods. Using the ZnO NR platforms, we determine the TNF-α concentrations of all 46 patient samples tested, down to the fg per mL level. Subsequently, we screen for TNF-α levels in approximately 50 additional samples collected from different patient groups in order to demonstrate a potential use of the ZnO NRs-based assay in assessing cytokine levels useful for further clinical monitoring. Our research efforts demonstrate that ZnO NRs can be straightforwardly employed in the rapid, ultrasensitive, quantitative, and simultaneous detection of multiple AKI-related biomarkers directly in patient urine samples, providing an unparalleled detection capability beyond those of conventional analysis methods. Additional key advantages of the ZnO NRs-based approach include a fast detection speed, low-volume assay condition, multiplexing ability, and easy automation/integration capability to existing fluorescence instrumentation. Therefore, we anticipate that our ZnO NRs-based detection method will be highly beneficial for overcoming the frequent challenges in early biomarker development and treatment assessment, pertaining to the facile and ultrasensitive quantification of hard-to-trace biomolecules.Determining ultratrace amounts of protein biomarkers in patient samples in a straightforward and quantitative manner is extremely important for early disease diagnosis and treatment. Here, we successfully demonstrate the novel use of zinc oxide nanorods (ZnO NRs) in the ultrasensitive and quantitative detection of two acute kidney injury (AKI)-related protein biomarkers, tumor necrosis factor (TNF)-α and interleukin (IL)-8, directly from patient samples. We first validate the ZnO NRs-based IL-8 results via comparison with those obtained from using a conventional enzyme-linked immunosorbent method in samples from 38 individuals. We further assess the full detection capability of the ZnO NRs-based technique by quantifying TNF-α, whose levels in human urine are often below the detection limits of conventional methods. Using the ZnO NR platforms, we determine the TNF-α concentrations of all 46 patient samples tested, down to the fg per mL level. Subsequently, we screen for TNF-α levels in approximately 50 additional samples collected from different patient groups in order to demonstrate a potential use of the ZnO NRs-based assay in assessing cytokine levels useful for further clinical monitoring. Our research efforts demonstrate that ZnO NRs can be straightforwardly employed in the rapid, ultrasensitive, quantitative, and simultaneous detection of multiple AKI-related biomarkers directly in patient urine samples, providing an unparalleled detection capability beyond those of conventional analysis methods. Additional key advantages of the ZnO NRs-based approach include a fast detection speed, low-volume assay condition, multiplexing ability, and easy automation/integration capability to existing fluorescence instrumentation. Therefore, we anticipate that our ZnO NRs-based detection method will be highly beneficial for overcoming the frequent challenges in early biomarker development and treatment assessment, pertaining to the facile and ultrasensitive quantification of hard-to-trace biomolecules. Electronic supplementary information (ESI) available: Typical SEM images of the ZnO NRs used in the biomarker assays are provided in Fig. S1. See DOI: 10.1039/c5nr08706f

Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran.

PubMed

Abdoli, Amir; Dalimi, Abdolhossein; Soltanghoraee, Haleh; Ghaffarifar, Fatemeh

2016-12-01

Toxoplasma gondii is one of the most common zoonotic parasitic diseases in human and warm-blooded animals worldwide. Birds are one of important intermediate hosts of T. gondii . The aim of this study is molecular detection of T. gondii in the house sparrow by LAMP and PCR methods in Tehran, Iran. A total 200 sparrows were captured in different regions of Tehran. DNA was extracted from tissue samples of each sparrow. LAMP and conventional PCR assays were carried out with a set of primers to detect the 529 bp fragment of T. gondii . LAMP and PCR were detected T. gondii from 17 (8.5 %) and 15 (7.5 %) of 200 sparrows respectively. These results indicated that sensitivity of LAMP was higher than conventional PCR. In our knowledge, this study is the first report of detection of T. gondii by LAMP method in bird hosts. Also, these findings provided an insight into epidemiological pattern of T. gondii infection in sparrow in Iran.

On-line high-speed rail defect detection : part II.

DOT National Transportation Integrated Search

2012-03-01

The objectives of this project were (1) to improve the defect detection reliability and (2) to improve the inspection speed of conventional rail defect detection methods. The prototype developed in this work uses noncontact transducers, ultrasonic gu...

Novel and highly sensitive sybr® green real-time pcr for poxvirus detection in odontocete cetaceans.

PubMed

Sacristán, Carlos; Luiz Catão-Dias, José; Ewbank, Ana Carolina; Machado, Eduardo Ferreira; Neves, Elena; Santos-Neto, Elitieri Batista; Azevedo, Alexandre; Laison-Brito, José; De Castilho, Pedro Volkmer; Daura-Jorge, Fábio Gonçalves; Simões-Lopes, Paulo César; Carballo, Matilde; García-Párraga, Daniel; Manuel Sánchez-Vizcaíno, José; Esperón, Fernando

2018-06-08

Poxviruses are emerging pathogens in cetaceans, temporarily named 'Cetaceanpoxvirus' (CePV, family Poxviridae), classified into two main lineages: CePV-1 in odontocetes and CePV-2 in mysticetes. Only a few studies performed the molecular detection of CePVs, based on DNA-polymerase gene and/or DNA-topoisomerase I gene amplification. Herein we describe a new real-time PCR assay based on SYBR ® Green and a new primer set to detect a 150 bp fragment of CePV DNA-polymerase gene, also effective for conventional PCR detection. The novel real-time PCR was able to detect 5 up to 5 × 10 6 copies per reaction of a cloned positive control. Both novel PCR methods were 1000 to 100,000-fold more sensitive than those previously described in the literature. Samples of characteristic poxvirus skin lesions ('tattoo') from one Risso's dolphin (Grampus griseus), two striped dolphins (Stenella coeruleoalba) and two Guiana dolphins (Sotalia guianensis) were all positive to both our novel real time- and conventional PCR methods, even though three of these animals (a Risso's dolphin, a striped dolphin, and a Guiana dolphin) were previously negative to the conventional PCRs previously available. To our knowledge, this is the first real-time PCR detection method for Cetaceanpoxvirus, a much more sensitive tool for the detection of CePV-1 infections. Copyright © 2018 Elsevier B.V. All rights reserved.

A novel method of multiple nucleic acid detection: Real-time RT-PCR coupled with probe-melting curve analysis.

PubMed

Han, Yang; Hou, Shao-Yang; Ji, Shang-Zhi; Cheng, Juan; Zhang, Meng-Yue; He, Li-Juan; Ye, Xiang-Zhong; Li, Yi-Min; Zhang, Yi-Xuan

2017-11-15

A novel method, real-time reverse transcription PCR (real-time RT-PCR) coupled with probe-melting curve analysis, has been established to detect two kinds of samples within one fluorescence channel. Besides a conventional TaqMan probe, this method employs another specially designed melting-probe with a 5' terminus modification which meets the same label with the same fluorescent group. By using an asymmetric PCR method, the melting-probe is able to detect an extra sample in the melting stage effectively while it almost has little influence on the amplification detection. Thus, this method allows the availability of united employment of both amplification stage and melting stage for detecting samples in one reaction. The further demonstration by simultaneous detection of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) in one channel as a model system is presented in this essay. The sensitivity of detection by real-time RT-PCR coupled with probe-melting analysis was proved to be equal to that detected by conventional real-time RT-PCR. Because real-time RT-PCR coupled with probe-melting analysis can double the detection throughputs within one fluorescence channel, it is expected to be a good solution for the problem of low-throughput in current real-time PCR. Copyright © 2017 Elsevier Inc. All rights reserved.

Electronic method for autofluorography of macromolecules on two-D matrices. [Patent application

DOEpatents

Davidson, J.B.; Case, A.L.

1981-12-30

A method for detecting, localizing, and quantifying macromolecules contained in a two-dimensional matrix is provided which employs a television-based position sensitive detection system. A molecule-containing matrix may be produced by conventional means to produce spots of light at the molecule locations which are detected by the television system. The matrix, such as a gel matrix, is exposed to an electronic camera system including an image-intensifier and secondary electron conduction camera capable of light integrating times of many minutes. A light image stored in the form of a charge image on the camera tube target is scanned by conventional television techniques, digitized, and stored in a digital memory. Intensity of any point on the image may be determined from the number at the memory address of the point. The entire image may be displayed on a television monitor for inspection and photographing or individual spots may be analyzed through selected readout of the memory locations. Compared to conventional film exposure methods, the exposure time may be reduced 100 to 1000 times.

Comparisons between conventional optical imaging and parametric indirect microscopic imaging on human skin detection

NASA Astrophysics Data System (ADS)

Liu, Guoyan; Gao, Kun; Liu, Xuefeng; Ni, Guoqiang

2016-10-01

We report a new method, polarization parameters indirect microscopic imaging with a high transmission infrared light source, to detect the morphology and component of human skin. A conventional reflection microscopic system is used as the basic optical system, into which a polarization-modulation mechanics is inserted and a high transmission infrared light source is utilized. The near-field structural characteristics of human skin can be delivered by infrared waves and material coupling. According to coupling and conduction physics, changes of the optical wave parameters can be calculated and curves of the intensity of the image can be obtained. By analyzing the near-field polarization parameters in nanoscale, we can finally get the inversion images of human skin. Compared with the conventional direct optical microscope, this method can break diffraction limit and achieve a super resolution of sub-100nm. Besides, the method is more sensitive to the edges, wrinkles, boundaries and impurity particles.

A rigorous assessment of tree height measurements obtained using airborne LIDAR and conventional field methods.

Treesearch

Hans-Erik Andersen; Stephen E. Reutebuch; Robert J. McGaughey

2006-01-01

Tree height is an important variable in forest inventory programs but is typically time-consuming and costly to measure in the field using conventional techniques. Airborne light detection and ranging (LIDAR) provides individual tree height measurements that are highly correlated with field-derived measurements, but the imprecision of conventional field techniques does...

Diagnosis of Cetacean morbillivirus: A sensitive one step real time RT fast-PCR method based on SYBR(®) Green.

PubMed

Sacristán, Carlos; Carballo, Matilde; Muñoz, María Jesús; Bellière, Edwige Nina; Neves, Elena; Nogal, Verónica; Esperón, Fernando

2015-12-15

Cetacean morbillivirus (CeMV) (family Paramyxoviridae, genus Morbillivirus) is considered the most pathogenic virus of cetaceans. It was first implicated in the bottlenose dolphin (Tursiops truncatus) mass stranding episode along the Northwestern Atlantic coast in the late 1980s, and in several more recent worldwide epizootics in different Odontoceti species. This study describes a new one step real-time reverse transcription fast polymerase chain reaction (real-time RT-fast PCR) method based on SYBR(®) Green to detect a fragment of the CeMV fusion protein gene. This primer set also works for conventional RT-PCR diagnosis. This method detected and identified all three well-characterized strains of CeMV: porpoise morbillivirus (PMV), dolphin morbillivirus (DMV) and pilot whale morbillivirus (PWMV). Relative sensitivity was measured by comparing the results obtained from 10-fold dilution series of PMV and DMV positive controls and a PWMV field sample, to those obtained by the previously described conventional phosphoprotein gene based RT-PCR method. Both the conventional and real-time RT-PCR methods involving the fusion protein gene were 100- to 1000-fold more sensitive than the previously described conventional RT-PCR method. Copyright © 2015 Elsevier B.V. All rights reserved.

Crypto-Giardia antigen rapid test versus conventional modified Ziehl-Neelsen acid fast staining method for diagnosis of cryptosporidiosis.

PubMed

Zaglool, Dina Abdulla Muhammad; Mohamed, Amr; Khodari, Yousif Abdul Wahid; Farooq, Mian Usman

2013-03-01

To evaluate the validity of Crypto-Giardia antigen rapid test (CA-RT) in comparison with the conventional modified Ziehl-Neelsen acid fast (MZN-AF) staining method for the diagnosis of cryptosporidiosis. Fifteen preserved stool samples from previously confirmed infections were used as positive controls and 40 stool samples from healthy people were used as negative control. A total of 85 stool samples were collected from suspected patients with cryptosporidiosis over 6 months during the period from January till June, 2011. The study was conducted in the department of parasitology, central laboratory, Alnoor Specialist Hospital, Makkah, Saudi Arabia. All samples were subjected to CA-RT and conventional MZN-AF staining method. Validation parameters including sensitivity (SN), specificity (SP), accuracy index (AI), positive predictive value (PPV), and negative predictive value (NPV) were evaluated for both tests. Out of 15 positive controls, CA-RT detected 13 (86.7%) while MZN-AF detected 11(73.3%) positive cases. However, CA-RT detected no positive case in 40 normal controls but MZN-AF detected 2(5%) as positive cases. Based on the results, the SN, SP, AI, PPV and NPV were high in CA-RT than MZN-AF staining method, ie., 86.7%vs. 73.3%, 100%vs. 95%, 96.4%vs. 89.1%, 100%vs. 84.6% and 95.2%vs. 90.5%, respectively. Out of a total of 85 suspected specimens, CA-RT detected 7(8.2%) but MZN-AF detected 6(7.1%) cases as positive. CA-RT immunoassay is more valid and reliable than MZN-AF staining method. Copyright © 2013 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Comparison between laser terahertz emission microscope and conventional methods for analysis of polycrystalline silicon solar cell

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

Nakanishi, Hidetoshi, E-mail: nakanisi@screen.co.jp; Ito, Akira, E-mail: a.ito@screen.co.jp; Takayama, Kazuhisa, E-mail: takayama.k0123@gmail.com

2015-11-15

A laser terahertz emission microscope (LTEM) can be used for noncontact inspection to detect the waveforms of photoinduced terahertz emissions from material devices. In this study, we experimentally compared the performance of LTEM with conventional analysis methods, e.g., electroluminescence (EL), photoluminescence (PL), and laser beam induced current (LBIC), as an inspection method for solar cells. The results showed that LTEM was more sensitive to the characteristics of the depletion layer of the polycrystalline solar cell compared with EL, PL, and LBIC and that it could be used as a complementary tool to the conventional analysis methods for a solar cell.

Selective Detection of Peptide-Oligonucleotide Heteroconjugates Utilizing Capillary HPLC-ICPMS

NASA Astrophysics Data System (ADS)

Catron, Brittany; Caruso, Joseph A.; Limbach, Patrick A.

2012-06-01

A method for the selective detection and quantification of peptide:oligonucleotide heteroconjugates, such as those generated by protein:nucleic acid cross-links, using capillary reversed-phase high performance liquid chromatography (cap-RPHPLC) coupled with inductively coupled plasma mass spectrometry detection (ICPMS) is described. The selective detection of phosphorus as 31P+, the only natural isotope, in peptide-oligonucleotide heteroconjugates is enabled by the elemental detection capabilities of the ICPMS. Mobile phase conditions that allow separation of heteroconjugates while maintaining ICPMS compatibility were investigated. We found that trifluoroacetic acid (TFA) mobile phases, used in conventional peptide separations, and hexafluoroisopropanol/triethylamine (HFIP/TEA) mobile phases, used in conventional oligonucleotide separations, both are compatible with ICPMS and enable heteroconjugate separation. The TFA-based separations yielded limits of detection (LOD) of ~40 ppb phosphorus, which is nearly seven times lower than the LOD for HFIP/TEA-based separations. Using the TFA mobile phase, 1-2 pmol of a model heteroconjugate were routinely separated and detected by this optimized capLC-ICPMS method.

Modified short-term guinea pig sensitization tests for detecting contact allergens as an alternative to the conventional test.

PubMed

Yanagi, M; Hoya, M; Mori, M; Katsumura, Y

2001-03-01

The conventional adjuvant and patch test (APT) method of guinea pig sensitization testing was modified in 2 ways, s-APT and s-APT(2), in order to shorten the test period. These short-term test methods consist of 72-h closed application of test material with intradermal injection of emulsified Freund's complete adjuvant (E-FCA) for 1st induction, 48-h closed application of test material with (s-APT) or without (s-APT(2)) intradermal injection of E-FCA on the 7th day for 2nd induction, and open application on the 14th day for challenge. They were compared with conventional APT by using 8 allergenic chemicals (formaldehyde, nickel sulfate, cobalt sulfate, ethyl-p-aminobenzoate (benzocaine), isoeugenol, 2-mercaptobenzothiazole, 2,4-dinitrochlorobenzene (DNCB) and 1-phenylazo-2-naphthol (Sudan I)). The short-term methods gave similar results to those of conventional APT in terms of mean response, sensitization rate and sensitization potency (challenge concentration that induces a mean response equal to 1.0). Thus, our short-term methods, which are capable of evaluating skin sensitization within 17 days, are sufficiently sensitive to detect potentially hazardous contact allergens.

Habitat complexity and fish size affect the detection of Indo-Pacific lionfish on invaded coral reefs

NASA Astrophysics Data System (ADS)

Green, S. J.; Tamburello, N.; Miller, S. E.; Akins, J. L.; Côté, I. M.

2013-06-01

A standard approach to improving the accuracy of reef fish population estimates derived from underwater visual censuses (UVCs) is the application of species-specific correction factors, which assumes that a species' detectability is constant under all conditions. To test this assumption, we quantified detection rates for invasive Indo-Pacific lionfish ( Pterois volitans and P. miles), which are now a primary threat to coral reef conservation throughout the Caribbean. Estimates of lionfish population density and distribution, which are essential for managing the invasion, are currently obtained through standard UVCs. Using two conventional UVC methods, the belt transect and stationary visual census (SVC), we assessed how lionfish detection rates vary with lionfish body size and habitat complexity (measured as rugosity) on invaded continuous and patch reefs off Cape Eleuthera, the Bahamas. Belt transect and SVC surveys performed equally poorly, with both methods failing to detect the presence of lionfish in >50 % of surveys where thorough, lionfish-focussed searches yielded one or more individuals. Conventional methods underestimated lionfish biomass by ~200 %. Crucially, detection rate varied significantly with both lionfish size and reef rugosity, indicating that the application of a single correction factor across habitats and stages of invasion is unlikely to accurately characterize local populations. Applying variable correction factors that account for site-specific lionfish size and rugosity to conventional survey data increased estimates of lionfish biomass, but these remained significantly lower than actual biomass. To increase the accuracy and reliability of estimates of lionfish density and distribution, monitoring programs should use detailed area searches rather than standard visual survey methods. Our study highlights the importance of accounting for sources of spatial and temporal variation in detection to increase the accuracy of survey data from coral reef systems.

Comparison of the Microbiological Quality and Safety between Conventional and Organic Vegetables Sold in Malaysia

PubMed Central

Kuan, Chee-Hao; Rukayadi, Yaya; Ahmad, Siti H.; Wan Mohamed Radzi, Che W. J.; Thung, Tze-Young; Premarathne, Jayasekara M. K. J. K.; Chang, Wei-San; Loo, Yuet-Ying; Tan, Chia-Wanq; Ramzi, Othman B.; Mohd Fadzil, Siti N.; Kuan, Chee-Sian; Yeo, Siok-Koon; Nishibuchi, Mitsuaki; Radu, Son

2017-01-01

Given the remarkable increase of public interest in organic food products, it is indeed critical to evaluate the microbiological risk associated with consumption of fresh organic produce. Organic farming practices including the use of animal manures may increase the risk of microbiological contamination as manure can act as a vehicle for transmission of foodborne pathogens. This study aimed to determine and compare the microbiological status between organic and conventional fresh produce at the retail level in Malaysia. A total of 152 organic and conventional vegetables were purchased at retail markets in Malaysia. Samples were analyzed for mesophilic aerobic bacteria, yeasts and molds, and total coliforms using conventional microbiological methods. Combination methods of most probable number-multiplex polymerase chain reaction (MPN-mPCR) were used to detect and quantify foodborne pathogens, including Escherichia coli O157:H7, Shiga toxin-producing E. coli (STEC), Listeria monocytogenes, Salmonella Typhimurium, and Salmonella Enteritidis. Results indicated that most types of organic and conventional vegetables possessed similar microbial count (P > 0.05) of mesophilic aerobic bacteria, yeasts and molds, and total coliforms. E. coli O157:H7 and S. Typhimurium were not detected in any sample analyzed in this study. Among the 152 samples tested, only the conventional lettuce and organic carrot were tested positive for STEC and S. Enteritidis, respectively. L. monocytogenes were more frequently detected in both organic (9.1%) and conventional vegetables (2.7%) as compared to E. coli O157:H7, S. Typhimurium, and S. Enteritidis. Overall, no trend was shown that either organically or conventionally grown vegetables have posed greater microbiological risks. These findings indicated that one particular type of farming practices would not affect the microbiological profiles of fresh produce. Therefore, regardless of farming methods, all vegetables should be subjected to appropriate post-harvest handling practices from farm to fork to ensure the quality and safety of the fresh produce. PMID:28824567

Comparison of the Microbiological Quality and Safety between Conventional and Organic Vegetables Sold in Malaysia.

PubMed

Kuan, Chee-Hao; Rukayadi, Yaya; Ahmad, Siti H; Wan Mohamed Radzi, Che W J; Thung, Tze-Young; Premarathne, Jayasekara M K J K; Chang, Wei-San; Loo, Yuet-Ying; Tan, Chia-Wanq; Ramzi, Othman B; Mohd Fadzil, Siti N; Kuan, Chee-Sian; Yeo, Siok-Koon; Nishibuchi, Mitsuaki; Radu, Son

2017-01-01

Given the remarkable increase of public interest in organic food products, it is indeed critical to evaluate the microbiological risk associated with consumption of fresh organic produce. Organic farming practices including the use of animal manures may increase the risk of microbiological contamination as manure can act as a vehicle for transmission of foodborne pathogens. This study aimed to determine and compare the microbiological status between organic and conventional fresh produce at the retail level in Malaysia. A total of 152 organic and conventional vegetables were purchased at retail markets in Malaysia. Samples were analyzed for mesophilic aerobic bacteria, yeasts and molds, and total coliforms using conventional microbiological methods. Combination methods of most probable number-multiplex polymerase chain reaction (MPN-mPCR) were used to detect and quantify foodborne pathogens, including Escherichia coli O157:H7, Shiga toxin-producing E. coli (STEC), Listeria monocytogenes, Salmonella Typhimurium, and Salmonella Enteritidis. Results indicated that most types of organic and conventional vegetables possessed similar microbial count ( P > 0.05) of mesophilic aerobic bacteria, yeasts and molds, and total coliforms. E. coli O157:H7 and S . Typhimurium were not detected in any sample analyzed in this study. Among the 152 samples tested, only the conventional lettuce and organic carrot were tested positive for STEC and S . Enteritidis, respectively. L. monocytogenes were more frequently detected in both organic (9.1%) and conventional vegetables (2.7%) as compared to E. coli O157:H7, S . Typhimurium, and S . Enteritidis. Overall, no trend was shown that either organically or conventionally grown vegetables have posed greater microbiological risks. These findings indicated that one particular type of farming practices would not affect the microbiological profiles of fresh produce. Therefore, regardless of farming methods, all vegetables should be subjected to appropriate post-harvest handling practices from farm to fork to ensure the quality and safety of the fresh produce.

Label-free detection of salmonella typhimurium with ssDNA aptamers

USDA-ARS?s Scientific Manuscript database

Foodborne pathogen Salmonella enterica is one of the major causes of gastrointestinal infections in human and animals. Conventional detection methods are time consuming and not effective enough under emergency circumstances to control outbreaks immediately. Therefore, biosensors that can detect Salm...

Optimal weighting of data to detect climatic change - Application to the carbon dioxide problem

NASA Technical Reports Server (NTRS)

Bell, T. L.

1982-01-01

It is suggested that a weighting of surface temperature data, using information about the expected level of warming in different seasons and geographical regions and statistical information about the amount of natural variability in surface temperature, can improve the chances of early detection of carbon dioxide concentration-induced climatic warming. A preliminary analysis of the optimal weighting method presented suggests that it is 25 per cent more effective in revealing surface warming than the conventional method, in virtue of the fact that 25 per cent more data must conventionally be analyzed in order to arrive at a similar probability of detection. An approximate calculation suggests that the warming ought to have already been detected, if the only sources of significant surface temperature variability had time scales of less than one year.

Adaptive skin detection based on online training

NASA Astrophysics Data System (ADS)

Zhang, Ming; Tang, Liang; Zhou, Jie; Rong, Gang

2007-11-01

Skin is a widely used cue for porn image classification. Most conventional methods are off-line training schemes. They usually use a fixed boundary to segment skin regions in the images and are effective only in restricted conditions: e.g. good lightness and unique human race. This paper presents an adaptive online training scheme for skin detection which can handle these tough cases. In our approach, skin detection is considered as a classification problem on Gaussian mixture model. For each image, human face is detected and the face color is used to establish a primary estimation of skin color distribution. Then an adaptive online training algorithm is used to find the real boundary between skin color and background color in current image. Experimental results on 450 images showed that the proposed method is more robust in general situations than the conventional ones.

Conventional and phase contrast x-ray imaging techniques and ultrasound imaging method in breast tumor detection: initial comparison studies using phantom

NASA Astrophysics Data System (ADS)

Guo, Yuran; Wu, Di; Omoumi, Farid H.; Li, Yuhua; Wong, Molly Donovan; Ghani, Muhammad U.; Zheng, Bin; Liu, Hong

2018-02-01

The objective of this study was to demonstrate the capability of the high-energy in-line phase contrast imaging in detecting the breast tumors which are undetectable by conventional x-ray imaging but detectable by ultrasound. Experimentally, a CIRS multipurpose breast phantom with heterogeneous 50% glandular and 50% adipose breast tissue was imaged by high-energy in-line phase contrast system, conventional x-ray system and ultrasonography machine. The high-energy in-line phase contrast projection was acquired at 120 kVp, 0.3 mAs with the focal spot size of 18.3 μm. The conventional x-ray projection was acquired at 40 kVp, 3.3 mAs with the focal spot size of 22.26 μm. Both of the x-ray imaging acquisitions were conducted with a unique mean glandular dose of 0.08 mGy. As the result, the high-energy in-line phase contrast system was able to detect one lesion-like object which was also detected by the ultrasonography. This object was spherical shape with the length of about 12.28 mm. Also, the conventional x-ray system was not able to detect any objects. This result indicated the advantages provided by high-energy in-line phase contrast over conventional x-ray system in detecting lesion-like object under the same radiation dose. To meet the needs of current clinical strategies for high-density breasts screening, breast phantoms with higher glandular densities will be employed in future studies.

An appropriately performed conventional blood culture can facilitate choice of therapy in resource-constrained settings-comparison with BACTEC 9050.

PubMed

Surase, P V; Nataraj, G; Pattamadai, K; Mehta, P R; Pazare, A R; Agarwal, M C; Nanavati, R N

2016-01-01

Comparison of conventional blood culture with BACTEC 9050 for rate and time to detection of microorganisms. A prospective study was carried out in a multispecialty tertiary care teaching hospital. A total of 835 paired specimens (797 blood and 38 nonblood specimens) were collected and processed according to standard microbiological procedures by both conventional method as well as by BACTEC 9050 automated culture system. Clinical details of patients were recorded. Data were analyzed for time to detection and isolation rate by the two systems and compared. Overall culture positivity for BACTEC 9050 and the conventional system was 32% and 19.88%, respectively. Eighty-five demonstrated concordant growth, 136 specimens were culture positive by BACTEC only, and 38 specimens were culture positive by conventional only. Twelve contaminants in BACTEC and nine contaminants in conventional system were detected. Using BACTEC 9050, higher isolation was observed for Acinetobacter spp., coagulase negative Staphylococcus spp., Streptococcus spp., and Candida spp. A total of 410 patients were on antimicrobial treatment and culture positivity was significantly higher with BACTEC 9050 (P < 0.0001). There was a significant difference in the mean time to detection with BACTEC 9050 recovering 86.8% of isolates within 48 h (P < 0.0001). Although BACTEC 9050 demonstrated a significantly higher recovery of microorganisms from blood, an appropriately performed conventional blood culture can facilitate the choice of therapy.

Comparison of the microbiological quality of environmentally friendly and conventionally grown vegetables sold at retail markets in Korea.

PubMed

Ryu, Jee-Hoon; Kim, Minju; Kim, Eun-Gyeong; Beuchat, Larry R; Kim, Hoikyung

2014-09-01

Fresh produce is usually eaten raw without cooking or heating, which may increase the probability of foodborne infection. The microbiological quality of 11 types of fresh, raw vegetables (romaine lettuce, sesame leaves, crown daisy, garlic chives, iceberg lettuce, cabbage, broccoli, leek, chili pepper, capsicum, and zucchini) purchased at retail markets in Iksan, Korea as affected by cultivation method (environmentally friendly vegetables [organic, pesticide-free, and low-pesticide vegetables] and conventionally grown vegetables) and harvest season was determined. Escherichia coli O157:H7 and Salmonella were not detected in all samples of vegetables tested. Aerobic mesophiles (>6 log cfu/g) were detected in environmentally friendly romaine lettuce and crown daisy and environmentally friendly and conventionally grown garlic chives, which also contained coliforms (>3 log cfu/g). Sesame leaf and crown daisy (regardless of cultivation method), as well as conventionally grown romaine lettuce and leek, contained >1 log cfu/g of E. coli. The overall microbiological quality of environmentally friendly and conventionally grown vegetables was not significantly different (P > 0.05). However, there were seasonal effects on populations of coliforms and generic E. coli on vegetables. The greatest numbers of microorganisms were isolated from environmentally friendly or conventionally grown vegetables purchased in winter. The vegetables, regardless of cultivation method or season, should be subjected to appropriate antimicrobial treatment to enhance their microbial safety. © 2014 Institute of Food Technologists®

Advances in Candida detection platforms for clinical and point-of-care applications

PubMed Central

Safavieh, Mohammadali; Coarsey, Chad; Esiobu, Nwadiuto; Memic, Adnan; Vyas, Jatin Mahesh; Shafiee, Hadi; Asghar, Waseem

2016-01-01

Invasive candidiasis remains one of the most serious community and healthcare-acquired infections worldwide. Conventional Candida detection methods based on blood and plate culture are time-consuming and require at least 2–4 days to identify various Candida species. Despite considerable advances for candidiasis detection, the development of simple, compact and portable point-of-care diagnostics for rapid and precise testing that automatically performs cell lysis, nucleic acid extraction, purification and detection still remains a challenge. Here, we systematically review most prominent conventional and nonconventional techniques for the detection of various Candida species, including Candida staining, blood culture, serological testing and nucleic acid-based analysis. We also discuss the most advanced lab on a chip devices for candida detection. PMID:27093473

An automated cross-correlation based event detection technique and its application to surface passive data set

USGS Publications Warehouse

Forghani-Arani, Farnoush; Behura, Jyoti; Haines, Seth S.; Batzle, Mike

2013-01-01

In studies on heavy oil, shale reservoirs, tight gas and enhanced geothermal systems, the use of surface passive seismic data to monitor induced microseismicity due to the fluid flow in the subsurface is becoming more common. However, in most studies passive seismic records contain days and months of data and manually analysing the data can be expensive and inaccurate. Moreover, in the presence of noise, detecting the arrival of weak microseismic events becomes challenging. Hence, the use of an automated, accurate and computationally fast technique for event detection in passive seismic data is essential. The conventional automatic event identification algorithm computes a running-window energy ratio of the short-term average to the long-term average of the passive seismic data for each trace. We show that for the common case of a low signal-to-noise ratio in surface passive records, the conventional method is not sufficiently effective at event identification. Here, we extend the conventional algorithm by introducing a technique that is based on the cross-correlation of the energy ratios computed by the conventional method. With our technique we can measure the similarities amongst the computed energy ratios at different traces. Our approach is successful at improving the detectability of events with a low signal-to-noise ratio that are not detectable with the conventional algorithm. Also, our algorithm has the advantage to identify if an event is common to all stations (a regional event) or to a limited number of stations (a local event). We provide examples of applying our technique to synthetic data and a field surface passive data set recorded at a geothermal site.

Sensor assembly method using silicon interposer with trenches for three-dimensional binocular range sensors

NASA Astrophysics Data System (ADS)

Nakajima, Kazuhiro; Yamamoto, Yuji; Arima, Yutaka

2018-04-01

To easily assemble a three-dimensional binocular range sensor, we devised an alignment method for two image sensors using a silicon interposer with trenches. The trenches were formed using deep reactive ion etching (RIE) equipment. We produced a three-dimensional (3D) range sensor using the method and experimentally confirmed that sufficient alignment accuracy was realized. It was confirmed that the alignment accuracy of the two image sensors when using the proposed method is more than twice that of the alignment assembly method on a conventional board. In addition, as a result of evaluating the deterioration of the detection performance caused by the alignment accuracy, it was confirmed that the vertical deviation between the corresponding pixels in the two image sensors is substantially proportional to the decrease in detection performance. Therefore, we confirmed that the proposed method can realize more than twice the detection performance of the conventional method. Through these evaluations, the effectiveness of the 3D binocular range sensor aligned by the silicon interposer with the trenches was confirmed.

Differentiation of live and dead salmonella cells using fourier transform infrared (FTIR) spectroscopy and principle component analysis (PCA) technique

USDA-ARS?s Scientific Manuscript database

Various technologies have been developed for pathogen detection using optical, electrochemical, biochemical and physical properties. Conventional microbiological methods need time from days to week to get the result. Though this method is very sensitive and accurate, a rapid detection of pathogens i...

Current trends in endotoxin detection and analysis of endotoxin-protein interactions.

PubMed

Dullah, Elvina Clarie; Ongkudon, Clarence M

2017-03-01

Endotoxin is a type of pyrogen that can be found in Gram-negative bacteria. Endotoxin can form a stable interaction with other biomolecules thus making its removal difficult especially during the production of biopharmaceutical drugs. The prevention of endotoxins from contaminating biopharmaceutical products is paramount as endotoxin contamination, even in small quantities, can result in fever, inflammation, sepsis, tissue damage and even lead to death. Highly sensitive and accurate detection of endotoxins are keys in the development of biopharmaceutical products derived from Gram-negative bacteria. It will facilitate the study of the intermolecular interaction of an endotoxin with other biomolecules, hence the selection of appropriate endotoxin removal strategies. Currently, most researchers rely on the conventional LAL-based endotoxin detection method. However, new methods have been and are being developed to overcome the problems associated with the LAL-based method. This review paper highlights the current research trends in endotoxin detection from conventional methods to newly developed biosensors. Additionally, it also provides an overview of the use of electron microscopy, dynamic light scattering (DLS), fluorescence resonance energy transfer (FRET) and docking programs in the endotoxin-protein analysis.

Screening for Human Immunodeficiency Virus, Hepatitis B Virus, Hepatitis C Virus, and Treponema pallidum by Blood Testing Using a Bio-Flash Technology-Based Algorithm before Gastrointestinal Endoscopy

PubMed Central

Zhen, Chen; QuiuLi, Zhang; YuanQi, An; Casado, Verónica Vocero; Fan, Yuan

2016-01-01

Currently, conventional enzyme immunoassays which use manual gold immunoassays and colloidal tests (GICTs) are used as screening tools to detect Treponema pallidum (syphilis), hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus type 1 (HIV-1), and HIV-2 in patients undergoing surgery. The present observational, cross-sectional study compared the sensitivity, specificity, and work flow characteristics of the conventional algorithm with manual GICTs with those of a newly proposed algorithm that uses the automated Bio-Flash technology as a screening tool in patients undergoing gastrointestinal (GI) endoscopy. A total of 956 patients were examined for the presence of serological markers of infection with HIV-1/2, HCV, HBV, and T. pallidum. The proposed algorithm with the Bio-Flash technology was superior for the detection of all markers (100.0% sensitivity and specificity for detection of anti-HIV and anti-HCV antibodies, HBV surface antigen [HBsAg], and T. pallidum) compared with the conventional algorithm based on the manual method (80.0% sensitivity and 98.6% specificity for the detection of anti-HIV, 75.0% sensitivity for the detection of anti-HCV, 94.7% sensitivity for the detection of HBsAg, and 100% specificity for the detection of anti-HCV and HBsAg) in these patients. The automated Bio-Flash technology-based screening algorithm also reduced the operation time by 85.0% (205 min) per day, saving up to 24 h/week. In conclusion, the use of the newly proposed screening algorithm based on the automated Bio-Flash technology can provide an advantage over the use of conventional algorithms based on manual methods for screening for HIV, HBV, HCV, and syphilis before GI endoscopy. PMID:27707942

Screening for Human Immunodeficiency Virus, Hepatitis B Virus, Hepatitis C Virus, and Treponema pallidum by Blood Testing Using a Bio-Flash Technology-Based Algorithm before Gastrointestinal Endoscopy.

PubMed

Jun, Zhou; Zhen, Chen; QuiuLi, Zhang; YuanQi, An; Casado, Verónica Vocero; Fan, Yuan

2016-12-01

Currently, conventional enzyme immunoassays which use manual gold immunoassays and colloidal tests (GICTs) are used as screening tools to detect Treponema pallidum (syphilis), hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus type 1 (HIV-1), and HIV-2 in patients undergoing surgery. The present observational, cross-sectional study compared the sensitivity, specificity, and work flow characteristics of the conventional algorithm with manual GICTs with those of a newly proposed algorithm that uses the automated Bio-Flash technology as a screening tool in patients undergoing gastrointestinal (GI) endoscopy. A total of 956 patients were examined for the presence of serological markers of infection with HIV-1/2, HCV, HBV, and T. pallidum The proposed algorithm with the Bio-Flash technology was superior for the detection of all markers (100.0% sensitivity and specificity for detection of anti-HIV and anti-HCV antibodies, HBV surface antigen [HBsAg], and T. pallidum) compared with the conventional algorithm based on the manual method (80.0% sensitivity and 98.6% specificity for the detection of anti-HIV, 75.0% sensitivity for the detection of anti-HCV, 94.7% sensitivity for the detection of HBsAg, and 100% specificity for the detection of anti-HCV and HBsAg) in these patients. The automated Bio-Flash technology-based screening algorithm also reduced the operation time by 85.0% (205 min) per day, saving up to 24 h/week. In conclusion, the use of the newly proposed screening algorithm based on the automated Bio-Flash technology can provide an advantage over the use of conventional algorithms based on manual methods for screening for HIV, HBV, HCV, and syphilis before GI endoscopy. Copyright © 2016 Jun et al.

Covariance descriptor fusion for target detection

NASA Astrophysics Data System (ADS)

Cukur, Huseyin; Binol, Hamidullah; Bal, Abdullah; Yavuz, Fatih

2016-05-01

Target detection is one of the most important topics for military or civilian applications. In order to address such detection tasks, hyperspectral imaging sensors provide useful images data containing both spatial and spectral information. Target detection has various challenging scenarios for hyperspectral images. To overcome these challenges, covariance descriptor presents many advantages. Detection capability of the conventional covariance descriptor technique can be improved by fusion methods. In this paper, hyperspectral bands are clustered according to inter-bands correlation. Target detection is then realized by fusion of covariance descriptor results based on the band clusters. The proposed combination technique is denoted Covariance Descriptor Fusion (CDF). The efficiency of the CDF is evaluated by applying to hyperspectral imagery to detect man-made objects. The obtained results show that the CDF presents better performance than the conventional covariance descriptor.

Detecting the sampling rate through observations

NASA Astrophysics Data System (ADS)

Shoji, Isao

2018-09-01

This paper proposes a method to detect the sampling rate of discrete time series of diffusion processes. Using the maximum likelihood estimates of the parameters of a diffusion process, we establish a criterion based on the Kullback-Leibler divergence and thereby estimate the sampling rate. Simulation studies are conducted to check whether the method can detect the sampling rates from data and their results show a good performance in the detection. In addition, the method is applied to a financial time series sampled on daily basis and shows the detected sampling rate is different from the conventional rates.

Comparison of Real-Time PCR, Reverse Transcriptase Real-Time PCR, Loop-Mediated Isothermal Amplification, and the FDA Conventional Microbiological Method for the Detection of Salmonella spp. in Produce ▿ †

PubMed Central

Zhang, Guodong; Brown, Eric W.; González-Escalona, Narjol

2011-01-01

Contamination of foods, especially produce, with Salmonella spp. is a major concern for public health. Several methods are available for the detection of Salmonella in produce, but their relative efficiency for detecting Salmonella in commonly consumed vegetables, often associated with outbreaks of food poisoning, needs to be confirmed. In this study, the effectiveness of three molecular methods for detection of Salmonella in six produce matrices was evaluated and compared to the FDA microbiological detection method. Samples of cilantro (coriander leaves), lettuce, parsley, spinach, tomato, and jalapeno pepper were inoculated with Salmonella serovars at two different levels (105 and <101 CFU/25 g of produce). The inoculated produce was assayed by the FDA Salmonella culture method (Bacteriological Analytical Manual) and by three molecular methods: quantitative real-time PCR (qPCR), quantitative reverse transcriptase real-time PCR (RT-qPCR), and loop-mediated isothermal amplification (LAMP). Comparable results were obtained by these four methods, which all detected as little as 2 CFU of Salmonella cells/25 g of produce. All control samples (not inoculated) were negative by the four methods. RT-qPCR detects only live Salmonella cells, obviating the danger of false-positive results from nonviable cells. False negatives (inhibition of either qPCR or RT-qPCR) were avoided by the use of either a DNA or an RNA amplification internal control (IAC). Compared to the conventional culture method, the qPCR, RT-qPCR, and LAMP assays allowed faster and equally accurate detection of Salmonella spp. in six high-risk produce commodities. PMID:21803916

Single-tube analysis of DNA methylation with silica superparamagnetic beads.

PubMed

Bailey, Vasudev J; Zhang, Yi; Keeley, Brian P; Yin, Chao; Pelosky, Kristen L; Brock, Malcolm; Baylin, Stephen B; Herman, James G; Wang, Tza-Huei

2010-06-01

DNA promoter methylation is a signature for the silencing of tumor suppressor genes. Most widely used methods to detect DNA methylation involve 3 separate, independent processes: DNA extraction, bisulfite conversion, and methylation detection via a PCR method, such as methylation-specific PCR (MSP). This method includes many disconnected steps with associated losses of material, potentially reducing the analytical sensitivity required for analysis of challenging clinical samples. Methylation on beads (MOB) is a new technique that integrates DNA extraction, bisulfite conversion, and PCR in a single tube via the use of silica superparamagnetic beads (SSBs) as a common DNA carrier for facilitating cell debris removal and buffer exchange throughout the entire process. In addition, PCR buffer is used to directly elute bisulfite-treated DNA from SSBs for subsequent target amplifications. The diagnostic sensitivity of MOB was evaluated by methylation analysis of the CDKN2A [cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4); also known as p16(INK4a)] promoter in serum DNA of lung cancer patients and compared with that of conventional methods. Methylation analysis consisting of DNA extraction followed by bisulfite conversion and MSP was successfully carried out within 9 h in a single tube. The median pre-PCR DNA yield was 6.61-fold higher with the MOB technique than with conventional techniques. Furthermore, MOB increased the diagnostic sensitivity in our analysis of the CDKN2A promoter in patient serum by successfully detecting methylation in 74% of cancer patients, vs the 45% detection rate obtained with conventional techniques. The MOB technique successfully combined 3 processes into a single tube, thereby allowing ease in handling and an increased detection throughput. The increased pre-PCR yield in MOB allowed efficient, diagnostically sensitive methylation detection.

Monitoring the enzymatic conversion of urea to ammonium by conventional or microchip capillary electrophoresis with contactless conductivity detection.

PubMed

Schuchert-Shi, Aiping; Hauser, Peter C

2008-05-15

Capillary electrophoresis with contactless conductivity detection was used to directly quantify the ammonium produced in the enzymatic conversion of urea with urease. This allowed the characterization of the reaction without having to use more elaborate indirect optical methods for quantification. The maximum rate of reaction, V(max), was determined as 5.1 mmol x mL(-1) x min(-1), and the Michaelis-Menten constant, K(m), was determined as 16 mM. Furthermore, the method was successfully applied to the determination of urea in clinical samples of human blood by using a conventional capillary and a microchip device.

GammaScorpion: mobile gamma-ray tomography system for early detection of basal stem rot in oil palm plantations

NASA Astrophysics Data System (ADS)

Abdullah, Jaafar; Hassan, Hearie; Shari, Mohamad Rabaie; Mohd, Salzali; Mustapha, Mahadi; Mahmood, Airwan Affendi; Jamaludin, Shahrizan; Ngah, Mohd Rosdi; Hamid, Noor Hisham

2013-03-01

Detection of the oil palm stem rot disease Ganoderma is a major issue in estate management and production in Malaysia. Conventional diagnostic techniques are difficult and time consuming when using visual inspection, and destructive and expensive when based on the chemical analysis of root or stem tissue. As an alternative, a transportable gamma-ray computed tomography system for the early detection of basal stem rot (BSR) of oil palms due to Ganoderma was developed locally at the Malaysian Nuclear Agency, Kajang, Malaysia. This system produces high quality tomographic images that clearly differentiate between healthy and Ganoderma infected oil palm stems. It has been successfully tested and used to detect the extent of BSR damage in oil palm plantations in Malaysia without the need to cut down the trees. This method offers promise for in situ inspection of oil palm stem diseases compared to the more conventional methods.

Fetal head detection and measurement in ultrasound images by an iterative randomized Hough transform

NASA Astrophysics Data System (ADS)

Lu, Wei; Tan, Jinglu; Floyd, Randall C.

2004-05-01

This paper describes an automatic method for measuring the biparietal diameter (BPD) and head circumference (HC) in ultrasound fetal images. A total of 217 ultrasound images were segmented by using a K-Mean classifier, and the head skull was detected in 214 of the 217 cases by an iterative randomized Hough transform developed for detection of incomplete curves in images with strong noise without user intervention. The automatic measurements were compared with conventional manual measurements by sonographers and a trained panel. The inter-run variations and differences between the automatic and conventional measurements were small compared with published inter-observer variations. The results showed that the automated measurements were as reliable as the expert measurements and more consistent. This method has great potential in clinical applications.

Cost and detection rate of glaucoma screening with imaging devices in a primary care center

PubMed Central

Anton, Alfonso; Fallon, Monica; Cots, Francesc; Sebastian, María A; Morilla-Grasa, Antonio; Mojal, Sergi; Castells, Xavier

2017-01-01

Purpose To analyze the cost and detection rate of a screening program for detecting glaucoma with imaging devices. Materials and methods In this cross-sectional study, a glaucoma screening program was applied in a population-based sample randomly selected from a population of 23,527. Screening targeted the population at risk of glaucoma. Examinations included optic disk tomography (Heidelberg retina tomograph [HRT]), nerve fiber analysis, and tonometry. Subjects who met at least 2 of 3 endpoints (HRT outside normal limits, nerve fiber index ≥30, or tonometry ≥21 mmHg) were referred for glaucoma consultation. The currently established (“conventional”) detection method was evaluated by recording data from primary care and ophthalmic consultations in the same population. The direct costs of screening and conventional detection were calculated by adding the unit costs generated during the diagnostic process. The detection rate of new glaucoma cases was assessed. Results The screening program evaluated 414 subjects; 32 cases were referred for glaucoma consultation, 7 had glaucoma, and 10 had probable glaucoma. The current detection method assessed 677 glaucoma suspects in the population, of whom 29 were diagnosed with glaucoma or probable glaucoma. Glaucoma screening and the conventional detection method had detection rates of 4.1% and 3.1%, respectively, and the cost per case detected was 1,410 and 1,435€, respectively. The cost of screening 1 million inhabitants would be 5.1 million euros and would allow the detection of 4,715 new cases. Conclusion The proposed screening method directed at population at risk allows a detection rate of 4.1% and a cost of 1,410 per case detected. PMID:28243057

How to detect carbapenemase producers? A literature review of phenotypic and molecular methods.

PubMed

Hammoudi, D; Moubareck, C Ayoub; Sarkis, D Karam

2014-12-01

This review describes the current state-of-art of carbapenemase detection methods. Identification of carbapenemases is first based on conventional phenotypic tests including antimicrobial susceptibility testing, modified-Hodge test and carbapenemase-inhibitor culture tests. Second, molecular characterization of carbapenemase genes by PCR sequencing is essential. Third, innovative biochemical and spectrometric detection may be applied. Copyright © 2014 Elsevier B.V. All rights reserved.

Comparison of the lysis centrifugation method with the conventional blood culture method in cases of sepsis in a tertiary care hospital.

PubMed

Parikh, Harshal R; De, Anuradha S; Baveja, Sujata M

2012-07-01

Physicians and microbiologists have long recognized that the presence of living microorganisms in the blood of a patient carries with it considerable morbidity and mortality. Hence, blood cultures have become critically important and frequently performed test in clinical microbiology laboratories for diagnosis of sepsis. To compare the conventional blood culture method with the lysis centrifugation method in cases of sepsis. Two hundred nonduplicate blood cultures from cases of sepsis were analyzed using two blood culture methods concurrently for recovery of bacteria from patients diagnosed clinically with sepsis - the conventional blood culture method using trypticase soy broth and the lysis centrifugation method using saponin by centrifuging at 3000 g for 30 minutes. Overall bacteria recovered from 200 blood cultures were 17.5%. The conventional blood culture method had a higher yield of organisms, especially Gram positive cocci. The lysis centrifugation method was comparable with the former method with respect to Gram negative bacilli. The sensitivity of lysis centrifugation method in comparison to conventional blood culture method was 49.75% in this study, specificity was 98.21% and diagnostic accuracy was 89.5%. In almost every instance, the time required for detection of the growth was earlier by lysis centrifugation method, which was statistically significant. Contamination by lysis centrifugation was minimal, while that by conventional method was high. Time to growth by the lysis centrifugation method was highly significant (P value 0.000) as compared to time to growth by the conventional blood culture method. For the diagnosis of sepsis, combination of the lysis centrifugation method and the conventional blood culture method with trypticase soy broth or biphasic media is advocable, in order to achieve faster recovery and a better yield of microorganisms.

Comparison of four PCR methods for efficient detection of Trypanosoma cruzi in routine diagnostics.

PubMed

Seiringer, Peter; Pritsch, Michael; Flores-Chavez, María; Marchisio, Edoardo; Helfrich, Kerstin; Mengele, Carolin; Hohnerlein, Stefan; Bretzel, Gisela; Löscher, Thomas; Hoelscher, Michael; Berens-Riha, Nicole

2017-07-01

Due to increased migration, Chagas disease has become an international health problem. Reliable diagnosis of chronically infected people is crucial for prevention of non-vectorial transmission as well as treatment. This study compared four distinct PCR methods for detection of Trypanosoma cruzi DNA for the use in well-equipped routine diagnostic laboratories. DNA was extracted of T. cruzi-positive and negative patients' blood samples and cultured T. cruzi, T. rangeli as well as Leishmania spp. One conventional and two real-time PCR methods targeting a repetitive Sat-DNA sequence as well as one conventional PCR method targeting the variable region of the kDNA minicircle were compared for sensitivity, intra- and interassay precision, limit of detection, specificity and cross-reactivity. Considering the performance, costs and ease of use, an algorithm for PCR-diagnosis of patients with a positive serology for T. cruzi antibodies was developed. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

Bio-barcode gel assay for microRNA

NASA Astrophysics Data System (ADS)

Lee, Hyojin; Park, Jeong-Eun; Nam, Jwa-Min

2014-02-01

MicroRNA has been identified as a potential biomarker because expression level of microRNA is correlated with various cancers. Its detection at low concentrations would be highly beneficial for cancer diagnosis. Here, we develop a new type of a DNA-modified gold nanoparticle-based bio-barcode assay that uses a conventional gel electrophoresis platform and potassium cyanide chemistry and show this assay can detect microRNA at aM levels without enzymatic amplification. It is also shown that single-base-mismatched microRNA can be differentiated from perfectly matched microRNA and the multiplexed detection of various combinations of microRNA sequences is possible with this approach. Finally, differently expressed microRNA levels are selectively detected from cancer cells using the bio-barcode gel assay, and the results are compared with conventional polymerase chain reaction-based results. The method and results shown herein pave the way for practical use of a conventional gel electrophoresis for detecting biomolecules of interest even at aM level without polymerase chain reaction amplification.

Assessment of cerebral venous sinus thrombosis using T2*-weighted gradient echo magnetic resonance imaging sequences

PubMed Central

Bidar, Fatemeh; Faeghi, Fariborz; Ghorbani, Askar

2016-01-01

Background: The purpose of this study is to demonstrate the advantages of gradient echo (GRE) sequences in the detection and characterization of cerebral venous sinus thrombosis compared to conventional magnetic resonance sequences. Methods: A total of 17 patients with cerebral venous thrombosis (CVT) were evaluated using different magnetic resonance imaging (MRI) sequences. The MRI sequences included T1-weighted spin echo (SE) imaging, T*2-weighted turbo SE (TSE), fluid attenuated inversion recovery (FLAIR), T*2-weighted conventional GRE, and diffusion weighted imaging (DWI). MR venography (MRV) images were obtained as the golden standard. Results: Venous sinus thrombosis was best detectable in T*2-weighted conventional GRE sequences in all patients except in one case. Venous thrombosis was undetectable in DWI. T*2-weighted GRE sequences were superior to T*2-weighted TSE, T1-weighted SE, and FLAIR. Enhanced MRV was successful in displaying the location of thrombosis. Conclusion: T*2-weighted conventional GRE sequences are probably the best method for the assessment of cerebral venous sinus thrombosis. The mentioned method is non-invasive; therefore, it can be employed in the clinical evaluation of cerebral venous sinus thrombosis. PMID:27326365

Relative sensitivity of conventional and real-time PCR assays for detection of SFG Rickettsia in blood and tissue samples from laboratory animals.

PubMed

Zemtsova, Galina E; Montgomery, Merrill; Levin, Michael L

2015-01-01

Studies on the natural transmission cycles of zoonotic pathogens and the reservoir competence of vertebrate hosts require methods for reliable diagnosis of infection in wild and laboratory animals. Several PCR-based applications have been developed for detection of infections caused by Spotted Fever group Rickettsia spp. in a variety of animal tissues. These assays are being widely used by researchers, but they differ in their sensitivity and reliability. We compared the sensitivity of five previously published conventional PCR assays and one SYBR green-based real-time PCR assay for the detection of rickettsial DNA in blood and tissue samples from Rickettsia- infected laboratory animals (n = 87). The real-time PCR, which detected rickettsial DNA in 37.9% of samples, was the most sensitive. The next best were the semi-nested ompA assay and rpoB conventional PCR, which detected as positive 18.4% and 14.9% samples respectively. Conventional assays targeting ompB, gltA and hrtA genes have been the least sensitive. Therefore, we recommend the SYBR green-based real-time PCR as a tool for the detection of rickettsial DNA in animal samples due to its higher sensitivity when compared to more traditional assays.

Relative Sensitivity of Conventional and Real-Time PCR Assays for Detection of SFG Rickettsia in Blood and Tissue Samples from Laboratory Animals

PubMed Central

Zemtsova, Galina E.; Montgomery, Merrill; Levin, Michael L.

2015-01-01

Studies on the natural transmission cycles of zoonotic pathogens and the reservoir competence of vertebrate hosts require methods for reliable diagnosis of infection in wild and laboratory animals. Several PCR-based applications have been developed for detection of infections caused by Spotted Fever group Rickettsia spp. in a variety of animal tissues. These assays are being widely used by researchers, but they differ in their sensitivity and reliability. We compared the sensitivity of five previously published conventional PCR assays and one SYBR green-based real-time PCR assay for the detection of rickettsial DNA in blood and tissue samples from Rickettsia- infected laboratory animals (n = 87). The real-time PCR, which detected rickettsial DNA in 37.9% of samples, was the most sensitive. The next best were the semi-nested ompA assay and rpoB conventional PCR, which detected as positive 18.4% and 14.9% samples respectively. Conventional assays targeting ompB, gltA and hrtA genes have been the least sensitive. Therefore, we recommend the SYBR green-based real-time PCR as a tool for the detection of rickettsial DNA in animal samples due to its higher sensitivity when compared to more traditional assays. PMID:25607846

Raman Hyperspectral Imaging for Detection of Watermelon Seeds Infected with Acidovorax citrulli.

PubMed

Lee, Hoonsoo; Kim, Moon S; Qin, Jianwei; Park, Eunsoo; Song, Yu-Rim; Oh, Chang-Sik; Cho, Byoung-Kwan

2017-09-23

The bacterial infection of seeds is one of the most important quality factors affecting yield. Conventional detection methods for bacteria-infected seeds, such as biological, serological, and molecular tests, are not feasible since they require expensive equipment, and furthermore, the testing processes are also time-consuming. In this study, we use the Raman hyperspectral imaging technique to distinguish bacteria-infected seeds from healthy seeds as a rapid, accurate, and non-destructive detection tool. We utilize Raman hyperspectral imaging data in the spectral range of 400-1800 cm -1 to determine the optimal band-ratio for the discrimination of watermelon seeds infected by the bacteria Acidovorax citrulli using ANOVA. Two bands at 1076.8 cm -1 and 437 cm -1 are selected as the optimal Raman peaks for the detection of bacteria-infected seeds. The results demonstrate that the Raman hyperspectral imaging technique has a good potential for the detection of bacteria-infected watermelon seeds and that it could form a suitable alternative to conventional methods.

Recent Trends in Rapid Environmental Monitoring of Pathogens and Toxicants: Potential of Nanoparticle-Based Biosensor and Applications

PubMed Central

Koedrith, Preeyaporn; Thasiphu, Thalisa; Weon, Jong-Il; Boonprasert, Rattana; Tuitemwong, Kooranee; Tuitemwong, Pravate

2015-01-01

Of global concern, environmental pollution adversely affects human health and socioeconomic development. The presence of environmental contaminants, especially bacterial, viral, and parasitic pathogens and their toxins as well as chemical substances, poses serious public health concerns. Nanoparticle-based biosensors are considered as potential tools for rapid, specific, and highly sensitive detection of the analyte of interest (both biotic and abiotic contaminants). In particular, there are several limitations of conventional detection methods for water-borne pathogens due to low concentrations and interference with various enzymatic inhibitors in the environmental samples. The increase of cells to detection levels requires long incubation time. This review describes current state of biosensor nanotechnology, the advantage over conventional detection methods, and the challenges due to testing of environmental samples. The major approach is to use nanoparticles as signal reporter to increase output rather than spending time to increase cell concentrations. Trends in future development of novel detection devices and their advantages over other environmental monitoring methodologies are also discussed. PMID:25884032

Raman Hyperspectral Imaging for Detection of Watermelon Seeds Infected with Acidovorax citrulli

PubMed Central

Lee, Hoonsoo; Kim, Moon S.; Qin, Jianwei; Park, Eunsoo; Song, Yu-Rim; Oh, Chang-Sik

2017-01-01

The bacterial infection of seeds is one of the most important quality factors affecting yield. Conventional detection methods for bacteria-infected seeds, such as biological, serological, and molecular tests, are not feasible since they require expensive equipment, and furthermore, the testing processes are also time-consuming. In this study, we use the Raman hyperspectral imaging technique to distinguish bacteria-infected seeds from healthy seeds as a rapid, accurate, and non-destructive detection tool. We utilize Raman hyperspectral imaging data in the spectral range of 400–1800 cm−1 to determine the optimal band-ratio for the discrimination of watermelon seeds infected by the bacteria Acidovorax citrulli using ANOVA. Two bands at 1076.8 cm−1 and 437 cm−1 are selected as the optimal Raman peaks for the detection of bacteria-infected seeds. The results demonstrate that the Raman hyperspectral imaging technique has a good potential for the detection of bacteria-infected watermelon seeds and that it could form a suitable alternative to conventional methods. PMID:28946608

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

Identification and quantification of genetically modified Moonshade carnation lines using conventional and TaqMan real-time polymerase chain reaction methods.

PubMed

Li, Peng; Jia, Junwei; Bai, Lan; Pan, Aihu; Tang, Xueming

2013-07-01

Genetically modified carnation (Dianthus caryophyllus L.) Moonshade was approved for planting and commercialization in several countries from 2004. Developing methods for analyzing Moonshade is necessary for implementing genetically modified organism labeling regulations. In this study, the 5'-transgene integration sequence was isolated using thermal asymmetric interlaced (TAIL)-PCR. Based upon the 5'-transgene integration sequence, conventional and TaqMan real-time PCR assays were established. The relative limit of detection for the conventional PCR assay was 0.05 % for Moonshade using 100 ng total carnation genomic DNA, corresponding to approximately 79 copies of the carnation haploid genome, and the limits of detection and quantification of the TaqMan real-time PCR assay were estimated to be 51 and 254 copies of haploid carnation genomic DNA, respectively. These results are useful for identifying and quantifying Moonshade and its derivatives.

Early detection of mine fire in underground by using smell detectors

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

Ohga, Kotaro; Higuchi, Kiyoshi

1995-12-31

In our laboratory, a new detection system using smell detectors was developed to detect the spontaneous combustion of coal and the combustion of other materials used underground. Laboratory experiments were carried out on several kinds of coals, including South African coals, and machine oil, wood and rubber used in belt conveyers. The following results were obtained: (1) Spontaneous combustion of coal can be detected earlier by smell detectors than by conventional CO detection methods. (2) There were no differences in the results using different kinds of coal. (3) Combustion d other materials can also be detected earlier by this systemmore » than by conventional detectors for gas and smoke. (4) Use of this detection system enables one to discern the source of the combustion gases, whether it be coal, wood, oil or rubber.« less

Comparison of conventional culture method and fluorescent in situ hybridization technique for detection of Listeria spp. in ground beef, turkey, and chicken breast fillets in İzmir, Turkey.

PubMed

Baysal, Ayse Handan

2014-12-01

The occurrence of Listeria species in refrigerated fresh chicken breast fillet, turkey breast fillet, and ground beef was evaluated, comparing the conventional culture method and fluorescent in situ hybridization (FISH). FISH uses hybridization of a nucleic acid sequence target of a microorganism with a specific DNA probe labeled with a fluorochrome and imaging by a fluorescence microscope. First, Listeria was inoculated in chicken breast fillet, turkey breast fillet, or ground beef, and the applicability of the FISH method was evaluated. Second, Listeria was detected in fresh chicken breast fillet, turkey breast fillet, and ground beef by culture and FISH methods. Listeria was isolated from 27 (37.4%) of 216 samples by the standard culture method, whereas FISH detected 25 (24.7%) preenriched samples. Of these isolates, 17 (63%) were L. innocua, 6 (22%) L. welshimeri, and 4 (14.8%) L. seeligeri. Overall, the prevalences of Listeria spp. found with the conventional culture method in chicken breast fillet, turkey breast fillet, and ground beef were 9.7, 6.9, and 20.8%, whereas with the FISH technique these values were 11.1, 6.9, and 16.7%, respectively. The molecular FISH technique appears to be a cheap, sensitive, and time-efficient procedure that could be used for routine detection of Listeria spp. in meat. This study showed that retail raw meats are potentially contaminated with Listeria spp. and are, thus, vehicles for transmitting diseases caused by foodborne pathogens, underlining the need for increased precautions, such as implementation of hazard analysis and critical control points and consumer food safety education.

[Evaluation of a Computer-Aided Microscope System and Its Anti-Nuclear Antibody Test Kit for Indirect Immunofluorescence Assay].

PubMed

Hayashi, Nobuhide; Saegusa, Jun; Uto, Kenichi; Oyabu, Chinami; Saito, Toshiharu; Sato, Itsuko; Kawano, Seiji; Kumagai, Shunichi

2016-02-01

Antinuclear antibody (ANA) testing is indispensable for diagnosing and understanding clinical conditions of autoimmune diseases. The indirect immunofluorescence assay (IFA) is the gold standard for ANA screening, and it can detect more than 100 different antibodies, such as anti-PCNA as well as anti-cytoplasmic antibodies. However, complicated procedures of conventional IFA and visual interpretation require highly skilled laboratory staff. This study evaluates the capability, characteristics, and applicability of the recently developed ANA detection system (EUROPattern Cosmic IFA System, EPA) using HEp20-10 cells and the automated pattern recognition microscope. Findings using EPA and conventional methods were compared in 282 sera obtained from connective tissue disease patients and 250 sera from healthy individuals. The concordance of the positivity rate, antibody titer (within +/- 1 tube difference), and the accurate recognition rate of ANA patterns between the automated EPA method and the microscopic judgement of the EPA image by eye was 98.9, 97.4, and 55.3%, respectively. The EPA method showed concordance of the positivity rate as high as 93.3% and concordance of the antibody titer as high as 94.0% (within +/- 1 titer) compared with the conventional method. Regarding the four typical patterns of ANA (homogeneous, speckled, nucleolar, and centromere), large differences between the EPA and conventional methods were not observed, and the rate of concordance between the final EPA result and the conventional method was from 94.1 to 100%. The positivity rate of ANA using the EPA and conventional methods showed marked agreement among the six connective tissue diseases (SLE, MCTD, SSc, PM/DM, and SS) and healthy individuals. Although the EPA system is not considered a complete system and laboratory staff should verify the results, it is a useful system for routine ANA analysis because it contributes to ANA standardization and an efficient workflow.

QTL mapping for downy mildew resistance in cucumber via bulked segregant analysis using next-generation sequencing and conventional methods.

PubMed

Win, Khin Thanda; Vegas, Juan; Zhang, Chunying; Song, Kihwan; Lee, Sanghyeob

2017-01-01

QTL mapping using NGS-assisted BSA was successfully applied to an F 2 population for downy mildew resistance in cucumber. QTLs detected by NGS-assisted BSA were confirmed by conventional QTL analysis. Downy mildew (DM), caused by Pseudoperonospora cubensis, is one of the most destructive foliar diseases in cucumber. QTL mapping is a fundamental approach for understanding the genetic inheritance of DM resistance in cucumber. Recently, many studies have reported that a combination of bulked segregant analysis (BSA) and next-generation sequencing (NGS) can be a rapid and cost-effective way of mapping QTLs. In this study, we applied NGS-assisted BSA to QTL mapping of DM resistance in cucumber and confirmed the results by conventional QTL analysis. By sequencing two DNA pools each consisting of ten individuals showing high resistance and susceptibility to DM from a F 2 population, we identified single nucleotide polymorphisms (SNPs) between the two pools. We employed a statistical method for QTL mapping based on these SNPs. Five QTLs, dm2.2, dm4.1, dm5.1, dm5.2, and dm6.1, were detected and dm2.2 showed the largest effect on DM resistance. Conventional QTL analysis using the F 2 confirmed dm2.2 (R 2  = 10.8-24 %) and dm5.2 (R 2  = 14-27.2 %) as major QTLs and dm4.1 (R 2  = 8 %) as two minor QTLs, but could not detect dm5.1 and dm6.1. A new QTL on chromosome 2, dm2.1 (R 2  = 28.2 %) was detected by the conventional QTL method using an F 3 population. This study demonstrated the effectiveness of NGS-assisted BSA for mapping QTLs conferring DM resistance in cucumber and revealed the unique genetic inheritance of DM resistance in this population through two distinct major QTLs on chromosome 2 that mainly harbor DM resistance.

Enhancing the Detection of Giardia duodenalis Cysts in Foods by Inertial Microfluidic Separation

PubMed Central

Ganz, Kyle R.; Clime, Liviu; Farber, Jeffrey M.; Corneau, Nathalie

2015-01-01

The sensitivity and specificity of current Giardia cyst detection methods for foods are largely determined by the effectiveness of the elution, separation, and concentration methods used. The aim of these methods is to produce a final suspension with an adequate concentration of Giardia cysts for detection and a low concentration of interfering food debris. In the present study, a microfluidic device, which makes use of inertial separation, was designed and fabricated for the separation of Giardia cysts. A cyclical pumping platform and protocol was developed to concentrate 10-ml suspensions down to less than 1 ml. Tests involving Giardia duodenalis cysts and 1.90-μm microbeads in pure suspensions demonstrated the specificity of the microfluidic chip for cysts over smaller nonspecific particles. As the suspension cycled through the chip, a large number of beads were removed (70%) and the majority of the cysts were concentrated (82%). Subsequently, the microfluidic inertial separation chip was integrated into a method for the detection of G. duodenalis cysts from lettuce samples. The method greatly reduced the concentration of background debris in the final suspensions (10-fold reduction) in comparison to that obtained by a conventional method. The method also recovered an average of 68.4% of cysts from 25-g lettuce samples and had a limit of detection (LOD) of 38 cysts. While the recovery of cysts by inertial separation was slightly lower, and the LOD slightly higher, than with the conventional method, the sample analysis time was greatly reduced, as there were far fewer background food particles interfering with the detection of cysts by immunofluorescence microscopy. PMID:25841016

Detecting proteins in highly autofluorescent cells using quantum dot antibody conjugates.

PubMed

Orcutt, Karen M; Ren, Shanshan; Gundersen, Kjell

2009-01-01

We have applied quantum dot (Qdot) antibody conjugates as a biomolecular probe for cellular proteins important in biogeochemical cycling in the sea. Conventional immunological methods have been hampered by the strong autofluorescence found in cyanobacteria cells. Qdot conjugates provide an ideal alternative for studies that require long-term imaging of cells such as detection of low abundance cellular antigens by fluorescence microscopy. The advantage of Qdot labeled probes over conventional immunological methods is the photostability of the probe. Phycoerythrin bleaches in cyanobacterial cells under prolonged UV or blue light excitation, which means that the semiconducting nanocrystal probe, the Qdot, can yield a strong fluorescent signal without interference from cellular pigments.

Application of nanomaterials in the bioanalytical detection of disease-related genes.

PubMed

Zhu, Xiaoqian; Li, Jiao; He, Hanping; Huang, Min; Zhang, Xiuhua; Wang, Shengfu

2015-12-15

In the diagnosis of genetic diseases and disorders, nanomaterials-based gene detection systems have significant advantages over conventional diagnostic systems in terms of simplicity, sensitivity, specificity, and portability. In this review, we describe the application of nanomaterials for disease-related genes detection in different methods excluding PCR-related method, such as colorimetry, fluorescence-based methods, electrochemistry, microarray methods, surface-enhanced Raman spectroscopy (SERS), quartz crystal microbalance (QCM) methods, and dynamic light scattering (DLS). The most commonly used nanomaterials are gold, silver, carbon and semiconducting nanoparticles. Various nanomaterials-based gene detection methods are introduced, their respective advantages are discussed, and selected examples are provided to illustrate the properties of these nanomaterials and their emerging applications for the detection of specific nucleic acid sequences. Copyright © 2015. Published by Elsevier B.V.

Detection of Salmonella sp in chicken cuts using immunomagnetic separation

PubMed Central

de Cássia dos Santos da Conceição, Rita; Moreira, Ângela Nunes; Ramos, Roberta Juliano; Goularte, Fabiana Lemos; Carvalhal, José Beiro; Aleixo, José Antonio Guimarães

2008-01-01

The immunomagnetic separation (IMS) is a technique that has been used to increase sensitivity and specificity and to decrease the time required for detection of Salmonella in foods through different methodologies. In this work we report on the development of a method for detection of Salmonella in chicken cuts using in house antibody-sensitized microspheres associated to conventional plating in selective agar (IMS-plating). First, protein A-coated microspheres were sensitized with polyclonal antibodies against lipopolysacharide and flagella from salmonellae and used to standardize a procedure for capturing Salmonella Enteritidis from pure cultures and detection in selective agar. Subsequently, samples of chicken meat experimentally contaminated with S. Enteritidis were analyzed immediately after contamination and after 24h of refrigeration using three enrichment protocols. The detection limit of the IMS-plating procedure after standardization with pure culture was about 2x10 CFU/mL. The protocol using non-selective enrichment for 6-8h, selective enrichment for 16-18h and a post-enrichment for 4h gave the best results of S. Enteritidis detection by IMS-plating in experimentally contaminated meat. IMS-plating using this protocol was compared to the standard culture method for salmonellae detection in naturally contaminated chicken cuts and yielded 100% sensitivity and 94% specificity. The method developed using in house prepared magnetic microespheres for IMS and plating in selective agar was able to diminish by at least one day the time required for detection of Salmonella in chicken products by the conventional culture method. PMID:24031199

Use of vectors in sequence analysis.

PubMed

Ishikawa, T; Yamamoto, K; Yoshikura, H

1987-10-01

Applications of the vector diagram, a new type of representation of protein structure, in homology search of various proteins including oncogene products are presented. The method takes account of various kinds of information concerning the properties of amino acids, such as Chou and Fasman's probability data. The method can detect conformational similarities of proteins which may not be detected by the conventional programs.

Task-based statistical image reconstruction for high-quality cone-beam CT

NASA Astrophysics Data System (ADS)

Dang, Hao; Webster Stayman, J.; Xu, Jennifer; Zbijewski, Wojciech; Sisniega, Alejandro; Mow, Michael; Wang, Xiaohui; Foos, David H.; Aygun, Nafi; Koliatsos, Vassilis E.; Siewerdsen, Jeffrey H.

2017-11-01

Task-based analysis of medical imaging performance underlies many ongoing efforts in the development of new imaging systems. In statistical image reconstruction, regularization is often formulated in terms to encourage smoothness and/or sharpness (e.g. a linear, quadratic, or Huber penalty) but without explicit formulation of the task. We propose an alternative regularization approach in which a spatially varying penalty is determined that maximizes task-based imaging performance at every location in a 3D image. We apply the method to model-based image reconstruction (MBIR—viz., penalized weighted least-squares, PWLS) in cone-beam CT (CBCT) of the head, focusing on the task of detecting a small, low-contrast intracranial hemorrhage (ICH), and we test the performance of the algorithm in the context of a recently developed CBCT prototype for point-of-care imaging of brain injury. Theoretical predictions of local spatial resolution and noise are computed via an optimization by which regularization (specifically, the quadratic penalty strength) is allowed to vary throughout the image to maximize local task-based detectability index ({{d}\\prime} ). Simulation studies and test-bench experiments were performed using an anthropomorphic head phantom. Three PWLS implementations were tested: conventional (constant) penalty; a certainty-based penalty derived to enforce constant point-spread function, PSF; and the task-based penalty derived to maximize local detectability at each location. Conventional (constant) regularization exhibited a fairly strong degree of spatial variation in {{d}\\prime} , and the certainty-based method achieved uniform PSF, but each exhibited a reduction in detectability compared to the task-based method, which improved detectability up to ~15%. The improvement was strongest in areas of high attenuation (skull base), where the conventional and certainty-based methods tended to over-smooth the data. The task-driven reconstruction method presents a promising regularization method in MBIR by explicitly incorporating task-based imaging performance as the objective. The results demonstrate improved ICH conspicuity and support the development of high-quality CBCT systems.

Nucleic acid amplification tests (NAATs) for gonorrhoea diagnosis in women: experience of a tertiary care hospital in north India.

PubMed

Sood, Seema; Verma, Rachna; Mir, Shazia Shaheen; Agarwal, Madhav; Singh, Neeta; Kar, Hemanta Kumar; Sharma, Vinod Kumar

2014-11-01

Gonorrhoea is among the most frequent of the estimated bacterial sexually transmitted infections (STIs) and has significant health implications in women. The use of nucleic acid amplification tests (NAATs) has been shown to provide enhanced diagnosis of gonorrhoea in female patients. However, it is recommended that an on-going assessment of the test assays should be performed to check for any probable sequence variation occurring in the targeted region. In this study, an in-house PCR targeting opa-gene of Neisseria gonorrhoeae was used in conjunction with 16S ribosomal PCR to determine the presence of gonorrhoea in female patients attending the tertiary care hospitals. Endocervical samples collected from 250 female patients with complaints of vaginal or cervical discharge or pain in lower abdomen were tested using opa and 16S ribosomal assay. The samples were also processed by conventional methods. Of the 250 female patients included in the study, only one was positive by conventional methods (microscopy and culture) whereas 17 patients were found to be positive based on PCR results. The clinical sensitivity of conventional methods for the detection of N. gonorrhoeae in female patients was low. The gonococcal detection rates increased when molecular method was used giving 16 additional positives. Studies should be done to find out other gene targets that may be used in the screening assays to detect the presence of gonorrhoea.

Influence of Detection Methods in Characterizing Escherichia coli O157:H7 in Raw Goat Meat Using Conventional and Molecular Methods.

PubMed

Tabashsum, Zajeba; Nazneen, Mafruha; Ahsan, C R; Bari, M L; Yasmin, M

2016-01-01

　Presence of Escherichia coli O157:H7 on fresh goat meat samples (n= 40) of Dhaka city was analyzed using conventional and molecular methods. A total of 86 presumptive E. coli O157:H7 colonies were isolated from 60% of the samples using selective agar plating method. After conventional biochemical assay followed by API 20E assay, only 11 isolates were found to be E. coli O157:H7. Further serological test identified only four isolates that has strong agglutination reaction against anti-H7 sensitized latex. The biochemically and serologically confirmed isolates were then screened for major virulence factors include eaeA, rfbE, fliC, stx1 and stx2 genes by PCR. PCR analysis of positive isolates showed, 10 isolates were eaeA and rfbE genes positive but fliC gene was only in six, indicating that these isolates were H7 positive with flagellum antigens which might not expressed or detected in serotyping tests. Multiplex PCR against eaeA, stx1 and stx2 genes of the isolates showed similar results as when done individually. These results revealed that only 7% of the primary presumptive E. coli O157:H7 was found to be stx producing E. coli O157:H7 and thus greatly influenced the detection of the pathogen in meat samples.

Real-time PCR assay is superior to other methods for the detection of mycoplasma contamination in the cell lines of the National Cell Bank of Iran.

PubMed

Molla Kazemiha, Vahid; Bonakdar, Shahin; Amanzadeh, Amir; Azari, Shahram; Memarnejadian, Arash; Shahbazi, Shirin; Shokrgozar, Mohammad Ali; Mahdian, Reza

2016-08-01

Mycoplasmas are the most important contaminants of cell cultures throughout the world. They are considered as a major problem in biological studies and biopharmaceutical economic issues. In this study, our aim was to find the best standard technique as a rapid method with high sensitivity, specificity and accuracy for the detection of mycoplasma contamination in the cell lines of the National Cell Bank of Iran. Thirty cell lines suspected to mycoplasma contamination were evaluated by five different techniques including microbial culture, indirect DNA DAPI staining, enzymatic mycoalert(®) assay, conventional PCR and real-time PCR. Five mycoplasma-contaminated cell lines were assigned as positive controls and five mycoplasma-free cell lines as negative controls. The enzymatic method was performed using the mycoalert(®) mycoplasma detection kit. Real-time PCR technique was conducted by PromoKine diagnostic kits. In the conventional PCR method, mycoplasma genus-specific primers were designed to analyze the sequences based on a fixed and common region on 16S ribosomal RNA with PCR product size of 425 bp. Mycoplasma contamination was observed in 60, 56.66, 53.33, 46.66 and 33.33 % of 30 different cell cultures by real-time PCR, PCR, enzymatic mycoalert(®), indirect DNA DAPI staining and microbial culture methods, respectively. The analysis of the results of the different methods showed that the real-time PCR assay was superior the other methods with the sensitivity, specificity, accuracy, predictive value of positive and negative results of 100 %. These values were 94.44, 100, 96.77, 100 and 92.85 % for the conventional PCR method, respectively. Therefore, this study showed that real-time PCR and PCR assays based on the common sequences in the 16S ribosomal RNA are reliable methods with high sensitivity, specificity and accuracy for detection of mycoplasma contamination in cell cultures and other biological products.

An intelligent detection method for high-field asymmetric waveform ion mobility spectrometry.

PubMed

Li, Yue; Yu, Jianwen; Ruan, Zhiming; Chen, Chilai; Chen, Ran; Wang, Han; Liu, Youjiang; Wang, Xiaozhi; Li, Shan

2018-04-01

In conventional high-field asymmetric waveform ion mobility spectrometry signal acquisition, multi-cycle detection is time consuming and limits somewhat the technique's scope for rapid field detection. In this study, a novel intelligent detection approach has been developed in which a threshold was set on the relative error of α parameters, which can eliminate unnecessary time spent on detection. In this method, two full-spectrum scans were made in advance to obtain the estimated compensation voltage at different dispersion voltages, resulting in a narrowing down of the whole scan area to just the peak area(s) of interest. This intelligent detection method can reduce the detection time to 5-10% of that of the original full-spectrum scan in a single cycle.

Interactive-predictive detection of handwritten text blocks

NASA Astrophysics Data System (ADS)

Ramos Terrades, O.; Serrano, N.; Gordó, A.; Valveny, E.; Juan, A.

2010-01-01

A method for text block detection is introduced for old handwritten documents. The proposed method takes advantage of sequential book structure, taking into account layout information from pages previously transcribed. This glance at the past is used to predict the position of text blocks in the current page with the help of conventional layout analysis methods. The method is integrated into the GIDOC prototype: a first attempt to provide integrated support for interactive-predictive page layout analysis, text line detection and handwritten text transcription. Results are given in a transcription task on a 764-page Spanish manuscript from 1891.

Detection of Salmonella invA gene in shrimp enrichment culture by polymerase chain reaction.

PubMed

Upadhyay, Bishnu Prasad; Utrarachkij, Fuangfa; Thongshoob, Jarinee; Mahakunkijcharoen, Yuvadee; Wongchinda, Niracha; Suthienkul, Orasa; Khusmith, Srisin

2010-03-01

Contamination of seafood with salmonellae is a major public health concern. Detection of Salmonella by standard culture methods is time consuming. In this study, an enrichment culture step prior to polymerase chain reaction (PCR) was applied to detect 284 bp fragment of Salmonella invA in comparison with the conventional culture method in 100 shrimp samples collected from four different shrimp farms and fresh food markets around Bangkok. Samples were pre-enriched in non-selective lactose broth (LB) and selective tetrathionate broth (TTB). PCR detection limit was 10 pg and 10(4) cfu/ml of viable salmonellae with 100% specificity. PCR assay detected 19 different Salmonella serovars belonging to 8 serogroups (B, C1, C2-C3, D1, E1, E4 and K) commonly found in clinical and environmental samples in Thailand. The detection rate of PCR following TTB enrichment (24%) was higher than conventional culture method (19%). PCR following TTB, but not in LB enrichment allowed salmonella detection with 84% sensitivity, 90% specificity and 89% accuracy. Shrimp samples collected from fresh food markets had higher levels of contaminated salmonellae than those from shrimp farms. The results indicated that incorporation of an enrichment step prior to PCR has the potential to be applied for detection of naturally contaminated salmonellae in food, environment and clinical samples.

Development of a panel of seven duplex real-time PCR assays for detecting 13 streptococcal superantigens.

PubMed

Yang, Peng; Peng, Xiaomin; Cui, Shujuan; Shao, Junbin; Zhu, Xuping; Zhang, Daitao; Liang, Huijie; Wang, Quanyi

2013-07-30

Streptococcal superantigens (SAgs) are the major virulence factors of infection in humans for group A Streptococcus (GAS) bacteria. A panel consisting of seven duplex real-time PCR assays was developed to simultaneously detect 13 streptococcal SAgs and one internal control which may be important in the control of GAS-mediated diseases. Primer and probe sequences were selected based on the highly conserved region from an alignment of nucleotide sequences of the 13 streptococcal SAgs. The reaction conditions of the duplex real-time PCR were optimized and the specificity of the duplex assays was evaluated using SAg positive strains. The limit of detection of the duplex assays was determined by using 10-fold serial dilutions of the DNA of 13 streptococcal SAgs and compared to a conventional polymerase chain reaction (PCR) method for evaluating the duplex assays sensitivity. Using the duplex assays, we were able to differentiate between 13 SAgs from Streptococcus strains and other non-Streptococcus bacteria without cross-reaction. On the other hand, the limit of detection of the duplex assays was at least one or two log dilutions lower than that of the conventional PCR. The panel was highly specific (100%) and the limit of detection of these duplex groups was at least ten times lower than that obtained by using a conventional PCR method.

Detection of fresh bruises in apples by structured-illumination reflectance imaging

NASA Astrophysics Data System (ADS)

Lu, Yuzhen; Li, Richard; Lu, Renfu

2016-05-01

Detection of fresh bruises in apples remains a challenging task due to the absence of visual symptoms and significant chemical alterations of fruit tissues during the initial stage after the fruit have been bruised. This paper reports on a new structured-illumination reflectance imaging (SIRI) technique for enhanced detection of fresh bruises in apples. Using a digital light projector engine, sinusoidally-modulated illumination at the spatial frequencies of 50, 100, 150 and 200 cycles/m was generated. A digital camera was then used to capture the reflectance images from `Gala' and `Jonagold' apples, immediately after they had been subjected to two levels of bruising by impact tests. A conventional three-phase demodulation (TPD) scheme was applied to the acquired images for obtaining the planar (direct component or DC) and amplitude (alternating component or AC) images. Bruises were identified in the amplitude images with varying image contrasts, depending on spatial frequency. The bruise visibility was further enhanced through post-processing of the amplitude images. Furthermore, three spiral phase transform (SPT)-based demodulation methods, using single and two images and two phase-shifted images, were proposed for obtaining AC images. Results showed that the demodulation methods greatly enhanced the contrast and spatial resolution of the AC images, making it feasible to detect the fresh bruises that, otherwise, could not be achieved by conventional imaging technique with planar or uniform illumination. The effectiveness of image enhancement, however, varied with spatial frequency. Both 2-image and 2-phase SPT methods achieved the performance similar to that by conventional TPD. SIRI technique has demonstrated the capability of detecting fresh bruises in apples, and it has the potential as a new imaging modality for enhancing food quality and safety detection.

A multiplex PCR method for the identification of commercially important salmon and trout species (Oncorhynchus and Salmo) in North America.

PubMed

Rasmussen Hellberg, Rosalee S; Morrissey, Michael T; Hanner, Robert H

2010-09-01

The purpose of this study was to develop a species-specific multiplex polymerase chain reaction (PCR) method that allows for the detection of salmon species substitution on the commercial market. Species-specific primers and TaqMan® probes were developed based on a comprehensive collection of mitochondrial 5' cytochrome c oxidase subunit I (COI) deoxyribonucleic acid (DNA) "barcode" sequences. Primers and probes were combined into multiplex assays and tested for specificity against 112 reference samples representing 25 species. Sensitivity and linearity tests were conducted using 10-fold serial dilutions of target DNA (single-species samples) and DNA admixtures containing the target species at levels of 10%, 1.0%, and 0.1% mixed with a secondary species. The specificity tests showed positive signals for the target DNA in both real-time and conventional PCR systems. Nonspecific amplification in both systems was minimal; however, false positives were detected at low levels (1.2% to 8.3%) in conventional PCR. Detection levels were similar for admixtures and single-species samples based on a 30 PCR cycle cut-off, with limits of 0.25 to 2.5 ng (1% to 10%) in conventional PCR and 0.05 to 5.0 ng (0.1% to 10%) in real-time PCR. A small-scale test with food samples showed promising results, with species identification possible even in heavily processed food items. Overall, this study presents a rapid, specific, and sensitive method for salmon species identification that can be applied to mixed-species and heavily processed samples in either conventional or real-time PCR formats. This study provides a newly developed method for salmon and trout species identification that will assist both industry and regulatory agencies in the detection and prevention of species substitution. This multiplex PCR method allows for rapid, high-throughput species identification even in heavily processed and mixed-species samples. An inter-laboratory study is currently being carried out to assess the ability of this method to identify species in a variety of commercial salmon and trout products.

A multiplex PCR assay for the rapid and sensitive detection of methicillin-resistant Staphylococcus aureus and simultaneous discrimination of Staphylococcus aureus from coagulase-negative staphylococci.

PubMed

Xu, Benjin; Liu, Ling; Liu, Li; Li, Xinping; Li, Xiaofang; Wang, Xin

2012-11-01

Methicillin-resistant Staphylococcus aureus (MRSA) is a global health concern, which had been detected in food and food production animals. Conventional testing for detection of MRSA takes 3 to 5 d to yield complete information of the organism and its antibiotic sensitivity pattern. So, a rapid method is needed to diagnose and treat the MRSA infections. The present study focused on the development of a multiplex PCR assay for the rapid and sensitive detection of MRSA. The assay simultaneously detected 4 genes, namely, 16S rRNA of the Staphylococcus genus, femA of S. aureus, mecA that encodes methicillin resistance, and one internal control. It was rapid and yielded results within 4 h. The analytical sensitivity and specificity of the multiplex PCR assay was evaluated by comparing it with the conventional method. The analytical sensitivity of the multiplex PCR assay at the DNA level was 10 ng DNA. The analytical specificity was evaluated with 10 reference staphylococci strains and was 100%. The diagnostic evaluation of MRSA was carried out using 360 foodborne staphylococci isolates, and showed 99.1% of specificity, 96.4% of sensitivity, 97.5% of positive predictive value, and 97.3% of negative predictive value compared to the conventional method. The inclusion of an internal control in the multiplex PCR assay is important to exclude false-negative cases. This test can be used as an effective diagnostic and surveillance tool to investigate the spread and emergence of MRSA. © 2012 Institute of Food Technologists®

Detection of premature ventricular contractions on a ventricular electrocardiogram for patients with left ventricular assist devices.

PubMed

Park, Sung Min; Lee, Jin Hong; Choi, Seong Wook

2014-12-01

The ventricular electrocardiogram (v-ECG) was developed for long-term monitoring of heartbeats in patients with a left ventricular assist device (LVAD) and does not normally have the functionality necessary to detect additional heart irregularities that can progress to critical arrhythmias. Although the v-ECG has the benefits of physiological optimization and counterpulsation control, when abnormal heartbeats occur, the v-ECG does not show the distinct abnormal waveform that enables easy detection of an abnormal heartbeat among normal heartbeats on the conventional ECG. In this study, the v-ECGs of normal and abnormal heartbeats are compared with each other with respect to peak-to-peak voltage, area, and maximal slopes, and a new method to detect abnormal heartbeats is suggested. In a series of animal experiments with three porcine models (Yorkshire pigs weighing 30-40 kg), a v-ECG and conventional ECG were taken simultaneously during LVAD perfusion. Clinical experts found 104 abnormal heartbeats from the saved conventional ECG data and confirmed that the other 3159 heartbeats were normal. Almost all of the abnormal heartbeats were premature ventricular contractions (PVCs), and there was short-term tachycardia for 3 s. A personal computer was used to automatically detect abnormal heartbeats with the v-ECG according to the new method, and its results were compared with the clinicians' results. The new method found abnormal heartbeats with 90% accuracy, and less than 15% of the total PVCs were missed. Copyright © 2014 International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

Rapid detection of the CYP2A6*12 hybrid allele by Pyrosequencing technology.

PubMed

Koontz, Deborah A; Huckins, Jacqueline J; Spencer, Antonina; Gallagher, Margaret L

2009-08-24

Identification of CYP2A6 alleles associated with reduced enzyme activity is important in the study of inter-individual differences in drug metabolism. CYP2A6*12 is a hybrid allele that results from unequal crossover between CYP2A6 and CYP2A7 genes. The 5' regulatory region and exons 1-2 are derived from CYP2A7, and exons 3-9 are derived from CYP2A6. Conventional methods for detection of CYP2A6*12 consist of two-step PCR protocols that are laborious and unsuitable for high-throughput genotyping. We developed a rapid and accurate method to detect the CYP2A6*12 allele by Pyrosequencing technology. A single set of PCR primers was designed to specifically amplify both the CYP2A6*1 wild-type allele and the CYP2A6*12 hybrid allele. An internal Pyrosequencing primer was used to generate allele-specific sequence information, which detected homozygous wild-type, heterozygous hybrid, and homozygous hybrid alleles. We first validated the assay on 104 DNA samples that were also genotyped by conventional two-step PCR and by cycle sequencing. CYP2A6*12 allele frequencies were then determined using the Pyrosequencing assay on 181 multi-ethnic DNA samples from subjects of African American, European Caucasian, Pacific Rim, and Hispanic descent. Finally, we streamlined the Pyrosequencing assay by integrating liquid handling robotics into the workflow. Pyrosequencing results demonstrated 100% concordance with conventional two-step PCR and cycle sequencing methods. Allele frequency data showed slightly higher prevalence of the CYP2A6*12 allele in European Caucasians and Hispanics. This Pyrosequencing assay proved to be a simple, rapid, and accurate alternative to conventional methods, which can be easily adapted to the needs of higher-throughput studies.

Earlier Detection of Tumor Treatment Response Using Magnetic Resonance Diffusion Imaging with Oscillating Gradients

PubMed Central

Colvin, Daniel C.; Loveless, Mary E.; Does, Mark D.; Yue, Zou; Yankeelov, Thomas E.; Gore, John C.

2011-01-01

An improved method for detecting early changes in tumors in response to treatment, based on a modification of diffusion-weighted magnetic resonance imaging, has been demonstrated in an animal model. Early detection of therapeutic response in tumors is important both clinically and in pre-clinical assessments of novel treatments. Non-invasive imaging methods that can detect and assess tumor response early in the course of treatment, and before frank changes in tumor morphology are evident, are of considerable interest as potential biomarkers of treatment efficacy. Diffusion-weighted magnetic resonance imaging is sensitive to changes in water diffusion rates in tissues that result from structural variations in the local cellular environment, but conventional methods mainly reflect changes in tissue cellularity and do not convey information specific to micro-structural variations at sub-cellular scales. We implemented a modified imaging technique using oscillating gradients of the magnetic field for evaluating water diffusion rates over very short spatial scales that are more specific for detecting changes in intracellular structure that may precede changes in cellularity. Results from a study of orthotopic 9L gliomas in rat brains indicate that this method can detect changes as early as 24 hours following treatment with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), when conventional approaches do not find significant effects. These studies suggest that diffusion imaging using oscillating gradients may be used to obtain an earlier indication of treatment efficacy than previous magnetic resonance imaging methods. PMID:21190804

Direct Detection of Biotinylated Proteins by Mass Spectrometry

PubMed Central

2015-01-01

Mass spectrometric strategies to identify protein subpopulations involved in specific biological functions rely on covalently tagging biotin to proteins using various chemical modification methods. The biotin tag is primarily used for enrichment of the targeted subpopulation for subsequent mass spectrometry (MS) analysis. A limitation of these strategies is that MS analysis does not easily discriminate unlabeled contaminants from the labeled protein subpopulation under study. To solve this problem, we developed a flexible method that only relies on direct MS detection of biotin-tagged proteins called “Direct Detection of Biotin-containing Tags” (DiDBiT). Compared with conventional targeted proteomic strategies, DiDBiT improves direct detection of biotinylated proteins ∼200 fold. We show that DiDBiT is applicable to several protein labeling protocols in cell culture and in vivo using cell permeable NHS-biotin and incorporation of the noncanonical amino acid, azidohomoalanine (AHA), into newly synthesized proteins, followed by click chemistry tagging with biotin. We demonstrate that DiDBiT improves the direct detection of biotin-tagged newly synthesized peptides more than 20-fold compared to conventional methods. With the increased sensitivity afforded by DiDBiT, we demonstrate the MS detection of newly synthesized proteins labeled in vivo in the rodent nervous system with unprecedented temporal resolution as short as 3 h. PMID:25117199

SQUID-detected FMR: Resonance in single crystalline and polycrystalline yttrium iron garnet

NASA Astrophysics Data System (ADS)

O'Reilly, J. M.; Stamenov, P.

2018-04-01

Here two new techniques for the detection of broadband (100 MHz-20 GHz) ferromagnetic resonance (FMR)/ferrimagnetic resonance in single and poly-crystalline materials, which rely on SQUID-based gradiometry detection of small changes in the magnetisation, are developed. In the first method, small changes in the along-the-applied-field projection of the coupled magnetic moment (Δmz) are detected as the material is driven into resonance. Absolute measurement of the longitudinal component of the magnetisation and the resonance induced lowering of this moment makes estimation of the precession cone angle accessible, which is typically difficult to extract using conventional cavity or stripline based detection methods. The second method invokes the change in Δmz with the resonance-induced thermal heating (d/mz d T ). Magnetisation dynamics in bulk Y3Fe5O12 are observed over a broad range of experimental temperatures (4 K-400 K) and fields (10-500 mT). The inhomogeneous microwave excitation allows for the observation of higher magnetostatic modes and the convenient tracking of very broad resonances. The two SQUID-detection techniques when combined with conventional broadband vector network analyser-FMR, low-frequency magnetic susceptibility, and DC magnetometry, all easily realised, essentially concurrently, using the same module, greatly expand the amount of static and dynamic information accessible.

Developing a new case based computer-aided detection scheme and an adaptive cueing method to improve performance in detecting mammographic lesions

PubMed Central

Tan, Maxine; Aghaei, Faranak; Wang, Yunzhi; Zheng, Bin

2017-01-01

The purpose of this study is to evaluate a new method to improve performance of computer-aided detection (CAD) schemes of screening mammograms with two approaches. In the first approach, we developed a new case based CAD scheme using a set of optimally selected global mammographic density, texture, spiculation, and structural similarity features computed from all four full-field digital mammography (FFDM) images of the craniocaudal (CC) and mediolateral oblique (MLO) views by using a modified fast and accurate sequential floating forward selection feature selection algorithm. Selected features were then applied to a “scoring fusion” artificial neural network (ANN) classification scheme to produce a final case based risk score. In the second approach, we combined the case based risk score with the conventional lesion based scores of a conventional lesion based CAD scheme using a new adaptive cueing method that is integrated with the case based risk scores. We evaluated our methods using a ten-fold cross-validation scheme on 924 cases (476 cancer and 448 recalled or negative), whereby each case had all four images from the CC and MLO views. The area under the receiver operating characteristic curve was AUC = 0.793±0.015 and the odds ratio monotonically increased from 1 to 37.21 as CAD-generated case based detection scores increased. Using the new adaptive cueing method, the region based and case based sensitivities of the conventional CAD scheme at a false positive rate of 0.71 per image increased by 2.4% and 0.8%, respectively. The study demonstrated that supplementary information can be derived by computing global mammographic density image features to improve CAD-cueing performance on the suspicious mammographic lesions. PMID:27997380

Intelligent Method for Diagnosing Structural Faults of Rotating Machinery Using Ant Colony Optimization

PubMed Central

Li, Ke; Chen, Peng

2011-01-01

Structural faults, such as unbalance, misalignment and looseness, etc., often occur in the shafts of rotating machinery. These faults may cause serious machine accidents and lead to great production losses. This paper proposes an intelligent method for diagnosing structural faults of rotating machinery using ant colony optimization (ACO) and relative ratio symptom parameters (RRSPs) in order to detect faults and distinguish fault types at an early stage. New symptom parameters called “relative ratio symptom parameters” are defined for reflecting the features of vibration signals measured in each state. Synthetic detection index (SDI) using statistical theory has also been defined to evaluate the applicability of the RRSPs. The SDI can be used to indicate the fitness of a RRSP for ACO. Lastly, this paper also compares the proposed method with the conventional neural networks (NN) method. Practical examples of fault diagnosis for a centrifugal fan are provided to verify the effectiveness of the proposed method. The verification results show that the structural faults often occurring in the centrifugal fan, such as unbalance, misalignment and looseness states are effectively identified by the proposed method, while these faults are difficult to detect using conventional neural networks. PMID:22163833

Intelligent method for diagnosing structural faults of rotating machinery using ant colony optimization.

PubMed

Li, Ke; Chen, Peng

2011-01-01

Structural faults, such as unbalance, misalignment and looseness, etc., often occur in the shafts of rotating machinery. These faults may cause serious machine accidents and lead to great production losses. This paper proposes an intelligent method for diagnosing structural faults of rotating machinery using ant colony optimization (ACO) and relative ratio symptom parameters (RRSPs) in order to detect faults and distinguish fault types at an early stage. New symptom parameters called "relative ratio symptom parameters" are defined for reflecting the features of vibration signals measured in each state. Synthetic detection index (SDI) using statistical theory has also been defined to evaluate the applicability of the RRSPs. The SDI can be used to indicate the fitness of a RRSP for ACO. Lastly, this paper also compares the proposed method with the conventional neural networks (NN) method. Practical examples of fault diagnosis for a centrifugal fan are provided to verify the effectiveness of the proposed method. The verification results show that the structural faults often occurring in the centrifugal fan, such as unbalance, misalignment and looseness states are effectively identified by the proposed method, while these faults are difficult to detect using conventional neural networks.

Development of a micromanipulation method for single cell isolation of prokaryotes and its application in food safety.

PubMed

Hohnadel, Marisa; Maumy, Myriam; Chollet, Renaud

2018-01-01

For nearly a century, conventional microbiological methods have been standard practice for detecting and identifying pathogens in food. Nevertheless, the microbiological safety of food has improved and various rapid methods have been developed to overcome the limitations of conventional methods. Alternative methods are expected to detect low cell numbers, since the presence in food of even a single cell of a pathogenic organism may be infectious. With respect to low population levels, the performance of a detection method is assessed by producing serial dilutions of a pure bacterial suspension to inoculate representative food matrices with highly diluted bacterial cells (fewer than 10 CFU/ml). The accuracy of data obtained by multiple dilution techniques is not certain and does not exclude some colonies arising from clumps of cells. Micromanipulation techniques to capture and isolate single cells from environmental samples were introduced more than 40 years ago. The main limitation of the current micromanipulation technique is still the low recovery rate for the growth of a single cell in culture medium. In this study, we describe a new single cell isolation method and demonstrate that it can be used successfully to grow various types of microorganism from picked individual cells. Tests with Gram-positive and Gram-negative organisms, including cocci, rods, aerobes, anaerobes, yeasts and molds showed growth recovery rates from 60% to 100% after micromanipulation. We also highlight the use of our method to evaluate and challenge the detection limits of standard detection methods in food samples contaminated by a single cell of Salmonella enterica.

[Methicillin resistance detection in Staphylococcus aureus: comparison between conventional methods and MRSA-Screen latex agglutination technique].

PubMed

Soloaga, R; Corso, A; Gagetti, P; Faccone, D; Galas, M

2004-01-01

Methicillin-resistant Staphylococcus aureus (MRSA) is a significant pathogen that has emerged over the last four decades, causing both nosocomial and community-acquired infections. Rapid and accurate detection of methicillin resistance in S. aureus is important for the use of appropriate antimicrobial therapy and for the control of nosocomial spread of MRSA strains. We evaluated the efficiency of conventional methods for detection of methicillin resistance such as the disk diffusion, agar dilution, oxacillin agar screen test, and the latex agglutination test MRSA-Screen latex, in 100 isolates of S. aureus, 79 mecA positive and 21 mecA negative. The MRSA-Screen latex (Denka Seiken, Niigata, Japón), is a latex agglutination method that detects the presence of PLP-2a, product of mecA gene in S. aureus. The PCR of the mecA gene was used as the "gold standard" for the evaluation of the different methods tested. The percentages of sensitivity and specificity were as follows: disk difusión 97 and 100%, agar dilution 97 and 95%, oxacillin agar screen test 100 and 100%, and MRSA-Screen latex, 100 and 100 %. All methods presented high sensitivity and specificity, but MRSA-Screen latex had the advantage of giving a reliable result, equivalent to PCR, in only 15 minutes.

Detection of admittivity anomaly on high-contrast heterogeneous backgrounds using frequency difference EIT.

PubMed

Jang, J; Seo, J K

2015-06-01

This paper describes a multiple background subtraction method in frequency difference electrical impedance tomography (fdEIT) to detect an admittivity anomaly from a high-contrast background conductivity distribution. The proposed method expands the use of the conventional weighted frequency difference EIT method, which has been used limitedly to detect admittivity anomalies in a roughly homogeneous background. The proposed method can be viewed as multiple weighted difference imaging in fdEIT. Although the spatial resolutions of the output images by fdEIT are very low due to the inherent ill-posedness, numerical simulations and phantom experiments of the proposed method demonstrate its feasibility to detect anomalies. It has potential application in stroke detection in a head model, which is highly heterogeneous due to the skull.

Immunostimulatory oligonucleotide-induced metaphase cytogenetics detect chromosomal aberrations in 80% of CLL patients: A study of 132 CLL cases with correlation to FISH, IgVH status, and CD38 expression.

PubMed

Dicker, Frank; Schnittger, Susanne; Haferlach, Torsten; Kern, Wolfgang; Schoch, Claudia

2006-11-01

Compared with fluorescence in situ hybridization (FISH), conventional metaphase cytogenetics play only a minor prognostic role in chronic lymphocytic leukemia (CLL) so far, due to technical problems resulting from limited proliferation of CLL cells in vitro. Here, we present a simple method for in vitro stimulation of CLL cells that overcomes this limitation. In our unselected patient population, 125 of 132 cases could be successfully stimulated for metaphase generation by culture with the immunostimulatory CpG-oligonucleotide DSP30 plus interleukin 2. Of 125 cases, 101 showed chromosomal aberrations. The aberration rate is comparable to the rate detected by parallel interphase FISH. In 47 patients, conventional cytogenetics detected additional aberrations not detected by FISH analysis. A complex aberrant karyotype, defined as one having at least 3 aberrations, was detected in 30 of 125 patients, compared with only one such case as defined by FISH. Conventional cytogenetics frequently detected balanced and unbalanced translocations. A significant correlation of the poor-prognosis unmutated IgV(H) status with unbalanced translocations and of the likewise poor-prognosis CD38 expression to balanced translocations and complex aberrant karyotype was found. We demonstrate that FISH analysis underestimates the complexity of chromosomal aberrations in CLL. Therefore, conventional cytogenetics may define subgroups of patients with high risk of progression.

Application of polymerase chain reaction for detection of Vibrio parahaemolyticus associated with tropical seafoods and coastal environment.

PubMed

Dileep, V; Kumar, H S; Kumar, Y; Nishibuchi, M; Karunasagar, Indrani; Karunasagar, Iddya

2003-01-01

To study the incidence of Vibrio parahaemolyticus in seafoods, water and sediment by molecular techniques vs conventional microbiological methods. Of 86 samples analysed, 28 recorded positive for V. parahaemolyticus by conventional microbiological method, while 53 were positive by the toxR-targeted PCR, performed directly on enrichment broth lysates. While one sample of molluscan shellfish was positive for tdh gene, trh gene was detected in three enrichment broths of molluscan shellfish. Direct application of PCR to enrichment broths will be useful for the rapid and sensitive detection of potentially pathogenic strains of V. parahemolyticus in seafoods. Vibrio parahaemolyticus is an important human pathogen responsible for food-borne gastroenteritis world-wide. As, both pathogenic and non-pathogenic strains of V. parahaemolyticus exist in the seafood, application of PCR specific for the virulence genes (tdh & trh) will help in detection of pathogenic strains of V. parahaemolyticus and consequently reduce the risk of food-borne illness.

Fast learning method for convolutional neural networks using extreme learning machine and its application to lane detection.

PubMed

Kim, Jihun; Kim, Jonghong; Jang, Gil-Jin; Lee, Minho

2017-03-01

Deep learning has received significant attention recently as a promising solution to many problems in the area of artificial intelligence. Among several deep learning architectures, convolutional neural networks (CNNs) demonstrate superior performance when compared to other machine learning methods in the applications of object detection and recognition. We use a CNN for image enhancement and the detection of driving lanes on motorways. In general, the process of lane detection consists of edge extraction and line detection. A CNN can be used to enhance the input images before lane detection by excluding noise and obstacles that are irrelevant to the edge detection result. However, training conventional CNNs requires considerable computation and a big dataset. Therefore, we suggest a new learning algorithm for CNNs using an extreme learning machine (ELM). The ELM is a fast learning method used to calculate network weights between output and hidden layers in a single iteration and thus, can dramatically reduce learning time while producing accurate results with minimal training data. A conventional ELM can be applied to networks with a single hidden layer; as such, we propose a stacked ELM architecture in the CNN framework. Further, we modify the backpropagation algorithm to find the targets of hidden layers and effectively learn network weights while maintaining performance. Experimental results confirm that the proposed method is effective in reducing learning time and improving performance. Copyright © 2016 Elsevier Ltd. All rights reserved.

Rapid detection of Shigella and enteroinvasive Escherichia coli in produce enrichments by a conventional multiplex PCR assay.

PubMed

Binet, Rachel; Deer, Deanne M; Uhlfelder, Samantha J

2014-06-01

Faster detection of contaminated foods can prevent adulterated foods from being consumed and minimize the risk of an outbreak of foodborne illness. A sensitive molecular detection method is especially important for Shigella because ingestion of as few as 10 of these bacterial pathogens can cause disease. The objectives of this study were to compare the ability of four DNA extraction methods to detect Shigella in six types of produce, post-enrichment, and to evaluate a new and rapid conventional multiplex assay that targets the Shigella ipaH, virB and mxiC virulence genes. This assay can detect less than two Shigella cells in pure culture, even when the pathogen is mixed with background microflora, and it can also differentiate natural Shigella strains from a control strain and eliminate false positive results due to accidental laboratory contamination. The four DNA extraction methods (boiling, PrepMan Ultra [Applied Biosystems], InstaGene Matrix [Bio-Rad], DNeasy Tissue kit [Qiagen]) detected 1.6 × 10(3)Shigella CFU/ml post-enrichment, requiring ∼18 doublings to one cell in 25 g of produce pre-enrichment. Lower sensitivity was obtained, depending on produce type and extraction method. The InstaGene Matrix was the most consistent and sensitive and the multiplex assay accurately detected Shigella in less than 90 min, outperforming, to the best of our knowledge, molecular assays currently in place for this pathogen. Published by Elsevier Ltd.

Evaluation of nutrient agar for the culture of Mycobacterium tuberculosis using the microcolony detection method.

PubMed

Satti, L; Abbasi, S; Faiz, U

2012-07-01

We evaluated nutrient agar using the microcolony detection method for the recovery of Mycobacterium tuberculosis on 37 acid-fast bacilli (AFB) positive sputum specimens, and compared it with conventional Löwenstein-Jensen (LJ) medium. Nutrient agar detected 35 isolates compared to 34 on LJ medium. The mean time to detection of mycobacteria on nutrient agar and LJ medium was respectively 9.6 and 21.4 days. The contamination rate on nutrient agar and LJ medium was respectively 5.4% and 2.7%. Nutrient agar detects M. tuberculosis more rapidly than LJ medium, and could be an economical, rapid culture method in resource-poor settings, provided our findings are confirmed by further studies.

Real-time fluorescence loop mediated isothermal amplification for the diagnosis of malaria.

PubMed

Lucchi, Naomi W; Demas, Allison; Narayanan, Jothikumar; Sumari, Deborah; Kabanywanyi, Abdunoor; Kachur, S Patrick; Barnwell, John W; Udhayakumar, Venkatachalam

2010-10-29

Molecular diagnostic methods can complement existing tools to improve the diagnosis of malaria. However, they require good laboratory infrastructure thereby restricting their use to reference laboratories and research studies. Therefore, adopting molecular tools for routine use in malaria endemic countries will require simpler molecular platforms. The recently developed loop-mediated isothermal amplification (LAMP) method is relatively simple and can be improved for better use in endemic countries. In this study, we attempted to improve this method for malaria diagnosis by using a simple and portable device capable of performing both the amplification and detection (by fluorescence) of LAMP in one platform. We refer to this as the RealAmp method. Published genus-specific primers were used to test the utility of this method. DNA derived from different species of malaria parasites was used for the initial characterization. Clinical samples of P. falciparum were used to determine the sensitivity and specificity of this system compared to microscopy and a nested PCR method. Additionally, directly boiled parasite preparations were compared with a conventional DNA isolation method. The RealAmp method was found to be simple and allowed real-time detection of DNA amplification. The time to amplification varied but was generally less than 60 minutes. All human-infecting Plasmodium species were detected. The sensitivity and specificity of RealAmp in detecting P. falciparum was 96.7% and 91.7% respectively, compared to microscopy and 98.9% and 100% respectively, compared to a standard nested PCR method. In addition, this method consistently detected P. falciparum from directly boiled blood samples. This RealAmp method has great potential as a field usable molecular tool for diagnosis of malaria. This tool can provide an alternative to conventional PCR based diagnostic methods for field use in clinical and operational programs.

The Changing Role of the Clinical Microbiology Laboratory in Defining Resistance in Gram-negatives.

PubMed

Endimiani, Andrea; Jacobs, Michael R

2016-06-01

The evolution of resistance in Gram-negatives has challenged the clinical microbiology laboratory to implement new methods for their detection. Multidrug-resistant strains present major challenges to conventional and new detection methods. More rapid pathogen identification and antimicrobial susceptibility testing have been developed for use directly on specimens, including fluorescence in situ hybridization tests, automated polymerase chain reaction systems, microarrays, mass spectroscopy, next-generation sequencing, and microfluidics. Review of these methods shows the advances that have been made in rapid detection of resistance in cultures, but limited progress in direct detection from specimens. Copyright © 2016 Elsevier Inc. All rights reserved.

A further study to investigate the detection and enhancement of latent fingerprints using visible absorption and luminescence chemical imaging.

PubMed

Payne, Gemma; Reedy, Brian; Lennard, Chris; Comber, Bruce; Exline, David; Roux, Claude

2005-05-28

This study investigated the application of chemical imaging to the detection of latent fingerprints using the Condor macroscopic chemical imaging system (ChemImage Corp., Pittsburgh, USA). Methods were developed and optimised for the visualisation of untreated latent fingerprints and fingerprints processed with DFO, ninhydrin, cyanoacrylate, and cyanoacrylate plus rhodamine 6G stain. The results obtained with chemical imaging were compared to the detection achieved using conventional imaging techniques. The Condor significantly improved the detection of many prints, especially those that might be considered poor quality or borderline prints. Prints on newspaper treated with ninhydrin and DFO, and prints on white and yellow paper treated with ninhydrin, benefited the most from chemical imaging detection. In many cases, fingerprints undetectable using conventional imaging techniques could be visualised with chemical imaging. Ridge detail from untreated prints on yellow paper was also detected using the Condor. When prints of high quality were examined, both detection techniques produced quality results. The results of this project demonstrate that chemical imaging offers advantages over conventional visualisation techniques when examining latent fingerprints, especially those that would be considered difficult, such as weak prints or prints on surfaces that produce highly luminescent backgrounds. Standard testing procedures for the detection and enhancement of fingerprints by chemical imaging are presented and discussed.

Theoretical Analysis of Penalized Maximum-Likelihood Patlak Parametric Image Reconstruction in Dynamic PET for Lesion Detection.

PubMed

Yang, Li; Wang, Guobao; Qi, Jinyi

2016-04-01

Detecting cancerous lesions is a major clinical application of emission tomography. In a previous work, we studied penalized maximum-likelihood (PML) image reconstruction for lesion detection in static PET. Here we extend our theoretical analysis of static PET reconstruction to dynamic PET. We study both the conventional indirect reconstruction and direct reconstruction for Patlak parametric image estimation. In indirect reconstruction, Patlak parametric images are generated by first reconstructing a sequence of dynamic PET images, and then performing Patlak analysis on the time activity curves (TACs) pixel-by-pixel. In direct reconstruction, Patlak parametric images are estimated directly from raw sinogram data by incorporating the Patlak model into the image reconstruction procedure. PML reconstruction is used in both the indirect and direct reconstruction methods. We use a channelized Hotelling observer (CHO) to assess lesion detectability in Patlak parametric images. Simplified expressions for evaluating the lesion detectability have been derived and applied to the selection of the regularization parameter value to maximize detection performance. The proposed method is validated using computer-based Monte Carlo simulations. Good agreements between the theoretical predictions and the Monte Carlo results are observed. Both theoretical predictions and Monte Carlo simulation results show the benefit of the indirect and direct methods under optimized regularization parameters in dynamic PET reconstruction for lesion detection, when compared with the conventional static PET reconstruction.

Detection of foodborne pathogens using microarray technology

USDA-ARS?s Scientific Manuscript database

Assays based on the polymerase chain reaction (PCR) are now accepted methods for rapidly confirming the presence or absence of specific pathogens in foods and other types of samples. Conventional PCR requires the use of agarose gel electrophoresis to detect the PCR product; whereas, real-time PCR c...

Remote sensing change detection methods to track deforestation and growth in threatened rainforests in Madre de Dios, Peru

USGS Publications Warehouse

Shermeyer, Jacob S.; Haack, Barry N.

2015-01-01

Two forestry-change detection methods are described, compared, and contrasted for estimating deforestation and growth in threatened forests in southern Peru from 2000 to 2010. The methods used in this study rely on freely available data, including atmospherically corrected Landsat 5 Thematic Mapper and Moderate Resolution Imaging Spectroradiometer (MODIS) vegetation continuous fields (VCF). The two methods include a conventional supervised signature extraction method and a unique self-calibrating method called MODIS VCF guided forest/nonforest (FNF) masking. The process chain for each of these methods includes a threshold classification of MODIS VCF, training data or signature extraction, signature evaluation, k-nearest neighbor classification, analyst-guided reclassification, and postclassification image differencing to generate forest change maps. Comparisons of all methods were based on an accuracy assessment using 500 validation pixels. Results of this accuracy assessment indicate that FNF masking had a 5% higher overall accuracy and was superior to conventional supervised classification when estimating forest change. Both methods succeeded in classifying persistently forested and nonforested areas, and both had limitations when classifying forest change.

Detecting isotopic ratio outliers

NASA Astrophysics Data System (ADS)

Bayne, C. K.; Smith, D. H.

An alternative method is proposed for improving isotopic ratio estimates. This method mathematically models pulse-count data and uses iterative reweighted Poisson regression to estimate model parameters to calculate the isotopic ratios. This computer-oriented approach provides theoretically better methods than conventional techniques to establish error limits and to identify outliers.

A computerized scheme for lung nodule detection in multiprojection chest radiography

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

Guo Wei; Li Qiang; Boyce, Sarah J.

2012-04-15

Purpose: Our previous study indicated that multiprojection chest radiography could significantly improve radiologists' performance for lung nodule detection in clinical practice. In this study, the authors further verify that multiprojection chest radiography can greatly improve the performance of a computer-aided diagnostic (CAD) scheme. Methods: Our database consisted of 59 subjects, including 43 subjects with 45 nodules and 16 subjects without nodules. The 45 nodules included 7 real and 38 simulated ones. The authors developed a conventional CAD scheme and a new fusion CAD scheme to detect lung nodules. The conventional CAD scheme consisted of four steps for (1) identification ofmore » initial nodule candidates inside lungs, (2) nodule candidate segmentation based on dynamic programming, (3) extraction of 33 features from nodule candidates, and (4) false positive reduction using a piecewise linear classifier. The conventional CAD scheme processed each of the three projection images of a subject independently and discarded the correlation information between the three images. The fusion CAD scheme included the four steps in the conventional CAD scheme and two additional steps for (5) registration of all candidates in the three images of a subject, and (6) integration of correlation information between the registered candidates in the three images. The integration step retained all candidates detected at least twice in the three images of a subject and removed those detected only once in the three images as false positives. A leave-one-subject-out testing method was used for evaluation of the performance levels of the two CAD schemes. Results: At the sensitivities of 70%, 65%, and 60%, our conventional CAD scheme reported 14.7, 11.3, and 8.6 false positives per image, respectively, whereas our fusion CAD scheme reported 3.9, 1.9, and 1.2 false positives per image, and 5.5, 2.8, and 1.7 false positives per patient, respectively. The low performance of the conventional CAD scheme may be attributed to the high noise level in chest radiography, and the small size and low contrast of most nodules. Conclusions: This study indicated that the fusion of correlation information in multiprojection chest radiography can markedly improve the performance of CAD scheme for lung nodule detection.« less

Real-time line-width measurements: a new feature for reticle inspection systems

NASA Astrophysics Data System (ADS)

Eran, Yair; Greenberg, Gad; Joseph, Amnon; Lustig, Cornel; Mizrahi, Eyal

1997-07-01

The significance of line width control in mask production has become greater with the lessening of defect size. There are two conventional methods used for controlling line widths dimensions which employed in the manufacturing of masks for sub micron devices. These two methods are the critical dimensions (CD) measurement and the detection of edge defects. Achieving reliable and accurate control of line width errors is one of the most challenging tasks in mask production. Neither of the two methods cited above (namely CD measurement and the detection of edge defects) guarantees the detection of line width errors with good sensitivity over the whole mask area. This stems from the fact that CD measurement provides only statistical data on the mask features whereas applying edge defect detection method checks defects on each edge by itself, and does not supply information on the combined result of error detection on two adjacent edges. For example, a combination of a small edge defect together with a CD non- uniformity which are both within the allowed tolerance, may yield a significant line width error, which will not be detected using the conventional methods (see figure 1). A new approach for the detection of line width errors which overcomes this difficulty is presented. Based on this approach, a new sensitive line width error detector was developed and added to Orbot's RT-8000 die-to-database reticle inspection system. This innovative detector operates continuously during the mask inspection process and scans (inspects) the entire area of the reticle for line width errors. The detection is based on a comparison of measured line width that are taken on both the design database and the scanned image of the reticle. In section 2, the motivation for developing this new detector is presented. The section covers an analysis of various defect types, which are difficult to detect using conventional edge detection methods or, alternatively, CD measurements. In section 3, the basic concept of the new approach is introduced together with a description of the new detector and its characteristics. In section 4, the calibration process that took place in order to achieve reliable and repeatable line width measurements is presented. The description of an experiments conducted in order to evaluate the sensitivity of the new detector is given in section 5, followed by a report of the results of this evaluation. The conclusions are presented in section 6.

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

Lee, Jaesun, E-mail: jaesun@pusan.ac.kr, E-mail: jpp@pusan.ac.kr; Park, Junpil, E-mail: jaesun@pusan.ac.kr, E-mail: jpp@pusan.ac.kr; Cho, Younho, E-mail: mechcyh@pusan.ac.kr

The nuclear power plant inspection is very important for the safety issue. However due to some radiation and geometric problems, the detection of CRDM(Control Rod Drive Mechanism) can be very difficult by using conventional Ultrasonic Testing method. Also the shrink fit boundary condition can also be an obstacle for the inspection in this paper, instead of conventional Ultrasonic Testing, guided wave was used for the detection of some complicated structures. The CRDM nozzle was installed in reactor head with perfect shrink fit condition by using stainless steel. The wave amplitude distribution on the circumferential direction was calculated with various boundarymore » conditions and the experimental result shows a possibility of the defect detection on J-groove weld.« less

Tracing the conformational changes in BSA using FRET with environmentally-sensitive squaraine probes

NASA Astrophysics Data System (ADS)

Govor, Iryna V.; Tatarets, Anatoliy L.; Obukhova, Olena M.; Terpetschnig, Ewald A.; Gellerman, Gary; Patsenker, Leonid D.

2016-06-01

A new potential method of detecting the conformational changes in hydrophobic proteins such as bovine serum albumin (BSA) is introduced. The method is based on the change in the Förster resonance energy transfer (FRET) efficiency between protein-sensitive fluorescent probes. As compared to conventional FRET based methods, in this new approach the donor and acceptor dyes are not covalently linked to protein molecules. Performance of the new method is demonstrated using the protein-sensitive squaraine probes Square-634 (donor) and Square-685 (acceptor) to detect the urea-induced conformational changes of BSA. The FRET efficiency between these probes can be considered a more sensitive parameter to trace protein unfolding as compared to the changes in fluorescence intensity of each of these probes. Addition of urea followed by BSA unfolding causes a noticeable decrease in the emission intensities of these probes (factor of 5.6 for Square-634 and 3.0 for Square-685), and the FRET efficiency changes by a factor of up to 17. Compared to the conventional method the new approach therefore demonstrates to be a more sensitive way to detect the conformational changes in BSA.

An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection.

PubMed

Pitz, Amanda de Faveri; Koishi, Andrea Cristine; Tavares, Eliandro Reis; Andrade, Fábio Goulart de; Loth, Eduardo Alexandre; Gandra, Rinaldo Ferreira; Venancio, Emerson José

2013-01-01

Herein, we report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR) assay for the detection of Paracoccidioides brasiliensis. We developed the OTsn-PCR assay for the detection of P. brasiliensis in clinical specimens and compared it with other PCR methods. The OTsn-PCR assay was positive for all clinical samples, and the detection limit was better or equivalent to the other nested or semi-nested PCR methods for P. brasiliensis detection. The OTsn-PCR assay described in this paper has a detection limit similar to other reactions for the molecular detection of P. brasiliensis, but this approach is faster and less prone to contamination than other conventional nested or semi-nested PCR assays.

Loop-mediated isothermal amplification assay targeting the mpb70 gene for rapid differential detection of Mycobacterium bovis.

PubMed

Zhang, Hui; Wang, Zhen; Cao, Xudong; Wang, Zhengrong; Sheng, Jinliang; Wang, Yong; Zhang, Jing; Li, Zhiqiang; Gu, Xinli; Chen, Chuangfu

2016-11-01

Loop-mediated isothermal amplification (LAMP) is a highly sensitive, rapid, cost-effective nucleic acid amplification method. Tuberculosis (TB) is widely popular in the world and it is difficult to cure. The fundamental treatment is to clear the types of TB pathogens such as Mycobacterium bovis (M. bovis), Mycobacterium tuberculosis (M. tuberculosis). In order to detect and diagnose TB early, we constructed the differential diagnostic method of TB. In this study, we used LAMP for detection of M. bovis, based on amplification of the mpb70 gene which is a unique gene in M. bovis strain. The LAMP assay was able to detect only seven copies of the gene per reaction, whereas for the conventional PCR, it was 70 copies. The LAMP was evaluated for its specificity using six strains of five Mycobacterium species and 18 related non-Mycobacterium microorganism strains as controls. The target three Mycobacterium strains were all amplified, and no cross-reaction was found with 18 non-Mycobacterium microorganism strains. TB was detected by two methods, LAMP and conventional PCR (based on mpb70 gene); the positive rates of the two methods were 9.55 and 7.01 %, respectively. Our results indicate that the LAMP method should be a potential tool with high convenience, rapidity, sensitivity and specificity for the diagnosis of TB caused by M. bovis. Most importance is that the use of LAMP as diagnostic method in association with diagnostic tests based on mpb70 gene would allow the differentiation between M. bovis and other Mycobacterium in humans or animals. The LAMP method is actually in order to detect human TB, and it can be used for differential diagnosis in this paper.

A new technique to expose the hypopharyngeal space: The modified Killian's method.

PubMed

Sakai, Akihiro; Okami, Kenji; Sugimoto, Ryousuke; Ebisumoto, Koji; Yamamoto, Hikaru; Maki, Daisuke; Saito, Kosuke; Iida, Masahiro

2014-04-01

Recent remarkable progress in endoscopic technology has enabled the detection of superficial cancers that were undetectable in the past. However, even though advanced endoscopic technology can detect early lesions, it is useless unless it can provide wide exposure of an area. By modifying the Killian position, it is possible to observe a wider range of the hypopharyngeal space than is possible with conventional head positions. We report a revolutionary method that uses a new head position to widely open the hypopharynx. The technique is named "the Modified Killian's method." The patient is initially placed in the Killian position and then bent further forward from the original position (i.e., the modified Killian position). While in this position, the patient's head is turned and the Valsalva maneuver is applied. These additional maneuvers constitute the Modified Killian's method and widely expands the hypopharyngeal space. The conventional head position cannot open the hypopharyngeal space sufficiently; however, the Modified Killian's method opens the hypopharyngeal space very widely. The Modified Killian's method enables observation of the entire circumference of the hypopharyngeal space and the cervical esophageal entry. The Modified Killian's method may become an indispensable technique for observing the hypopharynx and detecting hypopharyngeal cancers. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

Distribution majorization of corner points by reinforcement learning for moving object detection

NASA Astrophysics Data System (ADS)

Wu, Hao; Yu, Hao; Zhou, Dongxiang; Cheng, Yongqiang

2018-04-01

Corner points play an important role in moving object detection, especially in the case of free-moving camera. Corner points provide more accurate information than other pixels and reduce the computation which is unnecessary. Previous works only use intensity information to locate the corner points, however, the information that former and the last frames provided also can be used. We utilize the information to focus on more valuable area and ignore the invaluable area. The proposed algorithm is based on reinforcement learning, which regards the detection of corner points as a Markov process. In the Markov model, the video to be detected is regarded as environment, the selections of blocks for one corner point are regarded as actions and the performance of detection is regarded as state. Corner points are assigned to be the blocks which are seperated from original whole image. Experimentally, we select a conventional method which uses marching and Random Sample Consensus algorithm to obtain objects as the main framework and utilize our algorithm to improve the result. The comparison between the conventional method and the same one with our algorithm show that our algorithm reduce 70% of the false detection.

Novel primers and PCR protocols for the specific detection and quantification of Sphingobium suberifaciens in situ

USDA-ARS?s Scientific Manuscript database

The pathogen causing corky root on lettuce, Sphingobium suberifaciens, is recalcitrant to standard epidemiological methods. Primers were selected from 16S rDNA sequences useful for the specific detection and quantification of S. suberifaciens. Conventional (PCR) and quantitative (qPCR) PCR protocols...

Applicability of a Conservative Margin Approach for Assessing NDE Flaw Detectability

NASA Technical Reports Server (NTRS)

Koshti, ajay M.

2007-01-01

Nondestructive Evaluation (NDE) procedures are required to detect flaws in structures with a high percentage detectability and high confidence. Conventional Probability of Detection (POD) methods are statistical in nature and require detection data from a relatively large number of flaw specimens. In many circumstances, due to the high cost and long lead time, it is impractical to build the large set of flaw specimens that is required by the conventional POD methodology. Therefore, in such situations it is desirable to have a flaw detectability estimation approach that allows for a reduced number of flaw specimens but provides a high degree of confidence in establishing the flaw detectability size. This paper presents an alternative approach called the conservative margin approach (CMA). To investigate the applicability of the CMA approach, flaw detectability sizes determined by the CMA and POD approaches have been compared on actual datasets. The results of these comparisons are presented and the applicability of the CMA approach is discussed.

Calculating the ankylosing spondylitis disease activity score if the conventional c-reactive protein level is below the limit of detection or if high-sensitivity c-reactive protein is used: an analysis in the DESIR cohort.

PubMed

Machado, Pedro; Navarro-Compán, Victoria; Landewé, Robert; van Gaalen, Floris A; Roux, Christian; van der Heijde, Désirée

2015-02-01

The Ankylosing Spondylitis Disease Activity Score (ASDAS) is a composite measure of disease activity in axial spondyloarthritis. The aims of this study were to determine the most appropriate method for calculating the ASDAS using the C-reactive protein (CRP) level when the conventional CRP level was below the limit of detection, to determine how low CRP values obtained by high-sensitivity CRP (hsCRP) measurement influence ASDAS-CRP results, and to test agreement between different ASDAS formulae. Patients with axial spondyloarthritis who had a conventional CRP level below the limit of detection (5 mg/liter) were selected (n = 257). The ASDAS–conventional CRP with 11 different imputations for the conventional CRP value (range 0–5 mg/liter, at 0.5-mg/liter intervals) was calculated. The ASDAS-hsCRP and ASDAS using the erythrocyte sedimentation rate (ESR) were also calculated. Agreement between the ASDAS formulae was tested. The ASDAS-hsCRP showed better agreement with the ASDAS-CRP calculated using the conventional CRP imputation values of 1.5 and 2.0 mg/liter and with the ASDAS-ESR than with other imputed formulae. Disagreement occurred mainly in lower disease activity states (inactive/moderate disease activity). When the CRP value was <2 mg/liter, the resulting ASDAS-CRP scores may have been inappropriately low. When the conventional CRP level is below the limit of detection or when the hsCRP level is <2 mg/liter, the constant value of 2 mg/liter should be used to calculate the ASDAS-CRP score. There is good agreement between the ASDAS-hsCRP and ASDAS-ESR; however, formulae are not interchangeable.

An evaluation of computer-aided disproportionality analysis for post-marketing signal detection.

PubMed

Lehman, H P; Chen, J; Gould, A L; Kassekert, R; Beninger, P R; Carney, R; Goldberg, M; Goss, M A; Kidos, K; Sharrar, R G; Shields, K; Sweet, A; Wiholm, B E; Honig, P K

2007-08-01

To understand the value of computer-aided disproportionality analysis (DA) in relation to current pharmacovigilance signal detection methods, four products were retrospectively evaluated by applying an empirical Bayes method to Merck's post-marketing safety database. Findings were compared with the prior detection of labeled post-marketing adverse events. Disproportionality ratios (empirical Bayes geometric mean lower 95% bounds for the posterior distribution (EBGM05)) were generated for product-event pairs. Overall (1993-2004 data, EBGM05> or =2, individual terms) results of signal detection using DA compared to standard methods were sensitivity, 31.1%; specificity, 95.3%; and positive predictive value, 19.9%. Using groupings of synonymous labeled terms, sensitivity improved (40.9%). More of the adverse events detected by both methods were detected earlier using DA and grouped (versus individual) terms. With 1939-2004 data, diagnostic properties were similar to those from 1993 to 2004. DA methods using Merck's safety database demonstrate sufficient sensitivity and specificity to be considered for use as an adjunct to conventional signal detection methods.

Development and application of a rapid and visual loop-mediated isothermal amplification for the detection of Sporisorium scitamineum in sugarcane

PubMed Central

Su, Yachun; Yang, Yuting; Peng, Qiong; Zhou, Dinggang; Chen, Yun; Wang, Zhuqing; Xu, Liping; Que, Youxiong

2016-01-01

Smut is a fungal disease with widespread prevalence in sugarcane planting areas. Early detection and proper identification of Sporisorium scitamineum are essential in smut management practices. In the present study, four specific primers targeting the core effector Pep1 gene of S. scitamineum were designed. Optimal concentrations of Mg2+, primer and Bst DNA polymerase, the three important components of the loop-mediated isothermal amplification (LAMP) reaction system, were screened using a single factor experiment method and the L16(45) orthogonal experimental design. Hence, a LAMP system suitable for detection of S. scitamineum was established. High specificity of the LAMP method was confirmed by the assay of S. scitamineum, Fusarium moniliforme, Pestalotia ginkgo, Helminthospcrium sacchari, Fusarium oxysporum and endophytes of Yacheng05-179 and ROC22. The sensitivity of the LAMP method was equal to that of the conventional PCR targeting Pep1 gene and was 100 times higher than that of the conventional PCR assay targeting bE gene in S. scitamineum. The results suggest that this novel LAMP system has strong specificity and high sensitivity. This method not only provides technological support for the epidemic monitoring of sugarcane smut, but also provides a good case for development of similar detection technology for other plant pathogens. PMID:27035751

Location detection and tracking of moving targets by a 2D IR-UWB radar system.

PubMed

Nguyen, Van-Han; Pyun, Jae-Young

2015-03-19

In indoor environments, the Global Positioning System (GPS) and long-range tracking radar systems are not optimal, because of signal propagation limitations in the indoor environment. In recent years, the use of ultra-wide band (UWB) technology has become a possible solution for object detection, localization and tracking in indoor environments, because of its high range resolution, compact size and low cost. This paper presents improved target detection and tracking techniques for moving objects with impulse-radio UWB (IR-UWB) radar in a short-range indoor area. This is achieved through signal-processing steps, such as clutter reduction, target detection, target localization and tracking. In this paper, we introduce a new combination consisting of our proposed signal-processing procedures. In the clutter-reduction step, a filtering method that uses a Kalman filter (KF) is proposed. Then, in the target detection step, a modification of the conventional CLEAN algorithm which is used to estimate the impulse response from observation region is applied for the advanced elimination of false alarms. Then, the output is fed into the target localization and tracking step, in which the target location and trajectory are determined and tracked by using unscented KF in two-dimensional coordinates. In each step, the proposed methods are compared to conventional methods to demonstrate the differences in performance. The experiments are carried out using actual IR-UWB radar under different scenarios. The results verify that the proposed methods can improve the probability and efficiency of target detection and tracking.

Characterization by culture-dependent and culture-independent methods of the bacterial population of suckling-lamb packaged in different atmospheres.

PubMed

Osés, Sandra M; Diez, Ana M; Melero, Beatriz; Luning, Pieternel A; Jaime, Isabel; Rovira, Jordi

2013-12-01

This study offers insight into the dynamics of bacterial populations in fresh cuts of suckling lamb under four different atmospheric conditions: air (A), and three Modified Atmosphere Packaging (MAP) environments, 15%O2/30%CO2/55%N2 (C, commercial), 70%O2/30%CO2 (O), and 15%O2/85%CO2 (H) for 18 days. Microbial analyses by both conventional methods and PCR-DGGE were performed. Controversial and surprising results emerged from comparing both methods in relation to the genus Pseudomonas. Thus, conventional methods detected the presence of high numbers of Pseudomonas colonies, although PCR-DGGE only detected this genus in air-packaged samples. PCR-DGGE detected higher microbial diversity in the control samples (A) than in the modified atmospheres (C, O, H), having atmosphere H the fewest number of species. Brochothrix thermosphacta, LAB (Carnobacterium divergens and Lactobacillus sakei), and Escherichia spp. were detected in all the atmospheres throughout storage. Moreover, previously undescribed bacteria from lamb meat such as Enterobacter hormaechei, Staphylococcus equorum and Jeotgalicoccus spp. were also isolated in this study by DGGE. Additionally, qPCR analysis was used to detect and characterize strains of Escherichia coli. Virulence genes (stx1, stx2 and eae) were detected throughout storage in 97% of the samples. A high CO2 atmosphere was the most effective packaging combination doubling storage time in comparison with commercial atmosphere. Copyright © 2013 Elsevier Ltd. All rights reserved.

Rapid and sensitive detection of mink circovirus by recombinase polymerase amplification.

PubMed

Ge, Junwei; Shi, Yunjia; Cui, Xingyang; Gu, Shanshan; Zhao, Lili; Chen, Hongyan

2018-06-01

To date, the pathogenic role of mink circovirus (MiCV) remains unclear, and its prevalence and economic importance are unknown. Therefore, a rapid and sensitive molecular diagnosis is necessary for disease management and epidemiological surveillance. However, only PCR methods can identify MiCV infection at present. In this study, we developed a nested PCR and established a novel recombinase polymerase amplification (RPA) assay for MiCV detection. Sensitivity analysis showed that the detection limit of nested PCR and RPA assay was 10 1 copies/reaction, and these methods were more sensitive than conventional PCR, which has a detection limit of 10 5 copies/reaction. The RPA assay had no cross-reactivity with other related viral pathogens, and amplification was completed in less than 20 min with a simple device. Further assessment of clinical samples showed that the two assays were accurate in identifying positive and negative conventional PCR samples. The detection rate of MiCV by the RPA assay in clinical samples was 38.09%, which was 97% consistent with that by the nested PCR. The developed nested PCR is a highly sensitive tool for practical use, and the RPA assay is a simple, sensitive, and potential alternative method for rapid and accurate MiCV diagnosis. Copyright © 2018 Elsevier B.V. All rights reserved.

Diagnostic performance of blood culture bottles for vitreous culture compared to conventional microbiological cultures in patients with suspected endophthalmitis.

PubMed

Kehrmann, Jan; Chapot, Valerie; Buer, Jan; Rating, Philipp; Bornfeld, Norbert; Steinmann, Joerg

2018-05-01

The purpose of this investigation was to evaluate the performance of blood culture bottles in comparison to conventional microbiological culture techniques in detecting causative microorganisms of endophthalmitis and to determine their anti-infective susceptibility profiles. All consecutive cases with clinically suspected endophthalmitis in a university-based ophthalmology department between January 2009 and December 2016 were analysed in this retrospective comparative case series. Samples from 247 patients with suspected endophthalmitis underwent microbiological diagnostic work-up. All three culture methods were performed from 140 vitreous specimens. Vitreous fluid specimens were inoculated in blood culture bottles, aerobic and anaerobic broth solutions, and on solid media. Anti-infective susceptibility profiles were evaluated by semi-automated methods and/or gradient diffusion methods. Microorganisms were grown in 82 of 140 specimens for which all methods were performed (59%). Microorganisms were more frequently grown from blood culture bottles (55%) compared to broth solution (45%, p = 0.007) and solid media (33%, p < 0.0001). Considerable differences in the performance among culture media were detected for fungal pathogens. All grown fungi were detected by blood culture bottles (11 of 11, 100%). Broth solution recovered 64% and solid media 46% of grown fungi. No Gram-positive bacterium was resistant to vancomycin and all Gram-negative pathogens except for one isolate were susceptible to third-generation cephalosporins. In suspected endophthalmitis patients, blood culture bottles have a higher overall pathogen detection rate from vitreous fluid compared to conventional microbiological media, especially for fungi. The initial intravitreal antibiotic therapy with vancomycin plus third-generation cephalosporins appears to be an appropriate treatment approach for bacterial endophthalmitis.

Novel Method For Low-Rate Ddos Attack Detection

NASA Astrophysics Data System (ADS)

Chistokhodova, A. A.; Sidorov, I. D.

2018-05-01

The relevance of the work is associated with an increasing number of advanced types of DDoS attacks, in particular, low-rate HTTP-flood. Last year, the power and complexity of such attacks increased significantly. The article is devoted to the analysis of DDoS attacks detecting methods and their modifications with the purpose of increasing the accuracy of DDoS attack detection. The article details low-rate attacks features in comparison with conventional DDoS attacks. During the analysis, significant shortcomings of the available method for detecting low-rate DDoS attacks were found. Thus, the result of the study is an informal description of a new method for detecting low-rate denial-of-service attacks. The architecture of the stand for approbation of the method is developed. At the current stage of the study, it is possible to improve the efficiency of an already existing method by using a classifier with memory, as well as additional information.

Flow cytometric detection method for DNA samples

DOEpatents

Nasarabadi, Shanavaz [Livermore, CA; Langlois, Richard G [Livermore, CA; Venkateswaran, Kodumudi S [Round Rock, TX

2011-07-05

Disclosed herein are two methods for rapid multiplex analysis to determine the presence and identity of target DNA sequences within a DNA sample. Both methods use reporting DNA sequences, e.g., modified conventional Taqman.RTM. probes, to combine multiplex PCR amplification with microsphere-based hybridization using flow cytometry means of detection. Real-time PCR detection can also be incorporated. The first method uses a cyanine dye, such as, Cy3.TM., as the reporter linked to the 5' end of a reporting DNA sequence. The second method positions a reporter dye, e.g., FAM.TM. on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA.TM., on the 5' end.

Flow cytometric detection method for DNA samples

DOEpatents

Nasarabadi, Shanavaz [Livermore, CA; Langlois, Richard G [Livermore, CA; Venkateswaran, Kodumudi S [Livermore, CA

2006-08-01

Disclosed herein are two methods for rapid multiplex analysis to determine the presence and identity of target DNA sequences within a DNA sample. Both methods use reporting DNA sequences, e.g., modified conventional Taqman.RTM. probes, to combine multiplex PCR amplification with microsphere-based hybridization using flow cytometry means of detection. Real-time PCR detection can also be incorporated. The first method uses a cyanine dye, such as, Cy3.TM., as the reporter linked to the 5' end of a reporting DNA sequence. The second method positions a reporter dye, e.g., FAM, on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA, on the 5' end.

Event specific qualitative and quantitative polymerase chain reaction detection of genetically modified MON863 maize based on the 5'-transgene integration sequence.

PubMed

Yang, Litao; Xu, Songci; Pan, Aihu; Yin, Changsong; Zhang, Kewei; Wang, Zhenying; Zhou, Zhigang; Zhang, Dabing

2005-11-30

Because of the genetically modified organisms (GMOs) labeling policies issued in many countries and areas, polymerase chain reaction (PCR) methods were developed for the execution of GMO labeling policies, such as screening, gene specific, construct specific, and event specific PCR detection methods, which have become a mainstay of GMOs detection. The event specific PCR detection method is the primary trend in GMOs detection because of its high specificity based on the flanking sequence of the exogenous integrant. This genetically modified maize, MON863, contains a Cry3Bb1 coding sequence that produces a protein with enhanced insecticidal activity against the coleopteran pest, corn rootworm. In this study, the 5'-integration junction sequence between the host plant DNA and the integrated gene construct of the genetically modified maize MON863 was revealed by means of thermal asymmetric interlaced-PCR, and the specific PCR primers and TaqMan probe were designed based upon the revealed 5'-integration junction sequence; the conventional qualitative PCR and quantitative TaqMan real-time PCR detection methods employing these primers and probes were successfully developed. In conventional qualitative PCR assay, the limit of detection (LOD) was 0.1% for MON863 in 100 ng of maize genomic DNA for one reaction. In the quantitative TaqMan real-time PCR assay, the LOD and the limit of quantification were eight and 80 haploid genome copies, respectively. In addition, three mixed maize samples with known MON863 contents were detected using the established real-time PCR systems, and the ideal results indicated that the established event specific real-time PCR detection systems were reliable, sensitive, and accurate.

Evaluation of the efficiency of three extraction conditions for the immunochemical detection of allergenic soy proteins in different food matrices.

PubMed

Amponsah, Amma; Nayak, Balunkeswar

2018-04-01

Recent studies have shown the need to improve soy allergen extraction using different extraction conditions to ensure more accurate results in allergen detection. This study investigated some of these extraction conditions to confirm that these methods, especially ultrasound-assisted extraction (UAE) and the use of Laemmli buffer instead of the conventional extraction with phosphate-buffered saline (PBS), could be helpful in improving the extraction step in allergen detection. Higher total soluble protein was obtained in all samples extracted with Laemmli buffer alone and in combination with ultrasound. For immunochemical detection of soy proteins by enzyme-linked immunosorbent assay (ELISA), comparable detection was observed in extracts from all extraction conditions in all commercial samples with the exception of table cracker and veggie burger, where significantly higher detection was seen in extracts from Laemmli buffer only. For the dry mix and cookie samples, the degree of soy protein detection with ELISA varied among the different extraction conditions, but overall, extraction with only Laemmli buffer showed higher detection. Laemmli buffer with conventional extraction and UAE may be better alternatives or additional extraction methods in soy allergen detection. Different food matrices performed differently (whether it was for the recovery of total proteins or detection by ELISA) under different extraction conditions. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Leak Detection and Location of Water Pipes Using Vibration Sensors and Modified ML Prefilter.

PubMed

Choi, Jihoon; Shin, Joonho; Song, Choonggeun; Han, Suyong; Park, Doo Il

2017-09-13

This paper proposes a new leak detection and location method based on vibration sensors and generalised cross-correlation techniques. Considering the estimation errors of the power spectral densities (PSDs) and the cross-spectral density (CSD), the proposed method employs a modified maximum-likelihood (ML) prefilter with a regularisation factor. We derive a theoretical variance of the time difference estimation error through summation in the discrete-frequency domain, and find the optimal regularisation factor that minimises the theoretical variance in practical water pipe channels. The proposed method is compared with conventional correlation-based techniques via numerical simulations using a water pipe channel model, and it is shown through field measurement that the proposed modified ML prefilter outperforms conventional prefilters for the generalised cross-correlation. In addition, we provide a formula to calculate the leak location using the time difference estimate when different types of pipes are connected.

Tuning fork enhanced interferometric photoacoustic spectroscopy: a new method for trace gas analysis

NASA Astrophysics Data System (ADS)

Köhring, M.; Pohlkötter, A.; Willer, U.; Angelmahr, M.; Schade, W.

2011-01-01

A photoacoustic trace gas sensor based on an optical read-out method of a quartz tuning fork is shown. Instead of conventional piezoelectric signal read-out, as applied in well-known quartz-enhanced photoacoustic spectroscopy (QEPAS), an interferometric read-out method for measurement of the tuning fork's oscillation is presented. To demonstrate the potential of the optical read-out of tuning forks in photoacoustics, a comparison between the performances of a sensor with interferometric read-out and conventional QEPAS with piezoelectric read-out is reported. The two sensors show similar characteristics. The detection limit (L) for the optical read-out is determined to be L opt=(2598±84) ppm (1 σ) compared to L elec=(2579±78) ppm (1 σ) for piezoelectric read-out. In both cases the detection limit is defined by the thermal noise of the tuning fork.

Leak Detection and Location of Water Pipes Using Vibration Sensors and Modified ML Prefilter

PubMed Central

Shin, Joonho; Song, Choonggeun; Han, Suyong; Park, Doo Il

2017-01-01

This paper proposes a new leak detection and location method based on vibration sensors and generalised cross-correlation techniques. Considering the estimation errors of the power spectral densities (PSDs) and the cross-spectral density (CSD), the proposed method employs a modified maximum-likelihood (ML) prefilter with a regularisation factor. We derive a theoretical variance of the time difference estimation error through summation in the discrete-frequency domain, and find the optimal regularisation factor that minimises the theoretical variance in practical water pipe channels. The proposed method is compared with conventional correlation-based techniques via numerical simulations using a water pipe channel model, and it is shown through field measurement that the proposed modified ML prefilter outperforms conventional prefilters for the generalised cross-correlation. In addition, we provide a formula to calculate the leak location using the time difference estimate when different types of pipes are connected. PMID:28902154

Comparison of Para-Selles Bailenger/Kop-Color Fumouze, Para-Selles-Iodésine/Kop-Color II Fumouze diagnostic kits with conventional microscopic methods in identifying intestinal parasitic diseases in Senegal.

PubMed

Sow, Doudou; Dieng, Yémou; Haouchine, Djamal; Niang, Khadim; Niang, Thiane; Sylla, Khadime; Tine, Roger Clément; Ndiaye, Magatte; Ndiaye, Jean Louis; Faye, Babacar; Faye, Omar; Gaye, Oumar; Dieng, Thérèse; Izri, Arezki

2017-09-01

In the context of controlling intestinal parasites, accurate diagnosis is essential. Our objective was to evaluate the performance of new diagnostic kits compared to conventional microscopic methods in identifying intestinal parasites. Faeces collected in rural area in Senegal were subjected to several detection techniques. Thus, the sensitivity, specificity, positive and negative predictive values of new diagnostic techniques were compared to conventional merthiolate-iodine-formalin, conventional Bailenger and modified Ritchie. Furthermore, the kappa coefficient was calculated to evaluate the correlation between the new kit and those of modified Ritchie. Out of the 117 patients examined, 102 presented with a parasite, or prevalence of 87.1%. The Fumouze techniques proved to be as effective as the conventional methods in detecting flagellates and helminths with sensitivities ranging from 97 to 100%. However, conventional techniques were slightly more sensitive in identifying Endolimax nana and Blastocystis hominis . The correlation was nearly perfect (k = 0.83 and 1), respectively between Bailenger Fumouze, Iodesine Fumouze and modified Ritchie in identifying helminths while it was just acceptable (k = 0.27 and 0.28) in identifying B. hominis . The modified Ritchie technique routinely used in our laboratory remains a good diagnostic tool. However, the use of kit techniques was interesting when reading the pellet after concentration and the Colour KOP staining was a considerable contribution to the diagnosis of the vegetative forms. Therefore, it would be interesting to determine the cost of a stool test using Fumouze kit techniques to provide the most cost effective way.

Rapid and sensitive detection of Lily symptomless virus by reverse transcription loop-mediated isothermal amplification.

PubMed

He, Xiangfeng; Xue, Fei; Xu, Shufa; Wang, Wenhe

2016-12-01

Lily symptomless virus (LSV) is one of the most prevalent viruses that infect lily plants worldwide. A rapid and sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of LSV, using two primer pairs that specifically amplified the conserved sequence of LSV coat protein. The optimum reaction conditions were as follows: 4mM MgCl 2 and 0.8M betaine with incubation at 64°C for 30min. The limit of detection of LSV from infected lily leaves was 10-fold higher for RT-LAMP than for conventional RT-PCR. Moreover, RT-LAMP detected LSV in not only symptomatic, but also in symptomless tissues of infected plants. These findings indicate that our RT-LAMP method for LSV can serve as a low-cost, simple, and rapid alternative to conventional detection assays. Copyright © 2016 Elsevier B.V. All rights reserved.

Enzymatic signal amplification for sensitive detection of intracellular antigens by flow cytometry.

PubMed

Karkmann, U; Radbruch, A; Hölzel, V; Scheffold, A

1999-11-19

Flow cytometry is the method of choice for the analysis of single cells with respect to the expression of specific antigens. Antigens can be detected with specific antibodies either on the cell surface or within the cells, after fixation and permeabilization of the cell membrane. Using conventional fluorochrome-labeled antibodies several thousand antigens are required for clear-cut separation of positive and negative cells. More sensitive reagents, e.g., magnetofluorescent liposomes conjugated to specific antibodies permit the detection of less than 200 molecules per cell but cannot be used for the detection of intracellular antigens. Here, we describe an enzymatic amplification technique (intracellular tyramine-based signal amplification, ITSA) for the sensitive cytometric analysis of intracellular cytokines by immunofluorescence. This approach results in a 10 to 15-fold improvement of the signal-to-noise ratio compared to conventional fluorochrome labeled antibodies and permits the detection of as few as 300-400 intracellular antigens per cell.

A novel quantitative real-time polymerase chain reaction method for detecting toxigenic Pasteurella multocida in nasal swabs from swine.

PubMed

Scherrer, Simone; Frei, Daniel; Wittenbrink, Max Michael

2016-12-01

Progressive atrophic rhinitis (PAR) in pigs is caused by toxigenic Pasteurella multocida. In Switzerland, PAR is monitored by selective culture of nasal swabs and subsequent polymerase chain reaction (PCR) screening of bacterial colonies for the P. multocida toxA gene. A panel of 203 nasal swabs from a recent PAR outbreak were used to evaluate a novel quantitative real-time PCR for toxigenic P. multocida in porcine nasal swabs. In comparison to the conventional PCR with a limit of detection of 100 genome equivalents per PCR reaction, the real-time PCR had a limit of detection of 10 genome equivalents. The real-time PCR detected toxA-positive P. multocida in 101 samples (49.8%), whereas the conventional PCR was less sensitive with 90 toxA-positive samples (44.3%). In comparison to the real-time PCR, 5.4% of the toxA-positive samples revealed unevaluable results by conventional PCR. The approach of culture-coupled toxA PCR for the monitoring of PAR in pigs is substantially improved by a novel quantitative real-time PCR.

A Doubly Stochastic Change Point Detection Algorithm for Noisy Biological Signals.

PubMed

Gold, Nathan; Frasch, Martin G; Herry, Christophe L; Richardson, Bryan S; Wang, Xiaogang

2017-01-01

Experimentally and clinically collected time series data are often contaminated with significant confounding noise, creating short, noisy time series. This noise, due to natural variability and measurement error, poses a challenge to conventional change point detection methods. We propose a novel and robust statistical method for change point detection for noisy biological time sequences. Our method is a significant improvement over traditional change point detection methods, which only examine a potential anomaly at a single time point. In contrast, our method considers all suspected anomaly points and considers the joint probability distribution of the number of change points and the elapsed time between two consecutive anomalies. We validate our method with three simulated time series, a widely accepted benchmark data set, two geological time series, a data set of ECG recordings, and a physiological data set of heart rate variability measurements of fetal sheep model of human labor, comparing it to three existing methods. Our method demonstrates significantly improved performance over the existing point-wise detection methods.

Predicting species distributions from checklist data using site-occupancy models

USGS Publications Warehouse

Kery, M.; Gardner, B.; Monnerat, C.

2010-01-01

Aim: (1) To increase awareness of the challenges induced by imperfect detection, which is a fundamental issue in species distribution modelling; (2) to emphasize the value of replicate observations for species distribution modelling; and (3) to show how 'cheap' checklist data in faunal/floral databases may be used for the rigorous modelling of distributions by site-occupancy models. Location: Switzerland. Methods: We used checklist data collected by volunteers during 1999 and 2000 to analyse the distribution of the blue hawker, Aeshna cyanea (Odonata, Aeshnidae), a common dragonfly in Switzerland. We used data from repeated visits to 1-ha pixels to derive 'detection histories' and apply site-occupancy models to estimate the 'true' species distribution, i.e. corrected for imperfect detection. We modelled blue hawker distribution as a function of elevation and year and its detection probability of elevation, year and season. Results: The best model contained cubic polynomial elevation effects for distribution and quadratic effects of elevation and season for detectability. We compared the site-occupancy model with a conventional distribution model based on a generalized linear model, which assumes perfect detectability (p = 1). The conventional distribution map looked very different from the distribution map obtained using site-occupancy models that accounted for the imperfect detection. The conventional model underestimated the species distribution by 60%, and the slope parameters of the occurrence-elevation relationship were also underestimated when assuming p = 1. Elevation was not only an important predictor of blue hawker occurrence, but also of the detection probability, with a bell-shaped relationship. Furthermore, detectability increased over the season. The average detection probability was estimated at only 0.19 per survey. Main conclusions: Conventional species distribution models do not model species distributions per se but rather the apparent distribution, i.e. an unknown proportion of species distributions. That unknown proportion is equivalent to detectability. Imperfect detection in conventional species distribution models yields underestimates of the extent of distributions and covariate effects that are biased towards zero. In addition, patterns in detectability will erroneously be ascribed to species distributions. In contrast, site-occupancy models applied to replicated detection/non-detection data offer a powerful framework for making inferences about species distributions corrected for imperfect detection. The use of 'cheap' checklist data greatly enhances the scope of applications of this useful class of models. ?? 2010 Blackwell Publishing Ltd.

Immunosensors using a quartz crystal microbalance

NASA Astrophysics Data System (ADS)

Kurosawa, Shigeru; Aizawa, Hidenobu; Tozuka, Mitsuhiro; Nakamura, Miki; Park, Jong-Won

2003-11-01

Better analytical technology has been demanded for accurate and rapid determination of trace amounts of chemical compounds, such as marker proteins for disease or endocrine disrupters like dioxin, which might be contained in blood, food and the environment. The study of immunosensors using a quartz crystal microbalance (QCM) has recently focused on conventional detection methods for the determination of chemical compounds together with the development of reagents and processes. This paper introduces the principle of the detection method of QCM immunosensors developed at AIST and its application to the detection of trace amounts of chemical compounds.

Gastric cancer screening of a high-risk population in Japan using serum pepsinogen and barium digital radiography.

PubMed

Ohata, Hiroshi; Oka, Masashi; Yanaoka, Kimihiko; Shimizu, Yasuhito; Mukoubayashi, Chizu; Mugitani, Kouichi; Iwane, Masataka; Nakamura, Hideya; Tamai, Hideyuki; Arii, Kenji; Nakata, Hiroya; Yoshimura, Noriko; Takeshita, Tetsuya; Miki, Kazumasa; Mohara, Osamu; Ichinose, Masao

2005-10-01

With the aim of developing more efficient gastric cancer screening programs for use in Japan, we studied a new screening program that combines serum pepsinogen (PG) testing and barium digital radiography (DR). A total of 17 647 middle-aged male subjects underwent workplace screening over a 7-year period using a combination of PG testing and DR. This program's effectiveness, as well as other characteristics of the program, was analyzed. Forty-nine cases of gastric cancer were detected (comprising 88% early cancer cases). The detection rate was 0.28%, and the positive predictive value was 0.85%. The PG test detected 63.3% of cases, DR detected 69.4% of cases, and both tests were positive in 32.7% of cancer cases. The two methods were almost equally effective, and were considerably more effective than conventional screening using photofluorography. Each screening method detected a distinct gastric cancer subgroup; the PG test efficiently detected asymptomatic small early cancer with intestinal type histology, while DR was efficient at detecting cancers with depressed or ulcerated morphology and diffuse type histology. The cost for the detection of a single cancer was much less than that for conventional screening. In fact, it is possible to further reduce the cost of detecting a single cancer to a cost comparable to that of surgically resecting a single gastric cancer. Thus, it is probable that a highly efficient gastric cancer screening system can be implemented by combining the two screening methods. Such a screening program would be beneficial in a population at high risk for gastric cancer.

Microbial deterioration of vacuum-packaged chilled beef cuts and techniques for microbiota detection and characterization: a review.

PubMed

Hernández-Macedo, Maria Lucila; Barancelli, Giovana Verginia; Contreras-Castillo, Carmen Josefina

2011-01-01

Gas production from microbial deterioration in vacuum-packs of chilled meat leads to pack distension, which is commonly referred as blown pack. This phenomenon is attributed to some psychrophilic and psychrotrophic Clostridium species, as well as Enterobacteria. The ability of these microorganisms to grow at refrigeration temperatures makes the control by the meat industry a challenge. This type of deterioration has been reported in many countries including some plants in the Midwestern and Southeastern regions of Brazil. In addition to causing economic losses, spoilage negatively impacts the commercial product brand, thereby impairing the meat industry. In the case of strict anaerobes species they are difficult to grow and isolate using culture methods in conventional microbiology laboratories. Furthermore, conventional culture methods are sometimes not capable of distinguishing species or genera. DNA-based molecular methods are alternative strategies for detecting viable and non-cultivable microorganisms and strict anaerobic microorganisms that are difficult to cultivate. Here, we review the microorganisms and mechanisms involved in the deterioration of vacuum-packaged chilled meat and address the use of molecular methods for detecting specific strict anaerobic microorganisms and microbial communities in meat samples.

Rapid diagnosis of equine influenza by highly sensitive silver amplification immunochromatography system.

PubMed

Yamanaka, Takashi; Nemoto, Manabu; Bannai, Hiroshi; Tsujimura, Koji; Kondo, Takashi; Matsumura, Tomio; Fu, Tao Qi Huang; Fernandez, Charlene Judith; Gildea, Sarah; Cullinane, Ann

2017-06-16

Equine influenza (EI) is a respiratory disease caused by equine influenza A virus (EIV, H3N8) infection. Rapid diagnosis is essential to limit the disease spread. We previously reported that some rapid antigen detection (RAD) tests are fit for diagnosing EI although their sensitivity is not optimal. Here, we evaluated the performance of the newly developed RAD test using silver amplification immunochromatography (Quick Chaser Auto Flu A, B: QCA) to diagnose EI. The detection limits of QCA for EIVs were five-fold lower than the conventional RAD tests. The duration of virus antigen detection in the infected horses was longer than the conventional RAD tests. We conclude that QCA could be a valuable diagnostic method for EI.

A study on pseudo interface wave technique for CRDM weld defects in nuclear power plants

NASA Astrophysics Data System (ADS)

Lee, Jaesun; Park, Junpil; Cho, Younho; Huh, Hyung; Park, Keun-Bae; Kim, Dong-Ok

2015-03-01

The nuclear power plant inspection is very important for the safety issue. However due to some radiation and geometric problems, the detection of CRDM(Control Rod Drive Mechanism) can be very difficult by using conventional Ultrasonic Testing method. Also the shrink fit boundary condition can also be an obstacle for the inspection in this paper, instead of conventional Ultrasonic Testing, guided wave was used for the detection of some complicated structures. The CRDM nozzle was installed in reactor head with perfect shrink fit condition by using stainless steel. The wave amplitude distribution on the circumferential direction was calculated with various boundary conditions and the experimental result shows a possibility of the defect detection on J-groove weld.

Broad-Range Detection of Microorganisms Directly from Bronchoalveolar Lavage Specimens by PCR/Electrospray Ionization-Mass Spectrometry

PubMed Central

Ullberg, Måns; Lüthje, Petra; Mölling, Paula; Strålin, Kristoffer

2017-01-01

The clinical demand on rapid microbiological diagnostic is constantly increasing. PCR coupled to electrospray ionization-mass spectrometry, PCR/ESI-MS, offers detection and identification of over 750 bacteria and Candida species directly from clinical specimens within 6 hours. In this study, we investigated the clinical performance of the IRIDICA BAC LRT Assay for detection of bacterial pathogens in 121 bronchoalveolar lavage (BAL) samples that were received consecutively at our bacterial laboratory for BAL culture. Commensal or pathogenic microorganisms were detected in 118/121 (98%) BAL samples by PCR/ESI-MS, while in 104/121 (86%) samples by routine culture (P<0.01). Detection of potentially pathogenic microorganisms by PCR/ESI-MS was evaluated in comparison with conventional culture-based or molecular methods. The agreement between positive findings was overall good. Most Staphylococcus aureus-positive PCR/ESI-MS results were confirmed by culture or species-specific PCR (27/33, 82%). The identity of Streptococcus pneumoniae could however be confirmed for only 6/17 (35%) PCR/ESI-MS-positive samples. Non-cultivable and fastidious pathogens, which were not covered by standard culture procedures were readily detected by PCR/ESI-MS, including Legionella pneumophila, Bordetella pertussis, Norcadia species and Mycoplasma pneumoniae. In conclusion, PCR/ESI-MS detected a broad range of potential pathogens with equal or superior sensitivity compared to conventional methods within few hours directly from BAL samples. This novel method might thus provide a relevant tool for diagnostics in critically ill patients. PMID:28085931

Quantitative Mapping of Total Choline in Healthy Human Breast Using Proton Echo Planar Spectroscopic Imaging (PEPSI) at 3 Tesla

PubMed Central

Zhao, Chenguang; Bolan, Patrick J.; Royce, Melanie; Lakkadi, Navneeth; Eberhardt, Steven; Sillerud, Laurel; Lee, Sang-Joon; Posse, Stefan

2012-01-01

Purpose To quantitatively measure tCho levels in healthy breasts using Proton-Echo-Planar-Spectroscopic-Imaging (PEPSI). Material and Methods The 2-dimensional mapping of tCho at 3 Tesla across an entire breast slice using PEPSI and a hybrid spectral quantification method based on LCModel fitting and integration of tCho using the fitted spectrum were developed. This method was validated in 19 healthy females and compared with single voxel spectroscopy (SVS) and with PRESS prelocalized conventional Magnetic Resonance Spectroscopic Imaging (MRSI) using identical voxel size (8 cc) and similar scan times (~7 min). Results A tCho peak with a signal to noise ratio larger than 2 was detected in 10 subjects using both PEPSI and SVS. The average tCho concentration in these subjects was 0.45 ± 0.2 mmol/kg using PEPSI and 0.48±0.3 mmol/kg using SVS. Comparable results were obtained in 2 subjects using conventional MRSI. High lipid content in the spectra of 9 tCho negative subjects was associated with spectral line broadening of more than 26 Hz, which made tCho detection impossible. Conventional MRSI with PRESS prelocalization in glandular tissue in two of these subjects yielded tCho concentrations comparable to PEPSI. Conclusion The detection sensitivity of PEPSI is comparable to SVS and conventional PRESS-MRSI. PEPSI can be potentially used in the evaluation of tCho in breast cancer. A tCho threshold concentration value of ~0.7mmol/kg might be used to differentiate between cancerous and healthy (or benign) breast tissues based on this work and previous studies. PMID:22782667

A comparative assessment of preclinical chemotherapeutic response of tumors using quantitative non-Gaussian diffusion MRI

PubMed Central

Xu, Junzhong; Li, Ke; Smith, R. Adam; Waterton, John C.; Zhao, Ping; Ding, Zhaohua; Does, Mark D.; Manning, H. Charles; Gore, John C.

2016-01-01

Background Diffusion-weighted MRI (DWI) signal attenuation is often not mono-exponential (i.e. non-Gaussian diffusion) with stronger diffusion weighting. Several non-Gaussian diffusion models have been developed and may provide new information or higher sensitivity compared with the conventional apparent diffusion coefficient (ADC) method. However the relative merits of these models to detect tumor therapeutic response is not fully clear. Methods Conventional ADC, and three widely-used non-Gaussian models, (bi-exponential, stretched exponential, and statistical model), were implemented and compared for assessing SW620 human colon cancer xenografts responding to barasertib, an agent known to induce apoptosis via polyploidy. Bayesian Information Criterion (BIC) was used for model selection among all three non-Gaussian models. Results All of tumor volume, histology, conventional ADC, and three non-Gaussian DWI models could show significant differences between control and treatment groups after four days of treatment. However, only the non-Gaussian models detected significant changes after two days of treatment. For any treatment or control group, over 65.7% of tumor voxels indicate the bi-exponential model is strongly or very strongly preferred. Conclusion Non-Gaussian DWI model-derived biomarkers are capable of detecting tumor earlier chemotherapeutic response of tumors compared with conventional ADC and tumor volume. The bi-exponential model provides better fitting compared with statistical and stretched exponential models for the tumor and treatment models used in the current work. PMID:27919785

[Quartz-enhanced photoacoustic spectroscopy trace gas detection system based on the Fabry-Perot demodulation].

PubMed

Lin, Cheng; Zhu, Yong; Wei, Wei; Zhang, Jie; Tian, Li; Xu, Zu-Wen

2013-05-01

An all-optical quartz-enhanced photoacoustic spectroscopy system, based on the F-P demodulation, for trace gas detection in the open environment was proposed. In quartz-enhanced photoacoustic spectroscopy (QEPAS), an optical fiber Fabry-Perot method was used to replace the conventional electronic demodulation method. The photoacoustic signal was obtained by demodulating the variation of the Fabry-Perot cavity between the quartz tuning fork side and the fiber face. An experimental system was setup. The experiment for detection of water vapour in the open environment was carried on. A normalized noise equivalent absorption coefficient of 2.80 x 10(-7) cm(-1) x W x Hz(-1/2) was achieved. The result demonstrated that the sensitivity of the all-optical quartz-enhanced photoacoustic spectroscopy system is about 2.6 times higher than that of the conventional QEPAS system. The all-optical quartz-enhanced photoacoustic spectroscopy system is immune to electromagnetic interference, safe in flammable and explosive gas detection, suitable for high temperature and high humidity environments and realizable for long distance, multi-point and network sensing.

Application of photogrammetry for analysis of occlusal contacts.

PubMed

Shigeta, Yuko; Hirabayashi, Rio; Ikawa, Tomoko; Kihara, Takuya; Ando, Eriko; Hirai, Shinya; Fukushima, Shunji; Ogawa, Takumi

2013-04-01

The conventional 2D-analysis methods for occlusal contacts provided limited information on tooth morphology. This present study aims to detect 3D positional information of occlusal contacts from 2D-photos via photogrammetry. We propose an image processing solution for analysis of occlusal contacts and facets via the black silicone method and a photogrammetric technique. The occlusal facets were reconstructed from a 2D-photograph data-set of inter-occlusal records into a 3D image via photogrammetry. The configuration of the occlusal surface was reproduced with polygons. In addition, the textures of the occlusal contacts were mapped to each polygon. DIFFERENCE FROM CONVENTIONAL METHODS: Constructing occlusal facets with 3D polygons from 2D-photos with photogrammetry was a defining characteristic of this image processing technique. It allowed us to better observe findings of the black silicone method. Compared with conventional 3D analysis using a 3D scanner, our 3D models did not reproduce the detail of the anatomical configuration. However, by merging the findings of the inter-occlusal record, the deformation of mandible and the displacement of periodontal ligaments under occlusal force were reflected in our model. EFFECT OR PERFORMANCE: Through the use of polygons in the conversion of 2D images to 3D images, we were able to define the relation between the location and direction of the occlusal contacts and facets, which was difficult to detect via conventional methods. Through our method of making a 3D polygon model, the findings of inter-occlusal records which reflected the jaw/teeth behavior under occlusal force could be observed 3-dimensionally. Copyright © 2012 Japan Prosthodontic Society. Published by Elsevier Ltd. All rights reserved.

Rapid methods for the detection of foodborne bacterial pathogens: principles, applications, advantages and limitations

PubMed Central

Law, Jodi Woan-Fei; Ab Mutalib, Nurul-Syakima; Chan, Kok-Gan; Lee, Learn-Han

2015-01-01

The incidence of foodborne diseases has increased over the years and resulted in major public health problem globally. Foodborne pathogens can be found in various foods and it is important to detect foodborne pathogens to provide safe food supply and to prevent foodborne diseases. The conventional methods used to detect foodborne pathogen are time consuming and laborious. Hence, a variety of methods have been developed for rapid detection of foodborne pathogens as it is required in many food analyses. Rapid detection methods can be categorized into nucleic acid-based, biosensor-based and immunological-based methods. This review emphasizes on the principles and application of recent rapid methods for the detection of foodborne bacterial pathogens. Detection methods included are simple polymerase chain reaction (PCR), multiplex PCR, real-time PCR, nucleic acid sequence-based amplification (NASBA), loop-mediated isothermal amplification (LAMP) and oligonucleotide DNA microarray which classified as nucleic acid-based methods; optical, electrochemical and mass-based biosensors which classified as biosensor-based methods; enzyme-linked immunosorbent assay (ELISA) and lateral flow immunoassay which classified as immunological-based methods. In general, rapid detection methods are generally time-efficient, sensitive, specific and labor-saving. The developments of rapid detection methods are vital in prevention and treatment of foodborne diseases. PMID:25628612

Validate or falsify: Lessons learned from a microscopy method claimed to be useful for detecting Borrelia and Babesia organisms in human blood.

PubMed

Aase, Audun; Hajdusek, Ondrej; Øines, Øivind; Quarsten, Hanne; Wilhelmsson, Peter; Herstad, Tove K; Kjelland, Vivian; Sima, Radek; Jalovecka, Marie; Lindgren, Per-Eric; Aaberge, Ingeborg S

2016-01-01

A modified microscopy protocol (the LM-method) was used to demonstrate what was interpreted as Borrelia spirochetes and later also Babesia sp., in peripheral blood from patients. The method gained much publicity, but was not validated prior to publication, which became the purpose of this study using appropriate scientific methodology, including a control group. Blood from 21 patients previously interpreted as positive for Borrelia and/or Babesia infection by the LM-method and 41 healthy controls without known history of tick bite were collected, blinded and analysed for these pathogens by microscopy in two laboratories by the LM-method and conventional method, respectively, by PCR methods in five laboratories and by serology in one laboratory. Microscopy by the LM-method identified structures claimed to be Borrelia- and/or Babesia in 66% of the blood samples of the patient group and in 85% in the healthy control group. Microscopy by the conventional method for Babesia only did not identify Babesia in any samples. PCR analysis detected Borrelia DNA in one sample of the patient group and in eight samples of the control group; whereas Babesia DNA was not detected in any of the blood samples using molecular methods. The structures interpreted as Borrelia and Babesia by the LM-method could not be verified by PCR. The method was, thus, falsified. This study underlines the importance of doing proper test validation before new or modified assays are introduced.

Detection of Fusarium verticillioides by PCR-ELISA based on FUM21 gene.

PubMed

Omori, Aline Myuki; Ono, Elisabete Yurie Sataque; Bordini, Jaqueline Gozzi; Hirozawa, Melissa Tiemi; Fungaro, Maria Helena Pelegrinelli; Ono, Mario Augusto

2018-08-01

Fusarium verticillioides is a primary corn pathogen and fumonisin producer which is associated with toxic effects in humans and animals. The traditional methods for detection of fungal contamination based on morphological characteristics are time-consuming and show low sensitivity and specificity. Therefore, the objective of this study was to develop a PCR-ELISA based on the FUM21 gene for F. verticillioides detection. The DNA of the F. verticillioides, Fusarium sp., Aspergillus sp. and Penicillium sp. isolates was analyzed by conventional PCR and PCR-ELISA to determine the specificity. The PCR-ELISA was specific to F. verticillioides isolates, showed a 2.5 pg detection limit and was 100-fold more sensitive than conventional PCR. In corn samples inoculated with F. verticillioides conidia, the detection limit of the PCR-ELISA was 1 × 10 4 conidia/g and was also 100-fold more sensitive than conventional PCR. Naturally contaminated corn samples were analyzed by PCR-ELISA based on the FUM21 gene and PCR-ELISA absorbance values correlated positively (p < 0.05) with Fusarium sp. counts (CFU/g). These results suggest that the PCR-ELISA developed in this study can be useful for F. verticillioides detection in corn samples. Copyright © 2018 Elsevier Ltd. All rights reserved.

Bad data detection in two stage estimation using phasor measurements

NASA Astrophysics Data System (ADS)

Tarali, Aditya

The ability of the Phasor Measurement Unit (PMU) to directly measure the system state, has led to steady increase in the use of PMU in the past decade. However, in spite of its high accuracy and the ability to measure the states directly, they cannot completely replace the conventional measurement units due to high cost. Hence it is necessary for the modern estimators to use both conventional and phasor measurements together. This thesis presents an alternative method to incorporate the new PMU measurements into the existing state estimator in a systematic manner such that no major modification is necessary to the existing algorithm. It is also shown that if PMUs are placed appropriately, the phasor measurements can be used to detect and identify the bad data associated with critical measurements by using this model, which cannot be detected by conventional state estimation algorithm. The developed model is tested on IEEE 14, IEEE 30 and IEEE 118 bus under various conditions.

The evolving role of new imaging methods in breast screening.

PubMed

Houssami, Nehmat; Ciatto, Stefano

2011-09-01

The potential to avert breast cancer deaths through screening means that efforts continue to identify methods which may enhance early detection. While the role of most new imaging technologies remains in adjunct screening or in the work-up of mammography-detected abnormalities, some of the new breast imaging tests (such as MRI) have roles in screening groups of women defined by increased cancer risk. This paper highlights the evidence and the current role of new breast imaging technologies in screening, focusing on those that have broader application in population screening, including digital mammography, breast ultrasound in women with dense breasts, and computer-aided detection. It highlights that evidence on new imaging in screening comes mostly from non-randomised studies that have quantified test detection capability as adjunct to mammography, or have compared measures of screening performance for new technologies with that of conventional mammography. Two RCTs have provided high-quality evidence on the equivalence of digital and conventional mammography and on outcomes of screen-reading complemented by CAD. Many of these imaging technologies enhance cancer detection but also increase recall and false positives in screening. Copyright © 2011 Elsevier Inc. All rights reserved.

Establishment and application of cross-priming isothermal amplification coupled with lateral flow dipstick (CPA-LFD) for rapid and specific detection of red-spotted grouper nervous necrosis virus.

PubMed

Su, Zi Dan; Shi, Cheng Yin; Huang, Jie; Shen, Gui Ming; Li, Jin; Wang, Sheng Qiang; Fan, Chao

2015-09-26

Red-spotted grouper nervous necrosis virus (RGNNV) is an important pathogen that causes diseases in many species of fish in marine aquaculture. The larvae and juveniles are more easily infected by RGNNV and the cumulative mortality is as high as 100 % after being infected with RGNNV. This virus imposes a serious threat to aquaculture of grouper fry. This study aimed to establish a simple, accurate and highly sensitive method for rapid detection of RGNNV on the spot. In this study, the primers specifically targeting RGNNV were designed and cross-priming isothermal amplification (CPA) system was established. The product amplified by CPA was detected through visualization with lateral flow dipstick (LFD). Three important parameters, including the amplification temperature, the concentration of dNTPs and the concentration of Mg(2+) for the CPA system, were optimized. The sensitivity and specificity of this method for RGNNV were tested and compared with those of the conventional RT-PCR and real-time quantitative RT-PCR (qRT-PCR). The optimized conditions for the CPA amplification system were determined as follows: the optimal amplification temperature, the optimized concentration of dNTPs and the concentration for Mg(2+) were 69 °C, 1.2 mmol/L and 5 mmol/L, respectively. The lowest limit of detection (LLOD) of this method for RGNNV was 10(1) copies/μL of RNA sample, which was 10 times lower than that of conventional RT-PCR and comparable to that of RT-qPCR. This method was specific for RGNNV in combination with SJNNV and had no cross-reactions with 8 types of virus and bacterial strains tested. This method was successfully applied to detect RGNNV in fish samples. This study established a CPA-LFD method for detection of RGNNV. This method is simple and rapid with high sensitivity and good specificity and can be widely applied for rapid detection of this virus on the spot.

A Novel Technique to Detect Code for SAC-OCDMA System

NASA Astrophysics Data System (ADS)

Bharti, Manisha; Kumar, Manoj; Sharma, Ajay K.

2018-04-01

The main task of optical code division multiple access (OCDMA) system is the detection of code used by a user in presence of multiple access interference (MAI). In this paper, new method of detection known as XOR subtraction detection for spectral amplitude coding OCDMA (SAC-OCDMA) based on double weight codes has been proposed and presented. As MAI is the main source of performance deterioration in OCDMA system, therefore, SAC technique is used in this paper to eliminate the effect of MAI up to a large extent. A comparative analysis is then made between the proposed scheme and other conventional detection schemes used like complimentary subtraction detection, AND subtraction detection and NAND subtraction detection. The system performance is characterized by Q-factor, BER and received optical power (ROP) with respect to input laser power and fiber length. The theoretical and simulation investigations reveal that the proposed detection technique provides better quality factor, security and received power in comparison to other conventional techniques. The wide opening of eye in case of proposed technique also proves its robustness.

Issues in identifying germ tube positive yeasts by conventional methods.

PubMed

Yazdanpanah, Atta; Khaithir, Tzar Mohd Nizam

2014-01-01

Candida speciation is vital for epidemiology and management of candidiasis. Nonmolecular conventional methods often fail to identify closely related germ tube positive yeasts from clinical specimens. The present study was conducted to identify these yeasts and to highlight issues in conventional versus molecular methods of identification. A total of 98 germ tube positive yeasts from high vaginal swabs were studied over a 12-month period. Isolates were examined with various methods including growth at 42 °C and 45 °C on Sabouraud dextrose agar (SDA), color development on CHROMagar Candida medium, chlamydospore production on corn meal agar at 25 °C, carbohydrate assimilation using ID 32C system, and polymerase chain reaction using a single pair of primers targeting the hyphal wall protein 1 (Hwp1) gene. Of all the isolates studied, 97 were molecularly confirmed as C. albicans and one isolate was identified as C. dubliniensis. No C. africana was detected in this study. The molecular method used in our study was an accurate and useful tool for discriminating C. albicans, C. dubliniensis, and C. africana. The conventional methods, however, were less accurate and riddled with many issues that will be discussed in further details. © 2013 Wiley Periodicals, Inc.

Detection of HIV-1 p24 Gag in plasma by a nanoparticle-based bio-barcode-amplification method.

PubMed

Kim, Eun-Young; Stanton, Jennifer; Korber, Bette T M; Krebs, Kendall; Bogdan, Derek; Kunstman, Kevin; Wu, Samuel; Phair, John P; Mirkin, Chad A; Wolinsky, Steven M

2008-06-01

Detection of HIV-1 in patients is limited by the sensitivity and selectivity of available tests. The nanotechnology-based bio-barcode-amplification method offers an innovative approach to detect specific HIV-1 antigens from diverse HIV-1 subtypes. We evaluated the efficacy of this protein-detection method in detecting HIV-1 in men enrolled in the Chicago component of the Multicenter AIDS Cohort Study (MACS). The method relies on magnetic microparticles with antibodies that specifically bind the HIV-1 p24 Gag protein and nanoparticles that are encoded with DNA and antibodies that can sandwich the target protein captured by the microparticle-bound antibodies. The aggregate sandwich structures are magnetically separated from solution, and treated to remove the conjugated barcode DNA. The DNA barcodes (hundreds per target) were identified by a nanoparticle-based detection method that does not rely on PCR. Of 112 plasma samples from HIV-1-infected subjects, 111 were positive for HIV-1 p24 Gag protein (range: 0.11-71.5 ng/ml of plasma) by the bio-barcode-amplification method. HIV-1 p24 Gag protein was detected in only 23 out of 112 men by the conventional ELISA. A total of 34 uninfected subjects were negative by both tests. Thus, the specificity of the bio-barcode-amplification method was 100% and the sensitivity 99%. The bio-barcode-amplification method detected HIV-1 p24 Gag protein in plasma from all study subjects with less than 200 CD4(+) T cells/microl of plasma (100%) and 19 out of 20 (95%) HIV-1-infected men who had less than 50 copies/ml of plasma of HIV-1 RNA. In a separate group of 60 diverse international isolates, representative of clades A, B, C and D and circulating recombinant forms CRF01_AE and CRF02_AG, the bio-barcode-amplification method identified the presence of virus correctly. The bio-barcode-amplification method was superior to the conventional ELISA assay for the detection of HIV-1 p24 Gag protein in plasma with a breadth of coverage for diverse HIV-1 subtypes. Because the bio-barcode-amplification method does not require enzymatic amplification, this method could be translated into a robust point-of-care test.

Novel HPLC Analysis of Hydrocortisone in Conventional and Controlled-Release Pharmaceutical Preparations.

PubMed

Adi-Dako, Ofosua; Oppong Bekoe, Samuel; Ofori-Kwakye, Kwabena; Appiah, Enoch; Peprah, Paul

2017-01-01

An isocratic sensitive and precise reverse phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the determination and quantification of hydrocortisone in controlled-release and conventional (tablets and injections) pharmaceutical preparations. Chromatographic separation was achieved on an ODS (C18), 5  μ m, 4.6 × 150 mm, with an isocratic elution using a freshly prepared mobile phase of composition methanol : water : acetic acid (60 : 30 : 10, v/v/v) at a flow rate of 1.0 ml/min. The detection of the drug was successfully achieved at a wavelength of 254 nm. The retention time obtained for the drug was 2.26 min. The proposed method produced linear detectable responses in the concentration range of 0.02 to 0.4 mg/ml of hydrocortisone. High recoveries of 98-101% were attained at concentration levels of 80%, 100%, and 120%. The intraday and interday precision (RSD) were 0.19-0.55% and 0.33-0.71%, respectively. A comparison of hydrocortisone analyses data from the developed method and the official USP method showed no significant difference ( p > 0.05) at a 95% confidence interval. The method was successfully applied to the determination and quantification of hydrocortisone in six controlled-release and fifteen conventional release pharmaceutical preparations.

Novel HPLC Analysis of Hydrocortisone in Conventional and Controlled-Release Pharmaceutical Preparations

PubMed Central

Oppong Bekoe, Samuel; Appiah, Enoch; Peprah, Paul

2017-01-01

An isocratic sensitive and precise reverse phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the determination and quantification of hydrocortisone in controlled-release and conventional (tablets and injections) pharmaceutical preparations. Chromatographic separation was achieved on an ODS (C18), 5 μm, 4.6 × 150 mm, with an isocratic elution using a freshly prepared mobile phase of composition methanol : water : acetic acid (60 : 30 : 10, v/v/v) at a flow rate of 1.0 ml/min. The detection of the drug was successfully achieved at a wavelength of 254 nm. The retention time obtained for the drug was 2.26 min. The proposed method produced linear detectable responses in the concentration range of 0.02 to 0.4 mg/ml of hydrocortisone. High recoveries of 98–101% were attained at concentration levels of 80%, 100%, and 120%. The intraday and interday precision (RSD) were 0.19–0.55% and 0.33–0.71%, respectively. A comparison of hydrocortisone analyses data from the developed method and the official USP method showed no significant difference (p > 0.05) at a 95% confidence interval. The method was successfully applied to the determination and quantification of hydrocortisone in six controlled-release and fifteen conventional release pharmaceutical preparations. PMID:28660092

A novel kit for rapid detection of Vibrio cholerae O1.

PubMed

Hasan, J A; Huq, A; Tamplin, M L; Siebeling, R J; Colwell, R R

1994-01-01

We report on the development and testing of a novel, rapid, colorimetric immunodiagnostic kit, Cholera SMART, for direct detection of the presence of Vibrio cholerae O1 in clinical specimens. Unlike conventional culture methods requiring several days to complete, the Cholera SMART kit can be used directly in the field by untrained or minimally skilled personnel to detect V. cholerae O1 in less than 15 min, without cumbersome laboratory equipment. A total of 120 clinical and environmental bacterial strains, including both O1 and non-O1 serotypes of V. cholerae isolated from samples collected from a variety of geographical regions, were tested, and positive reactions were observed only with V. cholerae O1. Also, results of a field trial in Bangladesh, employing Cholera SMART, showed 100% specificity and 96% sensitivity compared with conventional culture methods. Another field trial, in Mexico, showed that Cholera SMART was 100% in agreement with a recently described coagglutination test when 108 stool specimens were tested.

Light Weight MP3 Watermarking Method for Mobile Terminals

NASA Astrophysics Data System (ADS)

Takagi, Koichi; Sakazawa, Shigeyuki; Takishima, Yasuhiro

This paper proposes a novel MP3 watermarking method which is applicable to a mobile terminal with limited computational resources. Considering that in most cases the embedded information is copyright information or metadata, which should be extracted before playing back audio contents, the watermark detection process should be executed at high speed. However, when conventional methods are used with a mobile terminal, it takes a considerable amount of time to detect a digital watermark. This paper focuses on scalefactor manipulation to enable high speed watermark embedding/detection for MP3 audio and also proposes the manipulation method which minimizes audio quality degradation adaptively. Evaluation tests showed that the proposed method is capable of embedding 3 bits/frame information without degrading audio quality and detecting it at very high speed. Finally, this paper describes application examples for authentication with a digital signature.

Road Lane Detection by Discriminating Dashed and Solid Road Lanes Using a Visible Light Camera Sensor.

PubMed

Hoang, Toan Minh; Hong, Hyung Gil; Vokhidov, Husan; Park, Kang Ryoung

2016-08-18

With the increasing need for road lane detection used in lane departure warning systems and autonomous vehicles, many studies have been conducted to turn road lane detection into a virtual assistant to improve driving safety and reduce car accidents. Most of the previous research approaches detect the central line of a road lane and not the accurate left and right boundaries of the lane. In addition, they do not discriminate between dashed and solid lanes when detecting the road lanes. However, this discrimination is necessary for the safety of autonomous vehicles and the safety of vehicles driven by human drivers. To overcome these problems, we propose a method for road lane detection that distinguishes between dashed and solid lanes. Experimental results with the Caltech open database showed that our method outperforms conventional methods.

Road Lane Detection by Discriminating Dashed and Solid Road Lanes Using a Visible Light Camera Sensor

PubMed Central

Hoang, Toan Minh; Hong, Hyung Gil; Vokhidov, Husan; Park, Kang Ryoung

2016-01-01

With the increasing need for road lane detection used in lane departure warning systems and autonomous vehicles, many studies have been conducted to turn road lane detection into a virtual assistant to improve driving safety and reduce car accidents. Most of the previous research approaches detect the central line of a road lane and not the accurate left and right boundaries of the lane. In addition, they do not discriminate between dashed and solid lanes when detecting the road lanes. However, this discrimination is necessary for the safety of autonomous vehicles and the safety of vehicles driven by human drivers. To overcome these problems, we propose a method for road lane detection that distinguishes between dashed and solid lanes. Experimental results with the Caltech open database showed that our method outperforms conventional methods. PMID:27548176

Laser fluorometric analysis of plants for uranium exploration

USGS Publications Warehouse

Harms, T.F.; Ward, F.N.; Erdman, J.A.

1981-01-01

A preliminary test of biogeochemical exploration for locating uranium occurrences in the Marfa Basin, Texas, was conducted in 1978. Only 6 of 74 plant samples (mostly catclaw mimosa, Mimosa biuncifera) contained uranium in amounts above the detection limit (0.4 ppm in the ash) of the conventional fluorometric method. The samples were then analyzed using a Scintrex UA-3 uranium analyzer* * Use of trade names in this paper is for descriptive purposes only and does not constitute endorsement by the U.S. Geological Survey. - an instrument designed for direct analysis of uranium in water, and which can be conveniently used in a mobile field laboratory. The detection limit for uranium in plant ash (0.05 ppm) by this method is almost an order of magnitude lower than with the fluorometric conventional method. Only 1 of the 74 samples contained uranium below the detection limit of the new method. Accuracy and precision were determined to be satisfactory. Samples of plants growing on mineralized soils and nonmineralized soils show a 15-fold difference in uranium content; whereas the soils themselves (analyzed by delayed neutron activation analysis) show only a 4-fold difference. The method involves acid digestion of ashed tissue, extraction of uranium into ethyl acetate, destruction of the ethyl acetate, dissolution of the residue in 0.005% nitric acid, and measurement. ?? 1981.

RECOVERY ACT: MULTIMODAL IMAGING FOR SOLAR CELL MICROCRACK DETECTION

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

Janice Hudgings; Lawrence Domash

2012-02-08

Undetected microcracks in solar cells are a principal cause of failure in service due to subsequent weather exposure, mechanical flexing or diurnal temperature cycles. Existing methods have not been able to detect cracks early enough in the production cycle to prevent inadvertent shipment to customers. This program, sponsored under the DOE Photovoltaic Supply Chain and Cross-Cutting Technologies program, studied the feasibility of quantifying surface micro-discontinuities by use of a novel technique, thermoreflectance imaging, to detect surface temperature gradients with very high spatial resolution, in combination with a suite of conventional imaging methods such as electroluminescence. The project carried out laboratorymore » tests together with computational image analyses using sample solar cells with known defects supplied by industry sources or DOE National Labs. Quantitative comparisons between the effectiveness of the new technique and conventional methods were determined in terms of the smallest detectable crack. Also the robustness of the new technique for reliable microcrack detection was determined at various stages of processing such as before and after antireflectance treatments. An overall assessment is that the new technique compares favorably with existing methods such as lock-in thermography or ultrasonics. The project was 100% completed in Sept, 2010. A detailed report of key findings from this program was published as: Q.Zhou, X.Hu, K.Al-Hemyari, K.McCarthy, L.Domash and J.Hudgings, High spatial resolution characterization of silicon solar cells using thermoreflectance imaging, J. Appl. Phys, 110, 053108 (2011).« less

Automatic detection method for mura defects on display film surface using modified Weber's law

NASA Astrophysics Data System (ADS)

Kim, Myung-Muk; Lee, Seung-Ho

2014-07-01

We propose a method that automatically detects mura defects on display film surfaces using a modified version of Weber's law. The proposed method detects mura defects regardless of their properties and shapes by identifying regions perceived by human vision as mura using the brightness of pixel and image distribution ratio of mura in an image histogram. The proposed detection method comprises five stages. In the first stage, the display film surface image is acquired and a gray-level shift performed. In the second and third stages, the image histogram is acquired and analyzed, respectively. In the fourth stage, the mura range is acquired. This is followed by postprocessing in the fifth stage. Evaluations of the proposed method conducted using 200 display film mura image samples indicate a maximum detection rate of ˜95.5%. Further, the results of application of the Semu index for luminance mura in flat panel display (FPD) image quality inspection indicate that the proposed method is more reliable than a popular conventional method.

Shadow Detection Based on Regions of Light Sources for Object Extraction in Nighttime Video

PubMed Central

Lee, Gil-beom; Lee, Myeong-jin; Lee, Woo-Kyung; Park, Joo-heon; Kim, Tae-Hwan

2017-01-01

Intelligent video surveillance systems detect pre-configured surveillance events through background modeling, foreground and object extraction, object tracking, and event detection. Shadow regions inside video frames sometimes appear as foreground objects, interfere with ensuing processes, and finally degrade the event detection performance of the systems. Conventional studies have mostly used intensity, color, texture, and geometric information to perform shadow detection in daytime video, but these methods lack the capability of removing shadows in nighttime video. In this paper, a novel shadow detection algorithm for nighttime video is proposed; this algorithm partitions each foreground object based on the object’s vertical histogram and screens out shadow objects by validating their orientations heading toward regions of light sources. From the experimental results, it can be seen that the proposed algorithm shows more than 93.8% shadow removal and 89.9% object extraction rates for nighttime video sequences, and the algorithm outperforms conventional shadow removal algorithms designed for daytime videos. PMID:28327515

Ictal SPECT using an attachable automated injector: clinical usefulness in the prediction of ictal onset zone.

PubMed

Lee, Jung-Ju; Lee, Sang Kun; Choi, Jang Wuk; Kim, Dong-Wook; Park, Kyung Il; Kim, Bom Sahn; Kang, Hyejin; Lee, Dong Soo; Lee, Seo-Young; Kim, Sung Hun; Chung, Chun Kee; Nam, Hyeon Woo; Kim, Kwang Ki

2009-12-01

Ictal single-photon emission computed tomography (SPECT) is a valuable method for localizing the ictal onset zone in the presurgical evaluation of patients with intractable epilepsy. Conventional methods used to localize the ictal onset zone have problems with time lag from seizure onset to injection. To evaluate the clinical usefulness of a method that we developed, which involves an attachable automated injector (AAI), in reducing time lag and improving the ability to localize the zone of seizure onset. Patients admitted to the epilepsy monitoring unit (EMU) between January 1, 2003, and June 30, 2008, were included. The definition of ictal onset zone was made by comprehensive review of medical records, magnetic resonance imaging (MRI), data from video electroencephalography (EEG) monitoring, and invasive EEG monitoring if available. We comprehensively evaluated the time lag to injection and the image patterns of ictal SPECT using traditional visual analysis, statistical parametric mapping-assisted, and subtraction ictal SPECT coregistered to an MRI-assisted means of analysis. Image patterns were classified as localizing, lateralizing, and nonlateralizing. The whole number of patients was 99: 48 in the conventional group and 51 in the AAI group. The mean (SD) delay time to injection from seizure onset was 12.4+/-12.0 s in the group injected by our AAI method and 40.4+/-26.3 s in the group injected by the conventional method (P=0.000). The mean delay time to injection from seizure detection was 3.2+/-2.5 s in the group injected by the AAI method and 21.4+/-9.7 s in the group injected by the conventional method (P=0.000). The AAI method was superior to the conventional method in localizing the area of seizure onset (36 out of 51 with AAI method vs. 21 out of 48 with conventional method, P=0.009), especially in non-temporal lobe epilepsy (non-TLE) patients (17 out of 27 with AAI method vs. 3 out of 13 with conventional method, P=0.041), and in lateralizing the seizure onset hemisphere (47 out of 51 with AAI method vs. 33 out of 48 with conventional method, P=0.004). The AAI method was superior to the conventional method in reducing the time lag of tracer injection and in localizing and lateralizing the ictal onset zone, especially in patients with non-TLE.

Performance and cost analysis of Siriraj liquid-based cytology: a direct-to-vial study.

PubMed

Laiwejpithaya, Somsak; Benjapibal, Mongkol; Laiwejpithaya, Sujera; Wongtiraporn, Weerasak; Sangkarat, Suthi; Rattanachaiyanont, Manee

2009-12-01

To compare the cytological diagnoses, specimen adequacy, and cost of the Siriraj liquid-based cytology (LBC) with those of the conventional smear technique. An observational study with historical comparison was conducted in a tertiary university hospital. Cytological reports of 23,676 Siriraj-LBC specimens obtained in 2006 were compared with those of 25,510 conventional smears obtained in 2004. Overall prevalence of abnormal cervical cytology detected by conventional smear was 1.76% and by Siriraj-LBC was 3.70%. Compared with the conventional method, the Siriraj-LBC yielded a significantly higher overall detection rate of abnormal cervical cytology, with a 110.23% increase in the detection rate (P<0.001), mainly due to the increase in diagnosis of squamous intraepithelial lesions (SIL), both low and high grade, together with atypical squamous cells of undetermined significance, "atypical squamous cells cannot exclude HSIL", and malignancies, but not atypical glandular cells. The Siriraj-LBC had a smaller proportion of unsatisfactory slides (4.94% vs. 18.60%, P<0.001) and a higher negative predictive value (96.33% vs. 92.74%, P=0.001), but no difference in positive predictive value (83.03% vs. 86.83%, P=0.285). The cost of Siriraj-LBC was approximately 67% higher than that of the conventional cytology used in Siriraj Hospital and 50-70% lower than that of the commercially available LBC techniques in Thailand. The Siriraj-LBC increases the detection rate of abnormal cytology, improves specimen adequacy, and enhances the negative predictive value without compromising the positive predictive value. For centers where conventional Pap smear does not perform well, the introduction of a low cost Siriraj-LBC might help to improve performance and it may be an economical alternative to the commercially available liquid-based cytology.

Data based abnormality detection

NASA Astrophysics Data System (ADS)

Purwar, Yashasvi

Data based abnormality detection is a growing research field focussed on extracting information from feature rich data. They are considered to be non-intrusive and non-destructive in nature which gives them a clear advantage over conventional methods. In this study, we explore different streams of data based anomalies detection. We propose extension and revisions to existing valve stiction detection algorithm supported with industrial case study. We also explored the area of image analysis and proposed a complete solution for Malaria diagnosis. The proposed method is tested over images provided by pathology laboratory at Alberta Health Service. We also address the robustness and practicality of the solution proposed.

Pick- and waveform-based techniques for real-time detection of induced seismicity

NASA Astrophysics Data System (ADS)

Grigoli, Francesco; Scarabello, Luca; Böse, Maren; Weber, Bernd; Wiemer, Stefan; Clinton, John F.

2018-05-01

The monitoring of induced seismicity is a common operation in many industrial activities, such as conventional and non-conventional hydrocarbon production or mining and geothermal energy exploitation, to cite a few. During such operations, we generally collect very large and strongly noise-contaminated data sets that require robust and automated analysis procedures. Induced seismicity data sets are often characterized by sequences of multiple events with short interevent times or overlapping events; in these cases, pick-based location methods may struggle to correctly assign picks to phases and events, and errors can lead to missed detections and/or reduced location resolution and incorrect magnitudes, which can have significant consequences if real-time seismicity information are used for risk assessment frameworks. To overcome these issues, different waveform-based methods for the detection and location of microseismicity have been proposed. The main advantages of waveform-based methods is that they appear to perform better and can simultaneously detect and locate seismic events providing high-quality locations in a single step, while the main disadvantage is that they are computationally expensive. Although these methods have been applied to different induced seismicity data sets, an extensive comparison with sophisticated pick-based detection methods is still missing. In this work, we introduce our improved waveform-based detector and we compare its performance with two pick-based detectors implemented within the SeiscomP3 software suite. We test the performance of these three approaches with both synthetic and real data sets related to the induced seismicity sequence at the deep geothermal project in the vicinity of the city of St. Gallen, Switzerland.

A Sparsity-based Framework for Resolution Enhancement in Optical Fault Analysis of Integrated Circuits

DTIC Science & Technology

2015-01-01

for IC fault detection . This section provides background information on inversion methods. Conventional inversion techniques and their shortcomings are...physical techniques, electron beam imaging/analysis, ion beam techniques, scanning probe techniques. Electrical tests are used to detect faults in 13 an...hand, there is also the second harmonic technique through which duty cycle degradation faults are detected by collecting the magnitude and the phase of

Detection of soft tissue densities from digital breast tomosynthesis: comparison of conventional and deep learning approaches

NASA Astrophysics Data System (ADS)

Fotin, Sergei V.; Yin, Yin; Haldankar, Hrishikesh; Hoffmeister, Jeffrey W.; Periaswamy, Senthil

2016-03-01

Computer-aided detection (CAD) has been used in screening mammography for many years and is likely to be utilized for digital breast tomosynthesis (DBT). Higher detection performance is desirable as it may have an impact on radiologist's decisions and clinical outcomes. Recently the algorithms based on deep convolutional architectures have been shown to achieve state of the art performance in object classification and detection. Similarly, we trained a deep convolutional neural network directly on patches sampled from two-dimensional mammography and reconstructed DBT volumes and compared its performance to a conventional CAD algorithm that is based on computation and classification of hand-engineered features. The detection performance was evaluated on the independent test set of 344 DBT reconstructions (GE SenoClaire 3D, iterative reconstruction algorithm) containing 328 suspicious and 115 malignant soft tissue densities including masses and architectural distortions. Detection sensitivity was measured on a region of interest (ROI) basis at the rate of five detection marks per volume. Moving from conventional to deep learning approach resulted in increase of ROI sensitivity from 0:832 +/- 0:040 to 0:893 +/- 0:033 for suspicious ROIs; and from 0:852 +/- 0:065 to 0:930 +/- 0:046 for malignant ROIs. These results indicate the high utility of deep feature learning in the analysis of DBT data and high potential of the method for broader medical image analysis tasks.

Denoising embolic Doppler ultrasound signals using Dual Tree Complex Discrete Wavelet Transform.

PubMed

Serbes, Gorkem; Aydin, Nizamettin

2010-01-01

Early and accurate detection of asymptomatic emboli is important for monitoring of preventive therapy in stroke-prone patients. One of the problems in detection of emboli is the identification of an embolic signal caused by very small emboli. The amplitude of the embolic signal may be so small that advanced processing methods are required to distinguish these signals from Doppler signals arising from red blood cells. In this study instead of conventional discrete wavelet transform, the Dual Tree Complex Discrete Wavelet Transform was used for denoising embolic signals. Performances of both approaches were compared. Unlike the conventional discrete wavelet transform discrete complex wavelet transform is a shift invariant transform with limited redundancy. Results demonstrate that the Dual Tree Complex Discrete Wavelet Transform based denoising outperforms conventional discrete wavelet denoising. Approximately 8 dB improvement is obtained by using the Dual Tree Complex Discrete Wavelet Transform compared to the improvement provided by the conventional Discrete Wavelet Transform (less than 5 dB).

Improved detection of Burkholderia pseudomallei from non-blood clinical specimens using enrichment culture and PCR: narrowing diagnostic gap in resource-constrained settings.

PubMed

Tellapragada, Chaitanya; Shaw, Tushar; D'Souza, Annet; Eshwara, Vandana Kalwaje; Mukhopadhyay, Chiranjay

2017-07-01

To evaluate the diagnostic utility of enrichment culture and PCR for improved case detection rates of non-bacteraemic form of melioidosis in limited resource settings. Clinical specimens (n = 525) obtained from patients presenting at a tertiary care hospital of South India with clinical symptoms suggestive of community-acquired pneumonia, lower respiratory tract infections, superficial or internal abscesses, chronic skin ulcers and bone or joint infections were tested for the presence of Burkholderia pseudomallei using conventional culture (CC), enrichment culture (EC) and PCR. Sensitivity, specificity, positive and negative predictive values of CC and PCR were initially deduced using EC as the gold standard method. Further, diagnostic accuracies of all the three methods were analysed using Bayesian latent class modelling (BLCM). Detection rates of B. pseudomallei using CC, EC and PCR were 3.8%, 5.3% and 6%, respectively. Diagnostic sensitivities and specificities of CC and PCR were 71.4, 98.4% and 100 and 99.4%, respectively in comparison with EC as the gold standard test. With Bayesian latent class modelling, EC and PCR demonstrated sensitivities of 98.7 and 99.3%, respectively, while CC showed a sensitivity of 70.3% for detection of B. pseudomallei. An increase of 1.6% (95% CI: 1.08-4.32%) in the case detection rate of melioidosis was observed in the study population when EC and/or PCR were used in adjunct to the conventional culture technique. Our study findings underscore the diagnostic superiority of enrichment culture and/or PCR over conventional microbiological culture for improved case detection of melioidosis from non-blood clinical specimens. © 2017 John Wiley & Sons Ltd.

Coded Excitation Plane Wave Imaging for Shear Wave Motion Detection

PubMed Central

Song, Pengfei; Urban, Matthew W.; Manduca, Armando; Greenleaf, James F.; Chen, Shigao

2015-01-01

Plane wave imaging has greatly advanced the field of shear wave elastography thanks to its ultrafast imaging frame rate and the large field-of-view (FOV). However, plane wave imaging also has decreased penetration due to lack of transmit focusing, which makes it challenging to use plane waves for shear wave detection in deep tissues and in obese patients. This study investigated the feasibility of implementing coded excitation in plane wave imaging for shear wave detection, with the hypothesis that coded ultrasound signals can provide superior detection penetration and shear wave signal-to-noise-ratio (SNR) compared to conventional ultrasound signals. Both phase encoding (Barker code) and frequency encoding (chirp code) methods were studied. A first phantom experiment showed an approximate penetration gain of 2-4 cm for the coded pulses. Two subsequent phantom studies showed that all coded pulses outperformed the conventional short imaging pulse by providing superior sensitivity to small motion and robustness to weak ultrasound signals. Finally, an in vivo liver case study on an obese subject (Body Mass Index = 40) demonstrated the feasibility of using the proposed method for in vivo applications, and showed that all coded pulses could provide higher SNR shear wave signals than the conventional short pulse. These findings indicate that by using coded excitation shear wave detection, one can benefit from the ultrafast imaging frame rate and large FOV provided by plane wave imaging while preserving good penetration and shear wave signal quality, which is essential for obtaining robust shear elasticity measurements of tissue. PMID:26168181

Detecting free-mass common-mode motion induced by incident gravitational waves

NASA Astrophysics Data System (ADS)

Tobar, Michael Edmund; Suzuki, Toshikazu; Kuroda, Kazuaki

1999-05-01

In this paper we show that information on both the differential and common mode free-mass response to a gravitational wave can provide important information on discriminating the direction of the gravitational wave source and between different theories of gravitation. The conventional Michelson interferometer scheme only measures the differential free-mass response. By changing the orientation of the beam splitter, it is possible to configure the detector so it is sensitive to the common-mode of the free-mass motion. The proposed interferometer is an adaptation of the Fox-Smith interferometer. A major limitation to the new scheme is its enhanced sensitivity to laser frequency fluctuations over the conventional, and we propose a method of cancelling these fluctuations. The configuration could be used in parallel to the conventional differential detection scheme with a significant sensitivity and bandwidth.

On-line high-performance liquid chromatography-ultraviolet-nuclear magnetic resonance method of the markers of nerve agents for verification of the Chemical Weapons Convention.

PubMed

Mazumder, Avik; Gupta, Hemendra K; Garg, Prabhat; Jain, Rajeev; Dubey, Devendra K

2009-07-03

This paper details an on-flow liquid chromatography-ultraviolet-nuclear magnetic resonance (LC-UV-NMR) method for the retrospective detection and identification of alkyl alkylphosphonic acids (AAPAs) and alkylphosphonic acids (APAs), the markers of the toxic nerve agents for verification of the Chemical Weapons Convention (CWC). Initially, the LC-UV-NMR parameters were optimized for benzyl derivatives of the APAs and AAPAs. The optimized parameters include stationary phase C(18), mobile phase methanol:water 78:22 (v/v), UV detection at 268nm and (1)H NMR acquisition conditions. The protocol described herein allowed the detection of analytes through acquisition of high quality NMR spectra from the aqueous solution of the APAs and AAPAs with high concentrations of interfering background chemicals which have been removed by preceding sample preparation. The reported standard deviation for the quantification is related to the UV detector which showed relative standard deviations (RSDs) for quantification within +/-1.1%, while lower limit of detection upto 16mug (in mug absolute) for the NMR detector. Finally the developed LC-UV-NMR method was applied to identify the APAs and AAPAs in real water samples, consequent to solid phase extraction and derivatization. The method is fast (total experiment time approximately 2h), sensitive, rugged and efficient.

Inductance and resistance measurement method for vessel detection and coil powering in all-surface inductive heating systems composed of outer squircle coils

NASA Astrophysics Data System (ADS)

Kilic, Veli Tayfun; Unal, Emre; Demir, Hilmi Volkan

2017-05-01

In this work, we investigate a method proposed for vessel detection and coil powering in an all-surface inductive heating system composed of outer squircle coils. Besides conventional circular coils, coils with different shapes such as outer squircle coils are used for and enable efficient all-surface inductive heating. Validity of the method, which relies on measuring inductance and resistance values of a loaded coil at different frequencies, is experimentally demonstrated for a coil with shape different from conventional circular coil. Simple setup was constructed with a small coil to model an all-surface inductive heating system. Inductance and resistance maps were generated by measuring coil's inductance and resistance values at different frequencies loaded by a plate made of different materials and located at various positions. Results show that in an induction hob for various coil geometries it is possible to detect a vessel's presence, to identify its material type and to specify its position on the hob surface by considering inductance and resistance of the coil measured on at least two different frequencies. The studied method is important in terms of enabling safe, efficient and user flexible heating in an all-surface inductive heating system by automatically detecting the vessel's presence and powering on only the coils that are loaded by the vessel with predetermined current levels.

Establishment and Application of a Loop-Mediated Isothermal Amplification Method for Simple, Specific, Sensitive and Rapid Detection of Toxoplasma gondii

PubMed Central

CAO, Lili; CHENG, Ronghua; YAO, Lin; YUAN, Shuxian; YAO, Xinhua

2013-01-01

ABSTRACT The Loop-mediated isothermal amplification (LAMP) method amplifies DNA with high simply, specificity, sensitivity and rapidity. In this study, A LAMP assay with 6 primers targeting a highly conserved region of the GRA1 gene was developed to diagnose Toxoplasma gondii. The reaction time of the LAMP assay was shortened to 30 min after optimizing the reaction system. The LAMP assay was found to be highly specific and stable. The detection limit of the LAMP assay was 10 copies, the same as that of the conventional PCR. We used the LAMP assay to develop a real-time fluorogenic protocol to quantitate T. gondii DNA and generated a log-linear regression plot by plotting the time-to-threshold values against genomic equivalent copies. Furthermore, the LAMP assay was applied to detect T. gondii DNA in 423 blood samples and 380 lymph node samples from 10 pig farms, and positive results were obtained for 7.8% and 8.2% of samples, respectively. The results showed that the LAMP method is slightly more sensitive than conventional PCR (6.1% and 7.6%). Positive samples obtained from 6 pig farms. The LAMP assay established in this study resulted in simple, specific, sensitive and rapid detection of T. gondii DNA and is expected to play an important role in clinical detection of T. gondii. PMID:23965849

Rapid detection of Porcine circovirus 2 by recombinase polymerase amplification.

PubMed

Wang, Jianchang; Wang, Jinfeng; Liu, Libing; Li, Ruiwen; Yuan, Wanzhe

2016-09-01

Porcine circovirus-associated disease, caused primarily by Porcine circovirus 2 (PCV-2), has become endemic in many pig-producing countries and has resulted in significant economic losses to the swine industry worldwide. Tests for PCV-2 infection include PCR, nested PCR, competitive PCR, and real-time PCR (rtPCR). Recombinase polymerase amplification (RPA) has emerged as an isothermal gene amplification technology for the molecular detection of infectious disease agents. RPA is performed at a constant temperature and therefore can be carried out in a water bath. In addition, RPA is completed in ~30 min, much faster than PCR, which usually takes >60 min. We developed a RPA-based method for the detection of PCV-2. The detection limit of RPA was 10(2) copies of PCV-2 genomic DNA. RPA showed the same sensitivity as rtPCR but was 10 times more sensitive than conventional PCR. Successful amplification of PCV-2 DNA, but not other viral templates, demonstrated high specificity of the RPA assay. This method was also validated using clinical samples. The results showed that the RPA assay had a diagnostic agreement rate of 93.7% with conventional PCR and 100% with rtPCR. These findings suggest that the RPA assay is a simple, rapid, and cost-effective method for PCV-2 detection, which could be potentially applied in clinical diagnosis and field surveillance of PCV-2 infection. © 2016 The Author(s).

Systematic alphanumeric-coded endoscopy versus chromoendoscopy for the detection of precancerous gastric lesions and early gastric cancer in subjects at average risk for gastric cancer.

PubMed

Pérez-Mendoza, A; Zárate-Guzmán, Á M; Galvis García, E S; Sobrino Cossío, S; Djamus Birch, J

Gastric cancer is one of the main causes of cancer worldwide, but there is currently no global screening strategy for the disease. Endoscopy is the screening method of choice in some Asian countries, but no standardized technique has been recognized. Systematic alphanumeric-coded endoscopy can increase gastric lesion detection. The aim of the present article was to compare the usefulness of systematic alphanumeric-coded endoscopy with conventional endoscopy for the detection of premalignant lesions and early gastric cancer in subjects at average risk for gastric cancer. A cross-sectional, comparative, prospective, randomized study was conducted on patients at average risk for gastric cancer (40-50 years of age, no history of H. pylori infection, intestinal metaplasia, gastric atrophy, or gastrointestinal surgery). Before undergoing endoscopy, the patients had gastric preparation (200mg of oral acetylcysteine or 50mg of oral dimethicone). Conventional chromoendoscopy was performed with indigo carmine dye for contrast enhancement. Fifty consecutive cases (mean age 44.4 ± 3.34 years, 60% women, BMI 27.6 ± 5.82 kg/m 2 ) were evaluated. Endoscopic imaging quality was satisfactory in all the cases, with no differences between methods (p = 0.817). The detection rate of premalignant lesions and early gastric cancer was 14% (6 cases of intestinal metaplasia and one case of gastric adenocarcinoma). Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy were 100, 95, 80, 100 and 96%, respectively, for systematic alphanumeric-coded endoscopy, and 100, 45, 20, 100, and 52%, respectively, for conventional endoscopy. Lesion detection through systematic alphanumeric-coded endoscopy was superior to that of conventional endoscopy (p = 0.003; OR = 12). Both techniques were effective, but systematic alphanumeric-coded endoscopy significantly reduced the false positive rate. Copyright © 2018 Asociación Mexicana de Gastroenterología. Publicado por Masson Doyma México S.A. All rights reserved.

Molecular Characterization of cyanobacterial blooms

EPA Science Inventory

Traditionally, the detection and identification of cyanobacteria implicated in harmful algal blooms has been conducted using microscopical techniques. Such conventional methods are time consuming and cumbersome, cannot discriminate between closely related taxa, and cannot discrim...

Intra-laboratory validation of microplate methods for total phenolic content and antioxidant activity on polyphenolic extracts, and comparison with conventional spectrophotometric methods.

PubMed

Bobo-García, Gloria; Davidov-Pardo, Gabriel; Arroqui, Cristina; Vírseda, Paloma; Marín-Arroyo, María R; Navarro, Montserrat

2015-01-01

Total phenolic content (TPC) and antioxidant activity (AA) assays in microplates save resources and time, therefore they can be useful to overcome the fact that the conventional methods are time-consuming, labour intensive and use large amounts of reagents. An intra-laboratory validation of the Folin-Ciocalteu microplate method to measure TPC and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) microplate method to measure AA was performed and compared with conventional spectrophotometric methods. To compare the TPC methods, the confidence intervals of a linear regression were used. In the range of 10-70 mg L(-1) of gallic acid equivalents (GAE), both methods were equivalent. To compare the AA methodologies, the F-test and t-test were used in a range from 220 to 320 µmol L(-1) of Trolox equivalents. Both methods had homogeneous variances, and the means were not significantively different. The limits of detection and quantification for the TPC microplate method were 0.74 and 2.24 mg L(-1) GAE and for the DPPH 12.07 and 36.58 µmol L(-1) of Trolox equivalents. The relative standard deviation of the repeatability and reproducibility for both microplate methods were ≤ 6.1%. The accuracy ranged from 88% to 100%. The microplate and the conventional methods are equals in a 95% confidence level. © 2014 Society of Chemical Industry.

Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma.

PubMed

Flores-Montero, J; Sanoja-Flores, L; Paiva, B; Puig, N; García-Sánchez, O; Böttcher, S; van der Velden, V H J; Pérez-Morán, J-J; Vidriales, M-B; García-Sanz, R; Jimenez, C; González, M; Martínez-López, J; Corral-Mateos, A; Grigore, G-E; Fluxá, R; Pontes, R; Caetano, J; Sedek, L; Del Cañizo, M-C; Bladé, J; Lahuerta, J-J; Aguilar, C; Bárez, A; García-Mateo, A; Labrador, J; Leoz, P; Aguilera-Sanz, C; San-Miguel, J; Mateos, M-V; Durie, B; van Dongen, J J M; Orfao, A

2017-10-01

Flow cytometry has become a highly valuable method to monitor minimal residual disease (MRD) and evaluate the depth of complete response (CR) in bone marrow (BM) of multiple myeloma (MM) after therapy. However, current flow-MRD has lower sensitivity than molecular methods and lacks standardization. Here we report on a novel next generation flow (NGF) approach for highly sensitive and standardized MRD detection in MM. An optimized 2-tube 8-color antibody panel was constructed in five cycles of design-evaluation-redesign. In addition, a bulk-lysis procedure was established for acquisition of ⩾10 7 cells/sample, and novel software tools were constructed for automatic plasma cell gating. Multicenter evaluation of 110 follow-up BM from MM patients in very good partial response (VGPR) or CR showed a higher sensitivity for NGF-MRD vs conventional 8-color flow-MRD -MRD-positive rate of 47 vs 34% (P=0.003)-. Thus, 25% of patients classified as MRD-negative by conventional 8-color flow were MRD-positive by NGF, translating into a significantly longer progression-free survival for MRD-negative vs MRD-positive CR patients by NGF (75% progression-free survival not reached vs 7 months; P=0.02). This study establishes EuroFlow-based NGF as a highly sensitive, fully standardized approach for MRD detection in MM which overcomes the major limitations of conventional flow-MRD methods and is ready for implementation in routine diagnostics.

Nonlethal laparoscopic detection of intersex (testicular oocytes) in largemouthbass (Micropterus salmoides) and smallmouth bass (Micropterus dolomieu)

USGS Publications Warehouse

Blazer, Vicki S.; Macleod, Alexander H; Matsche, Mark A; Yonkos, Lance T

2017-01-01

Intersex in wild fish populations has received considerable attention in the scientific literature and public media. Conventional detection of testicular oocytes (TO), the presence of immature oocytes within testis of male fish, employs transverse sectioning of excised testis and is lethal. This present study used a non-lethal laparoscopic technique to collect biopsies of testis from black bass, entering the body cavity via the genital pore. Detection of TO was compared between biopsy and conventional methods using 79 smallmouth bass (SMB) Micropterus dolomieu from 8 sites and 68 largemouth bass (LMB) M. salmoides from 4 sites. Both methods performed similarly at sites where TO severity was moderate or high (6 of 8 SMB sites) while transverse sectioning resulted in superior TO detection at sites where severity was low (2 of 8 SMB sites and all 4 LMB sites). In SMB, TO prevalence by transverse and biopsy methods was strongly correlated across sites (r2 = 0.81) and severity reported by enumeration of TO was moderately correlated across sites (r2 = 0.59). Survival of a subset of LMB (n = 20) to 28-d after laparoscopic surgery was 90%. This research indicates that laparoscopy may be useful for monitoring the prevalence and severity of TO in Micropterus species, particularly when lethal sampling is precluded.

Rapid and Highly Sensitive Detection of Lead Ions in Drinking Water Based on a Strip Immunosensor

PubMed Central

Kuang, Hua; Xing, Changrui; Hao, Changlong; Liu, Liqiang; Wang, Libing; Xu, Chuanlai

2013-01-01

In this study, we have first developed a rapid and sensitive strip immunosensor based on two heterogeneously-sized gold nanoparticles (Au NPs) probes for the detection of trace lead ions in drinking water. The sensitivity was 4-fold higher than that of the conventional LFA under the optimized conditions. The visual limit of detection (LOD) of the amplified method for qualitative detection lead ions was 2 ng/mL and the LOD for semi-quantitative detection could go down to 0.19 ng/mL using a scanning reader. The method suffered from no interference from other metal ions and could be used to detect trace lead ions in drinking water without sample enrichment. The recovery of the test samples ranged from 96% to 103%. As the detection method could be accomplished within 15 min, this method could be used as a potential tool for preliminary monitoring of lead contamination in drinking water. PMID:23539028

Detection of cucumber green mottle mosaic virus-infected watermelon seeds using short wave infrared (SWIR) hyperspectral imaging system

USDA-ARS?s Scientific Manuscript database

The cucurbit diseases caused by cucumber green mottle mosaic virus (CGMMV) have led to a serious problem to growers and seed producers because it is difficult to prevent spreading through causal agent of seeds. Conventional detection methods for infected seed such as a biological, serological, and m...

Searching for life in the Universe: unconventional methods for an unconventional problem.

PubMed

Nealson, K H; Tsapin, A; Storrie-Lombardi, M

2002-12-01

The search for life, on and off our planet, can be done by conventional methods with which we are all familiar. These methods are sensitive and specific, and are often capable of detecting even single cells. However, if the search broadens to include life that may be different (even subtly different) in composition, the methods and even the approach must be altered. Here we discuss the development of what we call non-earthcentric life detection--detecting life with methods that could detect life no matter what its form or composition. To develop these methods, we simply ask, can we define life in terms of its general properties and particularly those that can be measured and quantified? Taking such an approach we can search for life using physics and chemistry to ask questions about structure, chemical composition, thermodynamics, and kinetics. Structural complexity can be searched for using computer algorithms that recognize complex structures. Once identified, these structures can be examined for a variety of chemical traits, including elemental composition, chirality, and complex chemistry. A second approach involves defining our environment in terms of energy sources (i.e., reductants), and oxidants (e.g. what is available to eat and breathe), and then looking for areas in which such phenomena are inexplicably out of chemical equilibrium. These disequilibria, when found, can then be examined in detail for the presence of the structural and chemical complexity that presumably characterizes any living systems. By this approach, we move the search for life to one that should facilitate the detection of any earthly life it encountered, as well as any non-conventional life forms that have structure, complex chemistry, and live via some form of redox chemistry.

Assessment of a Pan-Dermatophyte Nested-PCR Compared with Conventional Methods for Direct Detection and Identification of Dermatophytosis Agents in Animals.

PubMed

Piri, Fahimeh; Zarei Mahmoudabadi, Ali; Ronagh, Ali; Ahmadi, Bahram; Makimura, Koichi; Rezaei-Matehkolaei, Ali

2018-06-26

Conventional direct microscopy with potassium hydroxide (KOH) and culture were found to lack the ability to establish a fast and specific diagnosis of dermatophytosis. A pan-dermatophyte nested-PCR assay was developed using a novel primer pair targeting the translation elongation factor 1-α (Tef-1α) sequences for direct detection and identification of most veterinary relevant dermatophytes in animal samples suspected to dermatophytosis. A total of 140 animal skin and hair samples were subjected to direct microscopy, culture, and ITS-RFLP/ITS-sequencing of culture isolates for the detection and identification of dermatophytosis agents. Nested-PCR sequencing was performed on all the extracted DNAs using a commercial kit after dissolving the specimens by mechanical beating. Nested-PCR was positive in 90% of samples, followed by direct microscopy (85.7%) and culture (75%). The degree of agreement between nested-PCR and direct microscopy (94.4%) was higher than with culture (83.3%). In 105 culture positive cases, the measures of agreement for the identification of dermatophytosis agents were as follows: 100% between nested-PCR sequencing and ITS-RFLP/ITS-sequencing and 63.8% between nested-PCR sequencing and culture. The developed nested-PCR was faster as well as more sensitive and specific than conventional methods for detection and identification of dermatophytes in clinical samples, which was particularly suitable for epidemiological studies. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Development of a nondestructive leak testing method utilizing the head space analyzer for ampoule products containing ethanol-based solutions.

PubMed

Sudo, Hirotaka; O'driscoll, Michael; Nishiwaki, Kenji; Kawamoto, Yuji; Gammell, Philip; Schramm, Gerhard; Wertli, Toni; Prinz, Heino; Mori, Atsuhide; Sako, Kazuhiro

2012-01-01

The application of a head space analyzer for oxygen concentration was examined to develop a novel ampoule leak test method. Studies using ampoules filled with ethanol-based solution and with nitrogen in the headspace demonstrated that the head space analysis (HSA) method showed sufficient sensitivity in detecting an ampoule crack. The proposed method is the use of HSA in conjunction with the pretreatment of an overpressurising process known as bombing to facilitate the oxygen flow through the crack in the ampoule. The method was examined in comparative studies with a conventional dye ingress method, and the results showed that the HSA method exhibits sensitivity superior to the dye method. The results indicate that the HSA method in combination with the bombing treatment provides potential application as a leak test for the detection of container defects not only for ampoule products with ethanol-based solutions, but also for testing lyophilized products in vials with nitrogen in the head space. The application of a head space analyzer for oxygen concentration was examined to develop a novel ampoule leak test method. The proposed method is the use of head space analysis (HSA) in conjunction with the pretreatment of an overpressurising process known as bombing to facilitate oxygen flow through the crack in the ampoule for use in routine production. The result of the comparative study with a conventional dye leak test method indicates that the HSA method in combination with the bombing treatment can be used as a leak test method, enabling detection of container defects.

Investigation of recrystallization of amorphous trehalose through hot-humidity stage X-ray powder diffraction.

PubMed

Jójárt-Laczkovich, Orsolya; Katona, Gábor; Aigner, Zoltán; Szabó-Révész, Piroska

2016-12-01

The aim of this work was an investigation of the physical changes of the amorphous model material spray-dried trehalose through the use of various analytical techniques and to identify a suitable, rapid method able to quantify the changes. The crystallinity changes and recrystallization process of amorphous samples were investigated by hot-humidity stage X-ray powder diffractometry (HH-XRPD) with fresh samples, conventional X-ray powder diffractometry (XRPD) used stored samples and by differential scanning calorimetry (DSC). The data from the three methods were compared and the various forms of trehalose were analysed. HH-XRPD demonstrated that the recrystallization began at 40 and 60°C up to 45% RH and at 70°C up to 30% RH into dihydrate form. At 70°C up to 60% RH the anhydrous form of trehalose appeared too. Conventional XRPD results showed, that in the 28days stored samples the dihydrate form was detected at 40°C, 50% RH. Storage at 60°C, 40% RH resulted in the appearance of the anhydrous form and at 60°C, 50% RH both polymorphic forms were detected. By carrying out the DSC measurements at different temperatures the fraction of recrystallized trehalose dihydrate was detected. The recrystallization investigated by HH-XRPD and DSC followed Avrami kinetics, the calculated rate constants of isothermal crystallization (K) were same. Both HH-XRPD and conventional XRPD was suitable for the detection of the physical changes of the amorphous model material. DSC measurements showed similar results as HH-XRPD. Primarily HH-XRPD could be suggested for prediction, because the method is fast and every changes could be studied on one sample. Copyright © 2016 Elsevier B.V. All rights reserved.

Extrapolation-Based References Improve Motion and Eddy-Current Correction of High B-Value DWI Data: Application in Parkinson's Disease Dementia.

PubMed

Nilsson, Markus; Szczepankiewicz, Filip; van Westen, Danielle; Hansson, Oskar

2015-01-01

Conventional motion and eddy-current correction, where each diffusion-weighted volume is registered to a non diffusion-weighted reference, suffers from poor accuracy for high b-value data. An alternative approach is to extrapolate reference volumes from low b-value data. We aim to compare the performance of conventional and extrapolation-based correction of diffusional kurtosis imaging (DKI) data, and to demonstrate the impact of the correction approach on group comparison studies. DKI was performed in patients with Parkinson's disease dementia (PDD), and healthy age-matched controls, using b-values of up to 2750 s/mm2. The accuracy of conventional and extrapolation-based correction methods was investigated. Parameters from DTI and DKI were compared between patients and controls in the cingulum and the anterior thalamic projection tract. Conventional correction resulted in systematic registration errors for high b-value data. The extrapolation-based methods did not exhibit such errors, yielding more accurate tractography and up to 50% lower standard deviation in DKI metrics. Statistically significant differences were found between patients and controls when using the extrapolation-based motion correction that were not detected when using the conventional method. We recommend that conventional motion and eddy-current correction should be abandoned for high b-value data in favour of more accurate methods using extrapolation-based references.

Beyond Blood Culture and Gram Stain Analysis: A Review of Molecular Techniques for the Early Detection of Bacteremia in Surgical Patients.

PubMed

Scerbo, Michelle H; Kaplan, Heidi B; Dua, Anahita; Litwin, Douglas B; Ambrose, Catherine G; Moore, Laura J; Murray, Col Clinton K; Wade, Charles E; Holcomb, John B

2016-06-01

Sepsis from bacteremia occurs in 250,000 cases annually in the United States, has a mortality rate as high as 60%, and is associated with a poorer prognosis than localized infection. Because of these high figures, empiric antibiotic administration for patients with systemic inflammatory response syndrome (SIRS) and suspected infection is the second most common indication for antibiotic administration in intensive care units (ICU)s. However, overuse of empiric antibiotics contributes to the development of opportunistic infections, antibiotic resistance, and the increase in multi-drug-resistant bacterial strains. The current method of diagnosing and ruling out bacteremia is via blood culture (BC) and Gram stain (GS) analysis. Conventional and molecular methods for diagnosing bacteremia were reviewed and compared. The clinical implications, use, and current clinical trials of polymerase chain reaction (PCR)-based methods to detect bacterial pathogens in the blood stream were detailed. BC/GS has several disadvantages. These include: some bacteria do not grow in culture media; others do not GS appropriately; and cultures can require up to 5 d to guide or discontinue antibiotic treatment. PCR-based methods can be potentially applied to detect rapidly, accurately, and directly microbes in human blood samples. Compared with the conventional BC/GS, particular advantages to molecular methods (specifically, PCR-based methods) include faster results, leading to possible improved antibiotic stewardship when bacteremia is not present.

Fractal analysis of GPS time series for early detection of disastrous seismic events

NASA Astrophysics Data System (ADS)

Filatov, Denis M.; Lyubushin, Alexey A.

2017-03-01

A new method of fractal analysis of time series for estimating the chaoticity of behaviour of open stochastic dynamical systems is developed. The method is a modification of the conventional detrended fluctuation analysis (DFA) technique. We start from analysing both methods from the physical point of view and demonstrate the difference between them which results in a higher accuracy of the new method compared to the conventional DFA. Then, applying the developed method to estimate the measure of chaoticity of a real dynamical system - the Earth's crust, we reveal that the latter exhibits two distinct mechanisms of transition to a critical state: while the first mechanism has already been known due to numerous studies of other dynamical systems, the second one is new and has not previously been described. Using GPS time series, we demonstrate efficiency of the developed method in identification of critical states of the Earth's crust. Finally we employ the method to solve a practically important task: we show how the developed measure of chaoticity can be used for early detection of disastrous seismic events and provide a detailed discussion of the numerical results, which are shown to be consistent with outcomes of other researches on the topic.

Diagnostic accuracy of susceptibility-weighted magnetic resonance imaging for the evaluation of pineal gland calcification

PubMed Central

Böker, Sarah M.; Bender, Yvonne Y.; Diederichs, Gerd; Fallenberg, Eva M.; Wagner, Moritz; Hamm, Bernd; Makowski, Marcus R.

2017-01-01

Objectives To determine the diagnostic performance of susceptibility-weighted magnetic resonance imaging (SWMR) for the detection of pineal gland calcifications (PGC) compared to conventional magnetic resonance imaging (MRI) sequences, using computed tomography (CT) as a reference standard. Methods 384 patients who received a 1.5 Tesla MRI scan including SWMR sequences and a CT scan of the brain between January 2014 and October 2016 were retrospectively evaluated. 346 patients were included in the analysis, of which 214 showed PGC on CT scans. To assess correlation between imaging modalities, the maximum calcification diameter was used. Sensitivity and specificity and intra- and interobserver reliability were calculated for SWMR and conventional MRI sequences. Results SWMR reached a sensitivity of 95% (95% CI: 91%-97%) and a specificity of 96% (95% CI: 91%-99%) for the detection of PGC, whereas conventional MRI achieved a sensitivity of 43% (95% CI: 36%-50%) and a specificity of 96% (95% CI: 91%-99%). Detection rates for calcifications in SWMR and conventional MRI differed significantly (95% versus 43%, p<0.001). Diameter measurements between SWMR and CT showed a close correlation (R2 = 0.85, p<0.001) with a slight but not significant overestimation of size (SWMR: 6.5 mm ± 2.5; CT: 5.9 mm ± 2.4, p = 0.02). Interobserver-agreement for diameter measurements was excellent on SWMR (ICC = 0.984, p < 0.0001). Conclusions Combining SWMR magnitude and phase information enables the accurate detection of PGC and offers a better diagnostic performance than conventional MRI with CT as a reference standard. PMID:28278291

[Role of BoBs technology in early missed abortion chorionic villi].

PubMed

Li, Z Y; Liu, X Y; Peng, P; Chen, N; Ou, J; Hao, N; Zhou, J; Bian, X M

2018-05-25

Objective: To investigate the value of bacterial artificial chromosome-on-beads (BoBs) technology in the genetic analysis of early missed abortion chorionic villi. Methods: Early missed abortion chorionic villi were detected with both conventional karyotyping method and BoBs technology in Peking Union Medical Hospital from July 2014 to March 2015. Compared the results of BoBs with conventional karyotyping analysis to evaluate the sensitivity, specificity and accuracy of this new method. Results: (1) A total of 161 samples were tested successfully in the technology of BoBs, 131 samples were tested successfully in the method of conventional karyotyping. (2) All of the cases obtained from BoBs results in (2.7±0.6) days and obtained from conventional karyotyping results in (22.5±1.9) days. There was significant statistical difference between the two groups ( t= 123.315, P< 0.01) . (3) Out of 161 cases tested in BoBs, 85 (52.8%, 85/161) cases had the abnormal chromosomes, including 79 cases chromosome number abnormality, 4 cases were chromosome segment deletion, 2 cases mosaic. Out of 131 cases tested successfully in conventional karyotyping, 79 (60.3%, 79/131) cases had the abnormal chromosomes including 62 cases chromosome number abnormality, 17 cases other chromosome number abnormality, and the rate of chromosome abnormality between two methods was no significant differences ( P =0.198) . (4) Conventional karyotyping results were served as the gold standard, the accuracy of BoBs for abnormal chromosomes was 82.4% (108/131) , analysed the normal chromosomes (52 cases) and chromosome number abnormality (62 cases) tested in conventional karyotyping, the accuracy of BoBs for chromosome number abnormality was 94.7% (108/114) . Conclusion: BoBs is a rapid reliable and easily operated method to test early missed abortion chorionic villi chromosomal abnormalities.

Evaluation of microplate immunocapture method for detection of Vibrio cholerae, Salmonella Typhi and Shigella flexneri from food.

PubMed

Fakruddin, Md; Hossain, Md Nur; Ahmed, Monzur Morshed

2017-08-29

Improved methods with better separation and concentration ability for detection of foodborne pathogens are in constant need. The aim of this study was to evaluate microplate immunocapture (IC) method for detection of Salmonella Typhi, Shigella flexneri and Vibrio cholerae from food samples to provide a better alternative to conventional culture based methods. The IC method was optimized for incubation time, bacterial concentration, and capture efficiency. 6 h incubation and log 6 CFU/ml cell concentration provided optimal results. The method was shown to be highly specific for the pathogens concerned. Capture efficiency (CE) was around 100% of the target pathogens, whereas CE was either zero or very low for non-target pathogens. The IC method also showed better pathogen detection ability at different concentrations of cells from artificially contaminated food samples in comparison with culture based methods. Performance parameter of the method was also comparable (Detection limit- 25 CFU/25 g; sensitivity 100%; specificity-96.8%; Accuracy-96.7%), even better than culture based methods (Detection limit- 125 CFU/25 g; sensitivity 95.9%; specificity-97%; Accuracy-96.2%). The IC method poses to be the potential to be used as a method of choice for detection of foodborne pathogens in routine laboratory practice after proper validation.

Methods for Joining of Rails : Survey Report

DOT National Transportation Integrated Search

1977-07-01

The performance of track structures depends greatly on the integrity of the connections between rail sections. Because the majority of service and detected rail failures occur at joints, particularly conventional bolted joints, this survey was conduc...

Determination of Trace Available Heavy Metals in Soil Using Laser-Induced Breakdown Spectroscopy Assisted with Phase Transformation Method.

PubMed

Yi, Rongxing; Yang, Xinyan; Zhou, Ran; Li, Jiaming; Yu, Huiwu; Hao, Zhongqi; Guo, Lianbo; Li, Xiangyou; Lu, Yongfeng; Zeng, Xiaoyan

2018-05-18

To detect available heavy metals in soil using laser-induced breakdown spectroscopy (LIBS) and improve its poor detection sensitivity, a simple and low cost sample pretreatment method named solid-liquid-solid transformation was proposed. By this method, available heavy metals were extracted from soil through ultrasonic vibration and centrifuging and then deposited on a glass slide. Utilization of this solid-liquid-solid transformation method, available Cd and Pb elements in soil were detected successfully. The results show that the regression coefficients of calibration curves for soil analyses reach to more than 0.98. The limits of detection could reach to 0.067 and 0.94 ppm for available Cd and Pb elements in soil under optimized conditions, respectively, which are much better than those obtained by conventional LIBS.

Real-time traffic sign recognition based on a general purpose GPU and deep-learning.

PubMed

Lim, Kwangyong; Hong, Yongwon; Choi, Yeongwoo; Byun, Hyeran

2017-01-01

We present a General Purpose Graphics Processing Unit (GPGPU) based real-time traffic sign detection and recognition method that is robust against illumination changes. There have been many approaches to traffic sign recognition in various research fields; however, previous approaches faced several limitations when under low illumination or wide variance of light conditions. To overcome these drawbacks and improve processing speeds, we propose a method that 1) is robust against illumination changes, 2) uses GPGPU-based real-time traffic sign detection, and 3) performs region detecting and recognition using a hierarchical model. This method produces stable results in low illumination environments. Both detection and hierarchical recognition are performed in real-time, and the proposed method achieves 0.97 F1-score on our collective dataset, which uses the Vienna convention traffic rules (Germany and South Korea).

Improved automated lumen contour detection by novel multifrequency processing algorithm with current intravascular ultrasound system.

PubMed

Kume, Teruyoshi; Kim, Byeong-Keuk; Waseda, Katsuhisa; Sathyanarayana, Shashidhar; Li, Wenguang; Teo, Tat-Jin; Yock, Paul G; Fitzgerald, Peter J; Honda, Yasuhiro

2013-02-01

The aim of this study was to evaluate a new fully automated lumen border tracing system based on a novel multifrequency processing algorithm. We developed the multifrequency processing method to enhance arterial lumen detection by exploiting the differential scattering characteristics of blood and arterial tissue. The implementation of the method can be integrated into current intravascular ultrasound (IVUS) hardware. This study was performed in vivo with conventional 40-MHz IVUS catheters (Atlantis SR Pro™, Boston Scientific Corp, Natick, MA) in 43 clinical patients with coronary artery disease. A total of 522 frames were randomly selected, and lumen areas were measured after automatically tracing lumen borders with the new tracing system and a commercially available tracing system (TraceAssist™) referred to as the "conventional tracing system." The data assessed by the two automated systems were compared with the results of manual tracings by experienced IVUS analysts. New automated lumen measurements showed better agreement with manual lumen area tracings compared with those of the conventional tracing system (correlation coefficient: 0.819 vs. 0.509). When compared against manual tracings, the new algorithm also demonstrated improved systematic error (mean difference: 0.13 vs. -1.02 mm(2) ) and random variability (standard deviation of difference: 2.21 vs. 4.02 mm(2) ) compared with the conventional tracing system. This preliminary study showed that the novel fully automated tracing system based on the multifrequency processing algorithm can provide more accurate lumen border detection than current automated tracing systems and thus, offer a more reliable quantitative evaluation of lumen geometry. Copyright © 2011 Wiley Periodicals, Inc.

Performance of a high-sensitivity dedicated cardiac SPECT scanner for striatal uptake quantification in the brain based on analysis of projection data

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

Park, Mi-Ae; Moore, Stephen C.; McQuaid, Sarah J.

Purpose: The authors have previously reported the advantages of high-sensitivity single-photon emission computed tomography (SPECT) systems for imaging structures located deep inside the brain. DaTscan (Isoflupane I-123) is a dopamine transporter (DaT) imaging agent that has shown potential for early detection of Parkinson disease (PD), as well as for monitoring progression of the disease. Realizing the full potential of DaTscan requires efficient estimation of striatal uptake from SPECT images. They have evaluated two SPECT systems, a conventional dual-head gamma camera with low-energy high-resolution collimators (conventional) and a dedicated high-sensitivity multidetector cardiac imaging system (dedicated) for imaging tasks related to PD.more » Methods: Cramer-Rao bounds (CRB) on precision of estimates of striatal and background activity concentrations were calculated from high-count, separate acquisitions of the compartments (right striata, left striata, background) of a striatal phantom. CRB on striatal and background activity concentration were calculated from essentially noise-free projection datasets, synthesized by scaling and summing the compartment projection datasets, for a range of total detected counts. They also calculated variances of estimates of specific-to-nonspecific binding ratios (BR) and asymmetry indices from these values using propagation of error analysis, as well as the precision of measuring changes in BR on the order of the average annual decline in early PD. Results: Under typical clinical conditions, the conventional camera detected 2 M counts while the dedicated camera detected 12 M counts. Assuming a normal BR of 5, the standard deviation of BR estimates was 0.042 and 0.021 for the conventional and dedicated system, respectively. For an 8% decrease to BR = 4.6, the signal-to-noise ratio were 6.8 (conventional) and 13.3 (dedicated); for a 5% decrease, they were 4.2 (conventional) and 8.3 (dedicated). Conclusions: This implies that PD can be detected earlier with the dedicated system than with the conventional system; therefore, earlier identification of PD progression should be possible with the high-sensitivity dedicated SPECT camera.« less

Implementation of a novel efficient low cost method in structural health monitoring

NASA Astrophysics Data System (ADS)

Asadi, S.; Sepehry, N.; Shamshirsaz, M.; Vaghasloo, Y. A.

2017-05-01

In active structural health monitoring (SHM) methods, it is necessary to excite the structure with a preselected signal. More studies in the field of active SHM are focused on applying SHM on higher frequency ranges since it is possible to detect smaller damages, using higher excitation frequency. Also, to increase spatial domain of measurements and enhance signal to noise ratio (SNR), the amplitude of excitation signal is usually amplified. These issues become substantial where piezoelectric transducers with relatively high capacitance are used and consequently, need to utilize high power amplifiers becomes predominant. In this paper, a novel method named Step Excitation Method (SEM) is proposed and implemented for Lamb wave and transfer impedance-based SHM for damage detection in structures. Three different types of structure are studied: beam, plate and pipe. The related hardware is designed and fabricated which eliminates high power analog amplifiers and decreases complexity of driver significantly. Spectral Finite Element Method (SFEM) is applied to examine performance of proposed SEM. In proposed method, by determination of impulse response of the system, any input could be applied to the system by both finite element simulations and experiments without need for multiple measurements. The experimental results using SEM are compared with those obtained by conventional direct excitation method for healthy and damaged structures. The results show an improvement of amplitude resolution in damage detection comparing to conventional method which is due to achieving an SNR improvement up to 50%.

Tests of various colorants for application of a Fourier transform infrared imaging system to deciphering obliterated writings

NASA Astrophysics Data System (ADS)

Sugawara, Shigeru

2015-10-01

Obliterated writing is writing that has been obscured by different-colored materials. There are obliterated writings that cannot be detected by conventional methods. A method for deciphering such obliterated writings was developed in this study. Mid-infrared spectroscopic imaging in the wavelength range of 2.5-14 μm was used for deciphering because the infrared spectrum differs among different brands of colorants. Obliterated writings were made by pressing information protection stamps onto characters written by 4 kinds of colorants. The samples were tested for deciphering by the Fourier-transform infrared imaging system. Two peak areas of two specific wavenumber regions of each reflectance spectrum were calculated and the ratio of the two values is displayed as a unique gray scale in the spectroscopic image. As a result, the absorption peak at various wavenumbers could be used to decipher obliterated writings that could not be detected by the conventional methods. Ten different parameters for deciphering obliterated writing were found in this study.

Tick-Borne Pathogen – Reversed and Conventional Discovery of Disease

PubMed Central

Tijsse-Klasen, Ellen; Koopmans, Marion P. G.; Sprong, Hein

2014-01-01

Molecular methods have increased the number of known microorganisms associated with ticks significantly. Some of these newly identified microorganisms are readily linked to human disease while others are yet unknown to cause human disease. The face of tick-borne disease discovery has changed with more diseases now being discovered in a “reversed way,” detecting disease cases only years after the tick-borne microorganism was first discovered. Compared to the conventional discovery of infectious diseases, reverse order discovery presents researchers with new challenges. Estimating public health risks of such agents is especially challenging, as case definitions and diagnostic procedures may initially be missing. We discuss the advantages and shortcomings of molecular methods, serology, and epidemiological studies that might be used to study some fundamental questions regarding newly identified tick-borne diseases. With increased tick-exposure and improved detection methods, more tick-borne microorganisms will be added to the list of pathogens causing disease in humans in the future. PMID:25072045

Laser-based pedestrian tracking in outdoor environments by multiple mobile robots.

PubMed

Ozaki, Masataka; Kakimuma, Kei; Hashimoto, Masafumi; Takahashi, Kazuhiko

2012-10-29

This paper presents an outdoors laser-based pedestrian tracking system using a group of mobile robots located near each other. Each robot detects pedestrians from its own laser scan image using an occupancy-grid-based method, and the robot tracks the detected pedestrians via Kalman filtering and global-nearest-neighbor (GNN)-based data association. The tracking data is broadcast to multiple robots through intercommunication and is combined using the covariance intersection (CI) method. For pedestrian tracking, each robot identifies its own posture using real-time-kinematic GPS (RTK-GPS) and laser scan matching. Using our cooperative tracking method, all the robots share the tracking data with each other; hence, individual robots can always recognize pedestrians that are invisible to any other robot. The simulation and experimental results show that cooperating tracking provides the tracking performance better than conventional individual tracking does. Our tracking system functions in a decentralized manner without any central server, and therefore, this provides a degree of scalability and robustness that cannot be achieved by conventional centralized architectures.

Application of loop-mediated isothermal amplification (LAMP) assay for the rapid diagnosis of pathogenic bacteria in clinical sputum specimens of acute exacerbation of COPD (AECOPD).

PubMed

Zhang, Wei; Chen, Chuanhui; Cui, Jian; Bai, Wei; Zhou, Jing

2015-01-01

The present study explores the application of LAMP for rapid diagnosis of pathogenic bacteria in clinical sputum specimens of AECOPD as compared with conventional sputum culturing method. 120 sputum specimens of AECOPD patients, 46 sputum specimens of healthy controls, as well as 166 serum specimens as negative controls, were evaluated by LAMP assay using primers of eight typical respiratory pathogens. No cross-reactivity was observed in these negative control species using LAMP assay. The lower detection limit of LAMP assay was approximately 10(3) copies. 25 cases (20.8%) were detected at least one positive bacteria species by conventional sputum culturing method, while 73 cases (60.8%) were tested positive in LAMP assay. Moreover, compared with sputum culture, bacterial titers results of LAMP assay were more consistent with FEV1/FVC value of AECOPD patients. These results indicated that the sensitivity of LAMP assay was significantly higher than that of sputum culturing method.

Convenient Detection of the Citrus Greening (Huanglongbing) Bacterium ‘Candidatus Liberibacter asiaticus’ by Direct PCR from the Midrib Extract

PubMed Central

Fujikawa, Takashi; Miyata, Shin-Ichi; Iwanami, Toru

2013-01-01

A phloem-limited bacterium, ‘Candidatus Liberibacter asiaticus’ (Las) is a major pathogen of citrus greening (huanglongbing), one of the most destructive citrus diseases worldwide. The rapid identification and culling of infected trees and budwoods in quarantine are the most important control measures. DNA amplification including conventional polymerase chain reaction (PCR) has commonly been used for rapid detection and identification. However, long and laborious procedures for DNA extraction have greatly reduced the applicability of this method. In this study, we found that the Las bacterial cells in the midribs of infected leaves were extracted rapidly and easily by pulverization and centrifugation with mini homogenization tubes. We also found that the Las bacterial cells in the midrib extract were suitable for highly sensitive direct PCR. The performance of direct PCR using this extraction method was not inferior to that of conventional PCR. Thus, the direct PCR method described herein is characterized by its simplicity, sensitivity, and robustness, and is applicable to quarantine testing. PMID:23437295

Quasi-closed phase forward-backward linear prediction analysis of speech for accurate formant detection and estimation.

PubMed

Gowda, Dhananjaya; Airaksinen, Manu; Alku, Paavo

2017-09-01

Recently, a quasi-closed phase (QCP) analysis of speech signals for accurate glottal inverse filtering was proposed. However, the QCP analysis which belongs to the family of temporally weighted linear prediction (WLP) methods uses the conventional forward type of sample prediction. This may not be the best choice especially in computing WLP models with a hard-limiting weighting function. A sample selective minimization of the prediction error in WLP reduces the effective number of samples available within a given window frame. To counter this problem, a modified quasi-closed phase forward-backward (QCP-FB) analysis is proposed, wherein each sample is predicted based on its past as well as future samples thereby utilizing the available number of samples more effectively. Formant detection and estimation experiments on synthetic vowels generated using a physical modeling approach as well as natural speech utterances show that the proposed QCP-FB method yields statistically significant improvements over the conventional linear prediction and QCP methods.

Determination of mutagenic amines in water and food samples by high pressure liquid chromatography with amperometric detection using a multiwall carbon nanotubes-glassy carbon electrode.

PubMed

Bueno, Ana María; Marín, Miguel Ángel; Contento, Ana María; Ríos, Ángel

2016-02-01

A chromatographic method, using amperometric detection, for the sensitive determination of six representative mutagenic amines was developed. A glassy carbon electrode (GCE), modified with multiwall carbon nanotubes (GCE-CNTs), was prepared and its response compared to a conventional glassy carbon electrode. The chromatographic method (HPLC-GCE-CNTs) allowed the separation and the determination of heterocyclic aromatic amines (HAAs) classified as mutagenic amines by the International Agency for Research of Cancer. The new electrode was systematically studied in terms of stability, sensitivity, and reproducibility. Statistical analysis of the obtained data demonstrated that the modified electrode provided better sensitivity than the conventional unmodified ones. Detection limits were in the 3.0 and 7.5 ng/mL range, whereas quantification limits ranged between 9.5 and 25.0 ng/mL were obtained. The applicability of the method was demonstrated by the determination of the amines in several types of samples (water and food samples). Recoveries indicate very good agreement between amounts added and those found for all HAAs (recoveries in the 92% and 105% range). Copyright © 2015 Elsevier Ltd. All rights reserved.

Synthetic Hounsfield units from spectral CT data

NASA Astrophysics Data System (ADS)

Bornefalk, Hans

2012-04-01

Beam-hardening-free synthetic images with absolute CT numbers that radiologists are used to can be constructed from spectral CT data by forming ‘dichromatic’ images after basis decomposition. The CT numbers are accurate for all tissues and the method does not require additional reconstruction. This method prevents radiologists from having to relearn new rules-of-thumb regarding absolute CT numbers for various organs and conditions as conventional CT is replaced by spectral CT. Displaying the synthetic Hounsfield unit images side-by-side with images reconstructed for optimal detectability for a certain task can ease the transition from conventional to spectral CT.

Rapid detection of mecA and spa by the loop-mediated isothermal amplification (LAMP) method.

PubMed

Koide, Y; Maeda, H; Yamabe, K; Naruishi, K; Yamamoto, T; Kokeguchi, S; Takashiba, S

2010-04-01

To develop a detection assay for staphylococcal mecA and spa by using loop-mediated isothermal amplification (LAMP) method. Staphylococcus aureus and other related species were subjected to the detection of mecA and spa by both PCR and LAMP methods. The LAMP successfully amplified the genes under isothermal conditions at 64 degrees C within 60 min, and demonstrated identical results with the conventional PCR methods. The detection limits of the LAMP for mecA and spa, by gel electrophoresis, were 10(2) and 10 cells per tube, respectively. The naked-eye inspections were possible with 10(3) and 10 cells for detection of mecA and spa, respectively. The LAMP method was then applied to sputum and dental plaque samples. The LAMP and PCR demonstrated identical results for the plaque samples, although frequency in detection of mecA and spa by the LAMP was relatively lower for the sputum samples when compared to the PCR methods. Application of the LAMP enabled a rapid detection assay for mecA and spa. The assay may be applicable to clinical plaque samples. The LAMP offers an alternative detection assay for mecA and spa with a great advantage of the rapidity.

Thermoelectric SQUID method for the detection of segregations

NASA Astrophysics Data System (ADS)

Hinken, Johann H.; Tavrin, Yury

2000-05-01

Aero engine turbine discs are most critical parts. Material inhomogeneities can cause disc fractures during the flight with fatal air disasters. Nondestructive testing (NDT) of the discs in various machining steps is necessary and performed as well as possible. Conventional NDT methods, however, like eddy current testing and ultrasonic testing have unacceptable limits. For example, subsurface segregations often cannot be detected directly but only indirectly in such cases when cracks already have developed from them. This may be too late. A new NDT method, which we call the Thermoelectric SQUID Method, has been developed. It allows for the detection of metallic inclusions within non-ferromagnetic metallic base material. This paper describes the results of a feasibility study on aero engine turbine discs made from Inconel® 718. These contained segregations that had been detected before by anodic etching. With the Thermoelectric SQUID Method, these segregations were detected again, and further segregations below the surfaces have been found, which had not been detected before. For this new NDT method the disc material is quasi-transparent. The Thermoelectric SQUID Method is also useful to detect distributed and localized inhomogeneities in pure metals like niobium sheets for particle accelerators.

A Support System for Mouse Operations Using Eye-Gaze Input

NASA Astrophysics Data System (ADS)

Abe, Kiyohiko; Nakayama, Yasuhiro; Ohi, Shoichi; Ohyama, Minoru

We have developed an eye-gaze input system for people with severe physical disabilities, such as amyotrophic lateral sclerosis (ALS) patients. This system utilizes a personal computer and a home video camera to detect eye-gaze under natural light. The system detects both vertical and horizontal eye-gaze by simple image analysis, and does not require special image processing units or sensors. Our conventional eye-gaze input system can detect horizontal eye-gaze with a high degree of accuracy. However, it can only classify vertical eye-gaze into 3 directions (up, middle and down). In this paper, we propose a new method for vertical eye-gaze detection. This method utilizes the limbus tracking method for vertical eye-gaze detection. Therefore our new eye-gaze input system can detect the two-dimension coordinates of user's gazing point. By using this method, we develop a new support system for mouse operation. This system can move the mouse cursor to user's gazing point.

Continuous Blood Pressure Monitoring in Daily Life

NASA Astrophysics Data System (ADS)

Lopez, Guillaume; Shuzo, Masaki; Ushida, Hiroyuki; Hidaka, Keita; Yanagimoto, Shintaro; Imai, Yasushi; Kosaka, Akio; Delaunay, Jean-Jacques; Yamada, Ichiro

Continuous monitoring of blood pressure in daily life could improve early detection of cardiovascular disorders, as well as promoting healthcare. Conventional ambulatory blood pressure monitoring (ABPM) equipment can measure blood pressure at regular intervals for 24 hours, but is limited by long measuring time, low sampling rate, and constrained measuring posture. In this paper, we demonstrate a new method for continuous real-time measurement of blood pressure during daily activities. Our method is based on blood pressure estimation from pulse wave velocity (PWV) calculation, which formula we improved to take into account changes in the inner diameter of blood vessels. Blood pressure estimation results using our new method showed a greater precision of measured data during exercise, and a better accuracy than the conventional PWV method.

[Evaluation of four methods for detecting methicillin-resistant Staphylococcus aureus isolates from clinical specimens at a regional hospital in Mexico].

PubMed

Acosta-Pérez, Gabriel; Rodríguez-Ábrego, Gabriela; Longoria-Revilla, Ernesto; Castro-Mussot, María Eugenia

2012-01-01

To estimate the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in clinical isolates and to compare different methods for detection of MRSA in a lab with limited available personnel and resources. 140 Staphylococcus aureus strains isolated from patients in several departments were assayed for β-lactamase production, MIC-Vitek 2 oxacillin, ChromID MRSA, disk diffusion in agar for cefoxitin 30 μg and PBP2a detection. The results of conventional tests were compared with the "gold standard" PCR test for mecA gene. Cohen´s kappa index was also calculated in order to evaluate the intra assay agreement between the used methods. The found prevalence was 90.7%. Sensitivity and specificity were: disk diffusion for cefoxitin 97 and 92% respectively, MIC Vitek 2-XL 97 and 69%, ChromoID MRSA 97 and 85%, and PBP2a detection 98 and 100%. All methods are very good for detecting MRSA, choosing a method to use will depend on each laboratory infrastructure.

Targeted exome sequencing and chromosomal microarray for the molecular diagnosis of nevoid basal cell carcinoma syndrome.

PubMed

Matsudate, Yoshihiro; Naruto, Takuya; Hayashi, Yumiko; Minami, Mitsuyoshi; Tohyama, Mikiko; Yokota, Kenji; Yamada, Daisuke; Imoto, Issei; Kubo, Yoshiaki

2017-06-01

Nevoid basal cell carcinoma syndrome (NBCCS) is an autosomal dominant disorder mainly caused by heterozygous mutations of PTCH1. In addition to characteristic clinical features, detection of a mutation in causative genes is reliable for the diagnosis of NBCCS; however, no mutations have been identified in some patients using conventional methods. To improve the method for the molecular diagnosis of NBCCS. We performed targeted exome sequencing (TES) analysis using a multi-gene panel, including PTCH1, PTCH2, SUFU, and other sonic hedgehog signaling pathway-related genes, based on next-generation sequencing (NGS) technology in 8 cases in whom possible causative mutations were not detected by previously performed conventional analysis and 2 recent cases of NBCCS. Subsequent analysis of gross deletion within or around PTCH1 detected by TES was performed using chromosomal microarray (CMA). Through TES analysis, specific single nucleotide variants or small indels of PTCH1 causing inferred amino acid changes were identified in 2 novel cases and 2 undiagnosed cases, whereas gross deletions within or around PTCH1, which are validated by CMA, were found in 3 undiagnosed cases. However, no mutations were detected even by TES in 3 cases. Among 3 cases with gross deletions of PTCH1, deletions containing the entire PTCH1 and additional neighboring genes were detected in 2 cases, one of which exhibited atypical clinical features, such as severe mental retardation, likely associated with genes located within the 4.3Mb deleted region, especially. TES-based simultaneous evaluation of sequences and copy number status in all targeted coding exons by NGS is likely to be more useful for the molecular diagnosis of NBCCS than conventional methods. CMA is recommended as a subsequent analysis for validation and detailed mapping of deleted regions, which may explain the atypical clinical features of NBCCS cases. Copyright © 2017 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.

Detection of a Diverse Marine Fish Fauna Using Environmental DNA from Seawater Samples

PubMed Central

Iversen, Lars Lønsmann; Møller, Peter Rask; Rasmussen, Morten; Willerslev, Eske

2012-01-01

Marine ecosystems worldwide are under threat with many fish species and populations suffering from human over-exploitation. This is greatly impacting global biodiversity, economy and human health. Intriguingly, marine fish are largely surveyed using selective and invasive methods, which are mostly limited to commercial species, and restricted to particular areas with favourable conditions. Furthermore, misidentification of species represents a major problem. Here, we investigate the potential of using metabarcoding of environmental DNA (eDNA) obtained directly from seawater samples to account for marine fish biodiversity. This eDNA approach has recently been used successfully in freshwater environments, but never in marine settings. We isolate eDNA from ½-litre seawater samples collected in a temperate marine ecosystem in Denmark. Using next-generation DNA sequencing of PCR amplicons, we obtain eDNA from 15 different fish species, including both important consumption species, as well as species rarely or never recorded by conventional monitoring. We also detect eDNA from a rare vagrant species in the area; European pilchard (Sardina pilchardus). Additionally, we detect four bird species. Records in national databases confirmed the occurrence of all detected species. To investigate the efficiency of the eDNA approach, we compared its performance with 9 methods conventionally used in marine fish surveys. Promisingly, eDNA covered the fish diversity better than or equal to any of the applied conventional methods. Our study demonstrates that even small samples of seawater contain eDNA from a wide range of local fish species. Finally, in order to examine the potential dispersal of eDNA in oceans, we performed an experiment addressing eDNA degradation in seawater, which shows that even small (100-bp) eDNA fragments degrades beyond detectability within days. Although further studies are needed to validate the eDNA approach in varying environmental conditions, our findings provide a strong proof-of-concept with great perspectives for future monitoring of marine biodiversity and resources. PMID:22952584

Species-Specific Detection and Identification of Fusarium Species Complex, the Causal Agent of Sugarcane Pokkah Boeng in China

PubMed Central

Que, Youxiong; Wang, Jihua; Comstock, Jack C.; Wei, Jinjin; McCord, Per H.; Chen, Baoshan; Chen, Rukai; Zhang, Muqing

2014-01-01

Background Pokkah boeng disease caused by the Fusarium species complex results in significant yield losses in sugarcane. Thus, the rapid and accurate detection and identification of the pathogen is urgently required to manage and prevent the spreading of sugarcane pokkah boeng. Methods A total of 101 isolates were recovered from the pokkah boeng samples collected from five major sugarcane production areas in China throughout 2012 and 2013. The causal pathogen was identified by morphological observation, pathogenicity test, and phylogenetic analysis based on the fungus-conserved rDNA-ITS. Species-specific TaqMan real-time PCR and conventional PCR methods were developed for rapid and accurate detection of the causal agent of sugarcane pokkah boeng. The specificity and sensitivity of PCR assay were also evaluated on a total of 84 isolates of Fusarium from China and several isolates from other fungal pathogens of Sporisorium scitamineum and Phoma sp. and sugarcane endophyte of Acremonium sp. Result Two Fusarium species (F. verticillioides and F. proliferatum) that caused sugarcane pokahh boeng were identified by morphological observation, pathogenicity test, and phylogenetic analysis. Species-specific TaqMan PCR and conventional PCR were designed and optimized to target their rDNA-ITS regions. The sensitivity of the TaqMan PCR was approximately 10 pg of fungal DNA input, which was 1,000-fold over conventional PCR, and successfully detected pokkah boeng in the field-grown sugarcane. Conclusions/Significance This study was the first to identify two species, F. verticillioides and F. proliferatum, that were causal pathogens of sugarcane pokkah boeng in China. It also described the development of a species-specific PCR assay to detect and confirm these pathogens in sugarcane plants from mainland China. This method will be very useful for a broad range of research endeavors as well as the regulatory response and management of sugarcane pokkah boeng. PMID:25141192

Detection of a diverse marine fish fauna using environmental DNA from seawater samples.

PubMed

Thomsen, Philip Francis; Kielgast, Jos; Iversen, Lars Lønsmann; Møller, Peter Rask; Rasmussen, Morten; Willerslev, Eske

2012-01-01

Marine ecosystems worldwide are under threat with many fish species and populations suffering from human over-exploitation. This is greatly impacting global biodiversity, economy and human health. Intriguingly, marine fish are largely surveyed using selective and invasive methods, which are mostly limited to commercial species, and restricted to particular areas with favourable conditions. Furthermore, misidentification of species represents a major problem. Here, we investigate the potential of using metabarcoding of environmental DNA (eDNA) obtained directly from seawater samples to account for marine fish biodiversity. This eDNA approach has recently been used successfully in freshwater environments, but never in marine settings. We isolate eDNA from ½-litre seawater samples collected in a temperate marine ecosystem in Denmark. Using next-generation DNA sequencing of PCR amplicons, we obtain eDNA from 15 different fish species, including both important consumption species, as well as species rarely or never recorded by conventional monitoring. We also detect eDNA from a rare vagrant species in the area; European pilchard (Sardina pilchardus). Additionally, we detect four bird species. Records in national databases confirmed the occurrence of all detected species. To investigate the efficiency of the eDNA approach, we compared its performance with 9 methods conventionally used in marine fish surveys. Promisingly, eDNA covered the fish diversity better than or equal to any of the applied conventional methods. Our study demonstrates that even small samples of seawater contain eDNA from a wide range of local fish species. Finally, in order to examine the potential dispersal of eDNA in oceans, we performed an experiment addressing eDNA degradation in seawater, which shows that even small (100-bp) eDNA fragments degrades beyond detectability within days. Although further studies are needed to validate the eDNA approach in varying environmental conditions, our findings provide a strong proof-of-concept with great perspectives for future monitoring of marine biodiversity and resources.

Evaluation of Two Types of Differential Item Functioning in Factor Mixture Models with Binary Outcomes

ERIC Educational Resources Information Center

Lee, HwaYoung; Beretvas, S. Natasha

2014-01-01

Conventional differential item functioning (DIF) detection methods (e.g., the Mantel-Haenszel test) can be used to detect DIF only across observed groups, such as gender or ethnicity. However, research has found that DIF is not typically fully explained by an observed variable. True sources of DIF may include unobserved, latent variables, such as…

A Single-Step Enrichment Medium for Nonchromogenic Isolation of Healthy and Cold-Injured Salmonella spp. from Fresh Vegetables.

PubMed

Kim, Hong-Seok; Choi, Dasom; Kang, Il-Byeong; Kim, Dong-Hyeon; Yim, Jin-Hyeok; Kim, Young-Ji; Chon, Jung-Whan; Oh, Deog-Hwan; Seo, Kun-Ho

2017-02-01

Culture-based detection of nontyphoidal Salmonella spp. in foods requires at least four working days; therefore, new detection methods that shorten the test time are needed. In this study, we developed a novel single-step Salmonella enrichment broth, SSE-1, and compared its detection capability with that of commercial single-step ONE broth-Salmonella (OBS) medium and a conventional two-step enrichment method using buffered peptone water and Rappaport-Vassiliadis soy broth (BPW-RVS). Minimally processed lettuce samples were artificially inoculated with low levels of healthy and cold-injured Salmonella Enteritidis (10 0 or 10 1 colony-forming unit/25 g), incubated in OBS, BPW-RVS, and SSE-1 broths, and streaked on xylose lysine deoxycholate (XLD) agar. Salmonella recoverability was significantly higher in BPW-RVS (79.2%) and SSE-1 (83.3%) compared to OBS (39.3%) (p < 0.05). Our data suggest that the SSE-1 single-step enrichment broth could completely replace two-step enrichment with reduced enrichment time from 48 to 24 h, performing better than commercial single-step enrichment medium in the conventional nonchromogenic Salmonella detection, thus saving time, labor, and cost.

Human rhinovirus C infections in pediatric hematology and oncology patients.

PubMed

Loria, Carolina; Domm, Jennifer A; Halasa, Natasha B; Heitman, Elizabeth; Miller, E Kathryn; Xu, Meng; Saville, Benjamin R; Frangoul, Haydar; Williams, John V

2015-02-01

Children with cancer and HSCT recipients are at high risk for common viral infections. We sought to define the viral etiology of ARI and identify risk factors. Nasal wash samples were collected from pediatric hematology-oncology patients and HSCT recipients with ARI during the 2003-2005 winter seasons. Real-time RT-PCR was performed to detect Flu A, influenza B, RSV, PIV 1-3, human MPV, and HRV. HRV specimens were sequenced and genotyped. Seventy-eight samples from 62 children were included. Viruses were detected in 31 of 78 samples (40%). HRV were detected most frequently, in 16 (52%) including five HRVC; followed by seven (22%) RSV, five (16%) Flu A, four (13%) MPV, and two (6%) PIV2. There was a trend toward higher risk of viral infection for children in day care. Only 8% of the study children had received influenza vaccine. HRV, including the recently discovered HRVC, are an important cause of infection in pediatric oncology and HSCT patients. Molecular testing is superior to conventional methods and should be standard of care, as HRV are not detected by conventional methods. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Comparative performance of two quantitative safety signalling methods: implications for use in a pharmacovigilance department.

PubMed

Almenoff, June S; LaCroix, Karol K; Yuen, Nancy A; Fram, David; DuMouchel, William

2006-01-01

There is increasing interest in using disproportionality-based signal detection methods to support postmarketing safety surveillance activities. Two commonly used methods, empirical Bayes multi-item gamma Poisson shrinker (MGPS) and proportional reporting ratio (PRR), perform differently with respect to the number and types of signals detected. The goal of this study was to compare and analyse the performance characteristics of these two methods, to understand why they differ and to consider the practical implications of these differences for a large, industry-based pharmacovigilance department. We compared the numbers and types of signals of disproportionate reporting (SDRs) obtained with MGPS and PRR using two postmarketing safety databases and a simulated database. We recorded signal counts and performed a qualitative comparison of the drug-event combinations signalled by the two methods as well as a sensitivity analysis to better understand how the thresholds commonly used for these methods impact their performance. PRR detected more SDRs than MGPS. We observed that MGPS is less subject to confounding by demographic factors because it employs stratification and is more stable than PRR when report counts are low. Simulation experiments performed using published empirical thresholds demonstrated that PRR detected false-positive signals at a rate of 1.1%, while MGPS did not detect any statistical false positives. In an attempt to separate the effect of choice of signal threshold from more fundamental methodological differences, we performed a series of experiments in which we modified the conventional threshold values for each method so that each method detected the same number of SDRs for the example drugs studied. This analysis, which provided quantitative examples of the relationship between the published thresholds for the two methods, demonstrates that the signalling criterion published for PRR has a higher signalling frequency than that published for MGPS. The performance differences between the PRR and MGPS methods are related to (i) greater confounding by demographic factors with PRR; (ii) a higher tendency of PRR to detect false-positive signals when the number of reports is small; and (iii) the conventional thresholds that have been adapted for each method. PRR tends to be more 'sensitive' and less 'specific' than MGPS. A high-specificity disproportionality method, when used in conjunction with medical triage and investigation of critical medical events, may provide an efficient and robust approach to applying quantitative methods in routine postmarketing pharmacovigilance.

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.

A comprehensive evaluation of the accuracy of cervical pre-cancer detection methods in a high-risk area in East Congo

PubMed Central

Hovland, S; Arbyn, M; Lie, A K; Ryd, W; Borge, B; Berle, E J; Skomedal, H; Kadima, T M; Kyembwa, L; Billay, E M; Mukwege, D; Chirimwami, R B; Mvula, T M; Snijders, P J; Meijer, C J L M; Karlsen, F

2010-01-01

Background: Given the high burden of cervical cancer in low-income settings, there is a need for a convenient and affordable method for detecting and treating pre-cancerous lesions. Methods: Samples for comparing the accuracy of cytology, virology and histology were collected. Identification of HPV E6/E7 mRNA was performed using PreTect HPV-Proofer. HPV DNA detection was performed by GP5+/6+ PCR, followed by reverse line blot (RLB) for typing. Results: A total of 343 women, aged 25–60 years, attending gynaecological polyclinics in DR Congo were included for sample enrolment. The test positivity rate was conventional and liquid-based cytology (LBC) at cutoff ASCUS+ of 6.9 and 6.6%, respectively; PreTect HPV-Proofer of 7.3% and consensus DNA PCR for 14 HR types of 18.5%. Sixteen cases of CIN2+ lesions were identified. Of these, conventional cytology identified 66.7% with a specificity of 96.2%, LBC identified 73.3% with a specificity of 96.9%, all at cutoff ASCUS+. HR-HPV DNA detected all CIN2+ cases with a specificity of 85.9%, whereas PreTect HPV-Proofer gave a sensitivity of 81.3% and a specificity of 96.6%. Conclusion: Both HPV detection assays showed a higher sensitivity for CIN2+ than did cytological methods. Detecting E6/E7 mRNA from only a subset of HR HPVs, as is the case with PreTect HPV-Proofer, resulted in a similar specificity to cytology and a significantly higher specificity than consensus HR HPV DNA (P<0.0001). PMID:20197765

Dental enamel defect diagnosis through different technology-based devices.

PubMed

Kobayashi, Tatiana Yuriko; Vitor, Luciana Lourenço Ribeiro; Carrara, Cleide Felício Carvalho; Silva, Thiago Cruvinel; Rios, Daniela; Machado, Maria Aparecida Andrade Moreira; Oliveira, Thais Marchini

2018-06-01

Dental enamel defects (DEDs) are faulty or deficient enamel formations of primary and permanent teeth. Changes during tooth development result in hypoplasia (a quantitative defect) and/or hypomineralisation (a qualitative defect). To compare technology-based diagnostic methods for detecting DEDs. Two-hundred and nine dental surfaces of anterior permanent teeth were selected in patients, 6-11 years of age, with cleft lip with/without cleft palate. First, a conventional clinical examination was conducted according to the modified Developmental Defects of Enamel Index (DDE Index). Dental surfaces were evaluated using an operating microscope and a fluorescence-based device. Interexaminer reproducibility was determined using the kappa test. To compare groups, McNemar's test was used. Cramer's V test was used for comparing the distribution of index codes obtained after classification of all dental surfaces. Cramer's V test revealed statistically significant differences (P < .0001) in the distribution of index codes obtained using the different methods; the coefficients were 0.365 for conventional clinical examination versus fluorescence, 0.961 for conventional clinical examination versus operating microscope and 0.358 for operating microscope versus fluorescence. The sensitivity of the operating microscope and fluorescence method was statistically significant (P = .008 and P < .0001, respectively). Otherwise, the results did not show statistically significant differences in accuracy and specificity for either the operating microscope or the fluorescence methods. This study suggests that the operating microscope performed better than the fluorescence-based device and could be an auxiliary method for the detection of DEDs. © 2017 FDI World Dental Federation.

Near field ice detection using infrared based optical imaging technology

NASA Astrophysics Data System (ADS)

Abdel-Moati, Hazem; Morris, Jonathan; Zeng, Yousheng; Corie, Martin Wesley; Yanni, Victor Garas

2018-02-01

If not detected and characterized, icebergs can potentially pose a hazard to oil and gas exploration, development and production operations in arctic environments as well as commercial shipping channels. In general, very large bergs are tracked and predicted using models or satellite imagery. Small and medium bergs are detectable using conventional marine radar. As icebergs decay they shed bergy bits and growlers, which are much smaller and more difficult to detect. Their low profile above the water surface, in addition to occasional relatively high seas, makes them invisible to conventional marine radar. Visual inspection is the most common method used to detect bergy bits and growlers, but the effectiveness of visual inspections is reduced by operator fatigue and low light conditions. The potential hazard from bergy bits and growlers is further increased by short detection range (<1 km). As such, there is a need for robust and autonomous near-field detection of such smaller icebergs. This paper presents a review of iceberg detection technology and explores applications for infrared imagers in the field. Preliminary experiments are performed and recommendations are made for future work, including a proposed imager design which would be suited for near field ice detection.

Development of SCAR markers and UP-PCR cross-hybridization method for specific detection of four major subgroups of Rhizoctonia from infected turfgrasses

USDA-ARS?s Scientific Manuscript database

Several species and hyphal anastomosis groups (AG) of Rhizoctonia solani (sensu lato) cause brown patch diseases of turfgrasses. Conventional methods of identification of Rhizoctonia pathogens are time consuming and often inaccurate. A rapid identification assay for Waitea circinata (anamorph: Rhizo...

Multi-laboratory evaluations of the performance of Catellicoccus marimammalium PCR assays developed to target gull fecal sources

EPA Science Inventory

Here we report results from a multi-laboratory (n=11) evaluation of four different PCR methods targeting the 16S rRNA gene of Catellicoccus marimammalium used to detect fecal contamination from birds in coastal environments. The methods included conventional end-point PCR, a SYBR...

Optical Production and Detection of Ultrasonic Waves in Metals for Nondestructive Testing

NASA Technical Reports Server (NTRS)

Morrison, R. A.

1972-01-01

Ultrasonic waves were produced by striking the surface of a metal with the focused one-joule pulse of a Q-switched ruby laser. Rayleigh (surface) waves and longitudinal waves were detected with conventional transducers. Optical methods of detection were tested and developed. Rayleigh waves were produced with an oscillator and transducer. They were optically detected on curved polished surfaces, and on unpolished surfaces. The technique uses a knife edge to detect small angle changes of the surface as the wave pulse passes the illuminated spot. Optical flaw detection using pulse echo and attenuation is demonstrated.

Principles and applications of polymerase chain reaction in medical diagnostic fields: a review

PubMed Central

Valones, Marcela Agne Alves; Guimarães, Rafael Lima; Brandão, Lucas André Cavalcanti; de Souza, Paulo Roberto Eleutério; de Albuquerque Tavares Carvalho, Alessandra; Crovela, Sergio

2009-01-01

Recent developments in molecular methods have revolutionized the detection and characterization of microorganisms in a broad range of medical diagnostic fields, including virology, mycology, parasitology, microbiology and dentistry. Among these methods, Polymerase Chain Reaction (PCR) has generated great benefits and allowed scientific advancements. PCR is an excellent technique for the rapid detection of pathogens, including those difficult to culture. Along with conventional PCR techniques, Real-Time PCR has emerged as a technological innovation and is playing an ever-increasing role in clinical diagnostics and research laboratories. Due to its capacity to generate both qualitative and quantitative results, Real-Time PCR is considered a fast and accurate platform. The aim of the present literature review is to explore the clinical usefulness and potential of both conventional PCR and Real-Time PCR assays in diverse medical fields, addressing its main uses and advances. PMID:24031310

microRNA biosensors: Opportunities and challenges among conventional and commercially available techniques.

PubMed

Kilic, Tugba; Erdem, Arzum; Ozsoz, Mehmet; Carrara, Sandro

2018-01-15

As being the most extensively studied, non-coding, evolutionary conserved, post-transcriptional gene regulators of genome, microRNAs (miRNAs) have taken great attention among various disciplines due to their important roles in biological processes and link with cancer. Due to their diagnostic value, there have been many conventional methods used in detection of miRNAs including northern blotting, quantitative real time PCR (qRT-PCR) and microarray technology besides novel techniques based on various nanotechnology approaches and molecular biology tools including miRNA biosensors. The aim of this review is to explain the importance of miRNAs in biomedical field with an emphasis on early cancer diagnosis by overviewing both research based and commercially available miRNA detection methods in the last decade considering their strengths and weakness with an emphasis on miRNA biosensors. Copyright © 2017 Elsevier B.V. All rights reserved.

Motion-compensated detection of heart rate based on the time registration adaptive filter

NASA Astrophysics Data System (ADS)

Yang, Lei; Zhou, Jinsong; Jing, Juanjuan; Li, Yacan; Wei, Lidong; Feng, Lei; He, Xiaoying; Bu, Meixia; Fu, Xilu

2018-01-01

A non-contact heart rate detection method based on the dual-wavelength technique is proposed and demonstrated experimentally. The heart rate is obtained based on the PhotoPlethysmoGraphy (PPG). Each detection module uses the reflection detection probe which is composed of the LED and the photodiode. It is a well-known fact that the differences in the circuits of two detection modules result in different responses of two modules for motion artifacts. It will cause a time delay between the two signals. This poses a great challenge to compensate the motion artifacts during measurements. In order to solve this problem, we have firstly used the time registration and translated the signals to ensure that the two signals are consistent in time domain. Then the adaptive filter is used to compensate the motion artifacts. Moreover, the data obtained by using this non-contact detection system is compared with those of the conventional finger blood volume pulse (BVP) sensor by simultaneously measuring the heart rate of the subject. During the experiment, the left hand remains stationary and is detected by a conventional finger BVP sensor. Meanwhile, the moving palm of right hand is detected by the proposed system. The data obtained from the proposed non-contact system are consistent and comparable with that of the BVP sensor. This method can effectively suppress the interference caused by the two circuit differences and successfully compensate the motion artifacts. This technology can be used in medical and daily heart rate measurement.

Comparative study on conventional, ultrasonication and microwave assisted extraction of γ-oryzanol from rice bran.

PubMed

Kumar, Pramod; Yadav, Devbrat; Kumar, Pradyuman; Panesar, Paramjeet Singh; Bunkar, Durga Shankar; Mishra, Diwaker; Chopra, H K

2016-04-01

In present study, conventional, ultrasonic and microwave assisted extraction methods were compared with the aim of optimizing best fitting solvent and method, solvent concentration and digestion time for high yield of γ-oryzanol from rice bran. Petroleum ether, hexane and methanol were used to prepare extracts. Extraction yield were evaluated for giving high crude oil yield, total phenolic content (TPC) and γ-oryzanol content. Gas chromatography-mass spectrophotometry was used for the determination of γ-oryzanol concentration. The highest concentration of γ-oryzanol was detected in methanolic extracts of microwave treatment (85.0 ppm) followed by ultrasonication (82.0 ppm) and conventional extraction method (73.5 ppm). Concentration of γ-oryzanol present in the extracts was found to be directly proportional to the total phenolic content. A combination of 80 % methanolic concentration and 55 minutes digestion time of microwave treatment yielded the best extraction method for TPC and thus γ-oryzanol (105 ppm).

Beyond Blood Culture and Gram Stain Analysis: A Review of Molecular Techniques for the Early Detection of Bacteremia in Surgical Patients

PubMed Central

Kaplan, Heidi B.; Dua, Anahita; Litwin, Douglas B.; Ambrose, Catherine G.; Moore, Laura J.; Murray, COL Clinton K.; Wade, Charles E.; Holcomb, John B.

2016-01-01

Abstract Background: Sepsis from bacteremia occurs in 250,000 cases annually in the United States, has a mortality rate as high as 60%, and is associated with a poorer prognosis than localized infection. Because of these high figures, empiric antibiotic administration for patients with systemic inflammatory response syndrome (SIRS) and suspected infection is the second most common indication for antibiotic administration in intensive care units (ICU)s. However, overuse of empiric antibiotics contributes to the development of opportunistic infections, antibiotic resistance, and the increase in multi-drug-resistant bacterial strains. The current method of diagnosing and ruling out bacteremia is via blood culture (BC) and Gram stain (GS) analysis. Methods: Conventional and molecular methods for diagnosing bacteremia were reviewed and compared. The clinical implications, use, and current clinical trials of polymerase chain reaction (PCR)-based methods to detect bacterial pathogens in the blood stream were detailed. Results: BC/GS has several disadvantages. These include: some bacteria do not grow in culture media; others do not GS appropriately; and cultures can require up to 5 d to guide or discontinue antibiotic treatment. PCR-based methods can be potentially applied to detect rapidly, accurately, and directly microbes in human blood samples. Conclusions: Compared with the conventional BC/GS, particular advantages to molecular methods (specifically, PCR-based methods) include faster results, leading to possible improved antibiotic stewardship when bacteremia is not present. PMID:26918696

Hierarchical Flowerlike Gold Nanoparticles Labeled Immunochromatography Test Strip for Highly Sensitive Detection of Escherichia coli O157:H7.

PubMed

Zhang, Lei; Huang, Youju; Wang, Jingyun; Rong, Yun; Lai, Weihua; Zhang, Jiawei; Chen, Tao

2015-05-19

Gold nanoparticles (AuNPs) labeled lateral-flow test strip immunoassay (LFTS) has been widely used in biomedical, feed/food, and environmental analysis fields. Conventional ILFS assay usually uses spherical AuNPs as labeled probes and shows low detection sensitivity, which further limits its widespread practical application. Unlike spherical AuNP used as labeled probe in conventional ILFS, in our present study, a hierarchical flowerlike AuNP specific probe was designed for LFTS and further used to detect Escherichia coli O157:H7 (E. coli O157:H7). Three types of hierarchical flowerlike AuNPs, such as tipped flowerlike, popcornlike, and large-sized flowerlike AuNPs were synthesized in a one-step method. Compared with other two kinds of Au particles, tipped flowerlike AuNPs probes for LFTS particularly exhibited highly sensitive detection of E. coli O157:H7. The remarkable improvement of detection sensitivity of tipped flowerlike AuNPs probes can be achieved even as low as 10(3) colony-forming units (CFU)/mL by taking advantages of its appropriate size and hierarchical structures, which is superior over the detection performance of conventional LFTS. Using this novel tipped flower AuNPs probes, quantitative detection of E. coli O157:H7 can be obtained partially in a wide concentration range with good repeatability. This hierarchical tipped flower-shaped AuNPs probe for LFTS is promising for the practical applications in widespread analysis fields.

Improving label-free detection of circulating melanoma cells by photoacoustic flow cytometry

NASA Astrophysics Data System (ADS)

Zhou, Huan; Wang, Qiyan; Pang, Kai; Zhou, Quanyu; Yang, Ping; He, Hao; Wei, Xunbin

2018-02-01

Melanoma is a kind of a malignant tumor of melanocytes with the properties of high mortality and high metastasis rate. The circulating melanoma cells with the high content of melanin can be detected by light absorption to diagnose and treat cancer at an early stage. Compared with conventional detection methods such as in vivo flow cytometry (IVFC) based on fluorescence, the in vivo photoacoustic flow cytometry (PAFC) utilizes melanin cells as biomarkers to collect the photoacoustic (PA) signals without toxic fluorescent dyes labeling in a non-invasive way. The information of target tumor cells is helpful for data analysis and cell counting. However, the raw signals in PAFC system contain numerous noises such as environmental noise, device noise and in vivo motion noise. Conventional denoising algorithms such as wavelet denoising (WD) method and means filter (MF) method are based on the local information to extract the data of clinical interest, which remove the subtle feature and leave many noises. To address the above questions, the nonlocal means (NLM) method based on nonlocal data has been proposed to suppress the noise in PA signals. Extensive experiments on in vivo PA signals from the mice with the injection of B16F10 cells in caudal vein have been conducted. All the results indicate that the NLM method has superior noise reduction performance and subtle information reservation.

Left atrium by echocardiography in clinical practice: from conventional methods to new echocardiographic techniques.

PubMed

Ancona, Roberta; Comenale Pinto, Salvatore; Caso, Pio; D'Andrea, Antonello; Di Salvo, Giovanni; Arenga, Fortunato; Coppola, Maria Gabriella; Sellitto, Vincenzo; Macrino, Maria; Calabrò, Raffaele

2014-01-01

Although often referred to as "the forgotten chamber", compared with left ventricle (LV), especially in the past years, the left atrium (LA) plays a critical role in the clinical expression and prognosis of patients with heart and cerebrovascular disease, as demonstrated by several studies. Echocardiographers initially focused on early detection of atrial geometrical abnormalities through monodimensional atrial diameter quantification and then bidimensional (2D) areas and volume estimation. Now, together with conventional echocardiographic parameters, new echocardiographic techniques, such as strain Doppler, 2D speckle tracking and three-dimensional (3D) echocardiography, allow assessing early LA dysfunction and they all play a fundamental role to detect early functional remodelling before anatomical alterations occur. LA dysfunction and its important prognostic implications may be detected sooner by LA strain than by volumetric measurements.

Three-dimensional fluorescent microscopy via simultaneous illumination and detection at multiple planes.

PubMed

Ma, Qian; Khademhosseinieh, Bahar; Huang, Eric; Qian, Haoliang; Bakowski, Malina A; Troemel, Emily R; Liu, Zhaowei

2016-08-16

The conventional optical microscope is an inherently two-dimensional (2D) imaging tool. The objective lens, eyepiece and image sensor are all designed to capture light emitted from a 2D 'object plane'. Existing technologies, such as confocal or light sheet fluorescence microscopy have to utilize mechanical scanning, a time-multiplexing process, to capture a 3D image. In this paper, we present a 3D optical microscopy method based upon simultaneously illuminating and detecting multiple focal planes. This is implemented by adding two diffractive optical elements to modify the illumination and detection optics. We demonstrate that the image quality of this technique is comparable to conventional light sheet fluorescent microscopy with the advantage of the simultaneous imaging of multiple axial planes and reduced number of scans required to image the whole sample volume.

Rapid Diagnosis of Bloodstream Infections with PCR Followed by Mass Spectrometry

PubMed Central

Jordana-Lluch, Elena; Carolan, Heather E.; Giménez, Montserrat; Sampath, Rangarajan; Ecker, David J.; Quesada, M. Dolores; Mòdol, Josep M.; Arméstar, Fernando; Blyn, Lawrence B.; Cummins, Lendell L.; Ausina, Vicente; Martró, Elisa

2013-01-01

Achieving a rapid microbiological diagnosis is crucial for decreasing morbidity and mortality of patients with a bloodstream infection, as it leads to the administration of an appropriate empiric antimicrobial therapy. Molecular methods may offer a rapid alternative to conventional microbiological diagnosis involving blood culture. In this study, the performance of a new technology that uses broad-spectrum PCR coupled with mass spectrometry (PCR/ESI-MS) was evaluated for the detection of microorganisms directly from whole blood. A total of 247 whole blood samples and paired blood cultures were prospectively obtained from 175 patients with a suspicion of sepsis. Both sample types were analyzed using the PCR/ESI-MS technology, and the results were compared with those obtained by conventional identification methods. The overall agreement between conventional methods and PCR/ESI-MS performed in blood culture aliquots was 94.2% with 96.8% sensitivity and 98.5% specificity for the molecular method. When comparing conventional methods with PCR/ESI-MS performed in whole blood specimens, the overall agreement was 77.1% with 50% sensitivity and 93.8% specificity for the molecular method. Interestingly, the PCR/ESI-MS technology led to the additional identification of 13 pathogens that were not found by conventional methods. Using the PCR/ESI-MS technology the microbiological diagnosis of bloodstream infections could be anticipated in about half of the patients in our setting, including a small but significant proportion of patients newly diagnosed. Thus, this promising technology could be very useful for the rapid diagnosis of sepsis in combination with traditional methods. PMID:23626775

Plasmonic Enzyme-Linked Immunosorbent Assay Using Nanospherical Brushes as a Catalase Container for Colorimetric Detection of Ultralow Concentrations of Listeria monocytogenes.

PubMed

Chen, Rui; Huang, Xiaolin; Xu, Hengyi; Xiong, Yonghua; Li, Yanbin

2015-12-30

Plasmonic enzyme-linked immunosorbent assay (pELISA) based on catalase (CAT)-mediated gold nanoparticle growth exhibits ultrahigh sensitivity for detecting disease-related biomarkers using sandwich formats. However, the limit of detection (LOD) of this strategy for Listeria monocytogenes is only around 10(3) CFU/mL, which considerably exceeds the amount of L. monocytogenes commonly present in food products (<100 CFU/g). Herein, we report an improved pELISA method for detection of L. monocytogenes at ultralow concentrations with the sandwich formats using silica nanoparticles carrying poly(acrylic acid) brushes as a "CAT container" to increase enzyme loading for enhancing the detection signal. Under optimal conditions, the proposed pELISA exhibits good specificity and excellent sensitivity for L. monocytogenes with a LOD of 8 × 10(1) CFU/mL in 0.01 M phosphate-buffered saline, via a reaction that can be discriminated by the naked eye. The LOD obtained by this method was 2 and 5 orders of magnitude lower than that of conventional CAT-based pELISA and horseradish peroxidase (HRP)-based conventional ELISA, respectively. Coupled with large-volume immunomagnetic separation, the LOD for L. monocytogenes-spiked lettuce samples reached 8 × 10(1) CFU/g. The improved pELISA also exhibited a great potential in detecting a single cell of L. monocytogenes in 100 μL of solution.

Quantitative T1 and T2* carotid atherosclerotic plaque imaging using a three-dimensional multi-echo phase-sensitive inversion recovery sequence: a feasibility study.

PubMed

Fujiwara, Yasuhiro; Maruyama, Hirotoshi; Toyomaru, Kanako; Nishizaka, Yuri; Fukamatsu, Masahiro

2018-06-01

Magnetic resonance imaging (MRI) is widely used to detect carotid atherosclerotic plaques. Although it is important to evaluate vulnerable carotid plaques containing lipids and intra-plaque hemorrhages (IPHs) using T 1 -weighted images, the image contrast changes depending on the imaging settings. Moreover, to distinguish between a thrombus and a hemorrhage, it is useful to evaluate the iron content of the plaque using both T 1 -weighted and T 2 *-weighted images. Therefore, a quantitative evaluation of carotid atherosclerotic plaques using T 1 and T 2 * values may be necessary for the accurate evaluation of plaque components. The purpose of this study was to determine whether the multi-echo phase-sensitive inversion recovery (mPSIR) sequence can improve T 1 contrast while simultaneously providing accurate T 1 and T 2 * values of an IPH. T 1 and T 2 * values measured using mPSIR were compared to values from conventional methods in phantom and in vivo studies. In the phantom study, the T 1 and T 2 * values estimated using mPSIR were linearly correlated with those of conventional methods. In the in vivo study, mPSIR demonstrated higher T 1 contrast between the IPH phantom and sternocleidomastoid muscle than the conventional method. Moreover, the T 1 and T 2 * values of the blood vessel wall and sternocleidomastoid muscle estimated using mPSIR were correlated with values measured by conventional methods and with values reported previously. The mPSIR sequence improved T 1 contrast while simultaneously providing accurate T 1 and T 2 * values of the neck region. Although further study is required to evaluate the clinical utility, mPSIR may improve carotid atherosclerotic plaque detection and provide detailed information about plaque components.

Comparison of a real-time PCR method with a culture method for the detection of Salmonella enterica serotype enteritidis in naturally contaminated environmental samples from integrated poultry houses.

PubMed

Lungu, Bwalya; Waltman, W Douglas; Berghaus, Roy D; Hofacre, Charles L

2012-04-01

Conventional culture methods have traditionally been considered the "gold standard" for the isolation and identification of foodborne bacterial pathogens. However, culture methods are labor-intensive and time-consuming. A Salmonella enterica serotype Enteritidis-specific real-time PCR assay that recently received interim approval by the National Poultry Improvement Plan for the detection of Salmonella Enteritidis was evaluated against a culture method that had also received interim National Poultry Improvement Plan approval for the analysis of environmental samples from integrated poultry houses. The method was validated with 422 field samples collected by either the boot sock or drag swab method. The samples were cultured by selective enrichment in tetrathionate broth followed by transfer onto a modified semisolid Rappaport-Vassiliadis medium and then plating onto brilliant green with novobiocin and xylose lysine brilliant Tergitol 4 plates. One-milliliter aliquots of the selective enrichment broths from each sample were collected for DNA extraction by the commercial PrepSEQ nucleic acid extraction assay and analysis by the Salmonella Enteritidis-specific real-time PCR assay. The real-time PCR assay detected no significant differences between the boot sock and drag swab samples. In contrast, the culture method detected a significantly higher number of positive samples from boot socks. The diagnostic sensitivity of the real-time PCR assay for the field samples was significantly higher than that of the culture method. The kappa value obtained was 0.46, indicating moderate agreement between the real-time PCR assay and the culture method. In addition, the real-time PCR method had a turnaround time of 2 days compared with 4 to 8 days for the culture method. The higher sensitivity as well as the reduction in time and labor makes this real-time PCR assay an excellent alternative to conventional culture methods for diagnostic purposes, surveillance, and research studies to improve food safety.

Quantitative PCR analysis reveals a high incidence of large intragenic deletions in the FANCA gene in Spanish Fanconi anemia patients.

PubMed

Callén, E; Tischkowitz, M D; Creus, A; Marcos, R; Bueren, J A; Casado, J A; Mathew, C G; Surrallés, J

2004-01-01

Fanconi anaemia is an autosomal recessive disease characterized by chromosome fragility, multiple congenital abnormalities, progressive bone marrow failure and a high predisposition to develop malignancies. Most of the Fanconi anaemia patients belong to complementation group FA-A due to mutations in the FANCA gene. This gene contains 43 exons along a 4.3-kb coding sequence with a very heterogeneous mutational spectrum that makes the mutation screening of FANCA a difficult task. In addition, as the FANCA gene is rich in Alu sequences, it was reported that Alu-mediated recombination led to large intragenic deletions that cannot be detected in heterozygous state by conventional PCR, SSCP analysis, or DNA sequencing. To overcome this problem, a method based on quantitative fluorescent multiplex PCR was proposed to detect intragenic deletions in FANCA involving the most frequently deleted exons (exons 5, 11, 17, 21 and 31). Here we apply the proposed method to detect intragenic deletions in 25 Spanish FA-A patients previously assigned to complementation group FA-A by FANCA cDNA retroviral transduction. A total of eight heterozygous deletions involving from one to more than 26 exons were detected. Thus, one third of the patients carried a large intragenic deletion that would have not been detected by conventional methods. These results are in agreement with previously published data and indicate that large intragenic deletions are one of the most frequent mutations leading to Fanconi anaemia. Consequently, this technology should be applied in future studies on FANCA to improve the mutation detection rate. Copyright 2003 S. Karger AG, Basel

Superposition of polarized waves at layered media: theoretical modeling and measurement

NASA Astrophysics Data System (ADS)

Finkele, Rolf; Wanielik, Gerd

1997-12-01

The detection of ice layers on road surfaces is a crucial requirement for a system that is designed to warn vehicle drivers of hazardous road conditions. In the millimeter wave regime at 76 GHz the dielectric constant of ice and conventional road surface materials (i.e. asphalt, concrete) is found to be nearly similar. Thus, if the layer of ice is very thin and thus is of the same shape of roughness as the underlying road surface it cannot be securely detected using conventional algorithmic approaches. The method introduced in this paper extents and applies the theoretical work of Pancharatnam on the superposition of polarized waves. The projection of the Stokes vectors onto the Poincare sphere traces a circle due to the variation of the thickness of the ice layer. The paper presents a method that utilizes the concept of wave superposition to detect this trace even if it is corrupted by stochastic variation due to rough surface scattering. Measurement results taken under real traffic conditions prove the validity of the proposed algorithms. Classification results are presented and the results discussed.

Mover Position Detection for PMTLM Based on Linear Hall Sensors through EKF Processing

PubMed Central

Yan, Leyang; Zhang, Hui; Ye, Peiqing

2017-01-01

Accurate mover position is vital for a permanent magnet tubular linear motor (PMTLM) control system. In this paper, two linear Hall sensors are utilized to detect the mover position. However, Hall sensor signals contain third-order harmonics, creating errors in mover position detection. To filter out the third-order harmonics, a signal processing method based on the extended Kalman filter (EKF) is presented. The limitation of conventional processing method is first analyzed, and then EKF is adopted to detect the mover position. In the EKF model, the amplitude of the fundamental component and the percentage of the harmonic component are taken as state variables, and they can be estimated based solely on the measured sensor signals. Then, the harmonic component can be calculated and eliminated. The proposed method has the advantages of faster convergence, better stability and higher accuracy. Finally, experimental results validate the effectiveness and superiority of the proposed method. PMID:28383505

Real-time traffic sign recognition based on a general purpose GPU and deep-learning

PubMed Central

Hong, Yongwon; Choi, Yeongwoo; Byun, Hyeran

2017-01-01

We present a General Purpose Graphics Processing Unit (GPGPU) based real-time traffic sign detection and recognition method that is robust against illumination changes. There have been many approaches to traffic sign recognition in various research fields; however, previous approaches faced several limitations when under low illumination or wide variance of light conditions. To overcome these drawbacks and improve processing speeds, we propose a method that 1) is robust against illumination changes, 2) uses GPGPU-based real-time traffic sign detection, and 3) performs region detecting and recognition using a hierarchical model. This method produces stable results in low illumination environments. Both detection and hierarchical recognition are performed in real-time, and the proposed method achieves 0.97 F1-score on our collective dataset, which uses the Vienna convention traffic rules (Germany and South Korea). PMID:28264011

Micrometastases in neuroblastoma: are they clinically important?

PubMed Central

Burchill, S A

2004-01-01

Despite advances in the treatment of neuroblastoma (NBL), recurrence and metastases continue to pose major problems in clinical management. The relation between micrometastases and the development of secondary disease is not fully understood. However, accurate methods to detect low numbers of tumour cells may allow the evaluation of their role in the disease process, and by implication the possible benefits of eliminating them. Although there is substantial evidence for the increased sensitivity of current molecular methods for the detection of NBL cells compared with more conventional cytology, the clinical relevance and usefulness of detecting this disease remain controversial. The primary goal of current translational research must be to evaluate the clinical relevance of micrometastatic disease detected by these methods in multicentre prospective clinical outcome studies. Only then can the clinical usefulness of these methods be defined so that they may be introduced into relevant clinical practice. PMID:14693828

Optical Detection of Ultrasound in Photoacoustic Imaging

PubMed Central

Dong, Biqin; Sun, Cheng; Zhang, Hao F.

2017-01-01

Objective Photoacoustic (PA) imaging emerges as a unique tool to study biological samples based on optical absorption contrast. In PA imaging, piezoelectric transducers are commonly used to detect laser-induced ultrasonic waves. However, they typically lack adequate broadband sensitivity at ultrasonic frequency higher than 100 MHz while their bulky size and optically opaque nature cause technical difficulties in integrating PA imaging with conventional optical imaging modalities. To overcome these limitations, optical methods of ultrasound detection were developed and shown their unique applications in photoacoustic imaging. Methods We provide an overview of recent technological advances in optical methods of ultrasound detection and their applications in PA imaging. A general theoretical framework describing sensitivity, bandwidth, and angular responses of optical ultrasound detection is also introduced. Results Optical methods of ultrasound detection can provide improved detection angle and sensitivity over significantly extended bandwidth. In addition, its versatile variants also offer additional advantages, such as device miniaturization, optical transparency, mechanical flexibility, minimal electrical/mechanical crosstalk, and potential noncontact PA imaging. Conclusion The optical ultrasound detection methods discussed in this review and their future evolution may play an important role in photoacoustic imaging for biomedical study and clinical diagnosis. PMID:27608445

Stability Analysis of Radial Turning Process for Superalloys

NASA Astrophysics Data System (ADS)

Jiménez, Alberto; Boto, Fernando; Irigoien, Itziar; Sierra, Basilio; Suarez, Alfredo

2017-09-01

Stability detection in machining processes is an essential component for the design of efficient machining processes. Automatic methods are able to determine when instability is happening and prevent possible machine failures. In this work a variety of methods are proposed for detecting stability anomalies based on the measured forces in the radial turning process of superalloys. Two different methods are proposed to determine instabilities. Each one is tested on real data obtained in the machining of Waspalloy, Haynes 282 and Inconel 718. Experimental data, in both Conventional and High Pressure Coolant (HPC) environments, are set in four different states depending on materials grain size and Hardness (LGA, LGS, SGA and SGS). Results reveal that PCA method is useful for visualization of the process and detection of anomalies in online processes.

Detection of Legionella species in environmental water by the quantitative PCR method in combination with ethidium monoazide treatment.

PubMed

Inoue, Hiroaki; Takama, Tomoko; Yoshizaki, Miwa; Agata, Kunio

2015-01-01

We detected Legionella species in 111 bath water samples and 95 cooling tower water samples by using a combination of conventional plate culture, quantitative polymerase chain reaction (qPCR) and qPCR combined with ethidium monoazide treatment (EMA-qPCR) methods. In the case of bath water samples, Legionella spp. were detected in 30 samples by plate culture, in 85 samples by qPCR, and in 49 samples by EMA-qPCR. Of 81 samples determined to be Legionella-negative by plate culture, 56 and 23 samples were positive by qPCR and EMA-qPCR, respectively. Therefore, EMA treatment decreased the number of Legionella-positive bath water samples detected by qPCR. In contrast, EMA treatment had no effect on cooling tower water samples. We therefore expect that EMA-qPCR is a useful method for the rapid detection of viable Legionella spp. from bath water samples.

Detection of Pseudomonas savastanoi pv. savastanoi in olive plants by enrichment and PCR.

PubMed

Penyalver, R; García, A; Ferrer, A; Bertolini, E; López, M M

2000-06-01

The sequence of the gene iaaL of Pseudomonas savastanoi EW2009 was used to design primers for PCR amplification. The iaaL-derived primers directed the amplification of a 454-bp fragment from genomic DNA isolated from 70 strains of P. savastanoi, whereas genomic DNA from 93 non-P. savastanoi isolates did not yield this amplified product. A previous bacterial enrichment in the semiselective liquid medium PVF-1 improved the PCR sensitivity level, allowing detection of 10 to 100 CFU/ml of plant extract. P. savastanoi was detected by the developed enrichment-PCR method in knots from different varieties of inoculated and naturally infected olive trees. Moreover, P. savastanoi was detected in symptomless stem tissues from naturally infected olive plants. This enrichment-PCR method is more sensitive and less cumbersome than the conventional isolation methods for detection of P. savastanoi.

Detection of the Dinozoans Pfiesteria piscicida and P. shumwayae: a review of detection methods and geographic distribution.

PubMed

Rublee, Parke A; Remington, David L; Schaefer, Eric F; Marshall, Michael M

2005-01-01

Molecular methods, including conventional PCR, real-time PCR, denaturing gradient gel electrophoresis, fluorescent fragment detection PCR, and fluorescent in situ hybridization, have all been developed for use in identifying and studying the distribution of the toxic dinoflagellates Pfiesteria piscicida and P. shumwayae. Application of the methods has demonstrated a worldwide distribution of both species and provided insight into their environmental tolerance range and temporal changes in distribution. Genetic variability among geographic locations generally appears low in rDNA genes, and detection of the organisms in ballast water is consistent with rapid dispersal or high gene flow among populations, but additional sequence data are needed to verify this hypothesis. The rapid development and application of these tools serves as a model for study of other microbial taxa and provides a basis for future development of tools that can simultaneously detect multiple targets.

Detection of heavy metal by paper-based microfluidics.

PubMed

Lin, Yang; Gritsenko, Dmitry; Feng, Shaolong; Teh, Yi Chen; Lu, Xiaonan; Xu, Jie

2016-09-15

Heavy metal pollution has shown great threat to the environment and public health worldwide. Current methods for the detection of heavy metals require expensive instrumentation and laborious operation, which can only be accomplished in centralized laboratories. Various microfluidic paper-based analytical devices have been developed recently as simple, cheap and disposable alternatives to conventional ones for on-site detection of heavy metals. In this review, we first summarize current development of paper-based analytical devices and discuss the selection of paper substrates, methods of device fabrication, and relevant theories in these devices. We then compare and categorize recent reports on detection of heavy metals using paper-based microfluidic devices on the basis of various detection mechanisms, such as colorimetric, fluorescent, and electrochemical methods. To finalize, the future development and trend in this field are discussed. Copyright © 2016 Elsevier B.V. All rights reserved.

Demonstration of a Speckle Based Sensing with Pulse-Doppler Radar for Vibration Detection.

PubMed

Ozana, Nisan; Bauer, Reuven; Ashkenazy, Koby; Sasson, Nissim; Schwarz, Ariel; Shemer, Amir; Zalevsky, Zeev

2018-05-03

In previous works, an optical technique for extraction and separation of remote static vibrations has been demonstrated. In this paper, we will describe an approach in which RF speckle movement is used to extract remote vibrations of a static target. The use of conventional radar Doppler methods is not suitable for detecting vibrations of static targets. In addition, the speckle method has an important advantage, in that it is able to detect vibrations at far greater distances than what is normally detected in classical optical methods. The experiment described in this paper was done using a motorized vehicle, which engine was turned on and off. The results showed that the system was able to distinguish between the different engine states, and in addition, was able to determine the vibration frequency of the engine. The first step towards real time detection of human vital signs using RF speckle patterns is presented.

Detection of 22 common leukemic fusion genes using a single-step multiplex qRT-PCR-based assay.

PubMed

Lyu, Xiaodong; Wang, Xianwei; Zhang, Lina; Chen, Zhenzhu; Zhao, Yu; Hu, Jieying; Fan, Ruihua; Song, Yongping

2017-07-25

Fusion genes generated from chromosomal translocation play an important role in hematological malignancies. Detection of fusion genes currently employ use of either conventional RT-PCR methods or fluorescent in situ hybridization (FISH), where both methods involve tedious methodologies and require prior characterization of chromosomal translocation events as determined by cytogenetic analysis. In this study, we describe a real-time quantitative reverse transcription PCR (qRT-PCR)-based multi-fusion gene screening method with the capacity to detect 22 fusion genes commonly found in leukemia. This method does not require pre-characterization of gene translocation events, thereby facilitating immediate diagnosis and therapeutic management. We performed fluorescent qRT-PCR (F-qRT-PCR) using a commercially-available multi-fusion gene detection kit on a patient cohort of 345 individuals comprising 108 cases diagnosed with acute myeloid leukemia (AML) for initial evaluation; remaining patients within the cohort were assayed for confirmatory diagnosis. Results obtained by F-qRT-PCR were compared alongside patient analysis by cytogenetic characterization. Gene translocations detected by F-qRT-PCR in AML cases were diagnosed in 69.4% of the patient cohort, which was comparatively similar to 68.5% as diagnosed by cytogenetic analysis, thereby demonstrating 99.1% concordance. Overall gene fusion was detected in 53.7% of the overall patient population by F-qRT-PCR, 52.9% by cytogenetic prediction in leukemia, and 9.1% in non-leukemia patients by both methods. The overall concordance rate was calculated to be 99.0%. Fusion genes were detected by F-qRT-PCR in 97.3% of patients with CML, followed by 69.4% with AML, 33.3% with acute lymphoblastic leukemia (ALL), 9.1% with myelodysplastic syndromes (MDS), and 0% with chronic lymphocytic leukemia (CLL). We describe the use of a F-qRT-PCR-based multi-fusion gene screening method as an efficient one-step diagnostic procedure as an effective alternative to lengthy conventional diagnostic procedures requiring both cytogenetic analysis followed by targeted quantitative reverse transcription (qRT-PCR) methods, thus allowing timely patient management.

Evaluation of permeable fractures in rock aquifers

NASA Astrophysics Data System (ADS)

Bok Lee, Hang

2015-04-01

In this study, the practical usefulness and fundamental applicability of a self-potential (SP) method for identifying the permeable fractures were evaluated by a comparison of SP methods with other geophysical logging methods and hydraulic tests. At a 10 m-shallow borehole in the study site, the candidates of permeable fractures crossing the borehole were first determined by conventional geophysical methods such as an acoustic borehole televiwer, temperature, electrical conductivity and gamma-gamma loggings, which was compared to the analysis by the SP method. Constant pressure injection and recovery tests were conducted for verification of the hydraulic properties of the fractures identified by various logging methods. The acoustic borehole televiwer and gamma-gamma loggings detected the open space or weathering zone within the borehole, but they cannot prove the possibility of a groundwater flow through the detected fractures. The temperature and electrical conductivity loggings had limitations to detect the fractured zones where groundwater in the borehole flows out to the surrounding rock aquifers. Comparison of results from different methods showed that there is a best correlation between the distribution of hydraulic conductivity and the variation of the SP signals, and the SP logging can estimate accurately the hydraulic activity as well as the location of permeable fractures. Based on the results, the SP method is recommended for determining the hydraulically-active fractures rather than other conventional geophysical loggings. This self-potential method can be effectively applied in the initial stage of a site investigation which selects the optimal location and evaluates the hydrogeological property of fractures in target sites for the underground structure including the geothermal reservoir and radioactive waste disposal.

Metamorphopsia Score and Central Visual Field Outcomes in Diabetic Cystoid Macular Edema

PubMed Central

Brzozowska, Agnieszka; Maciejewski, Ryszard

2018-01-01

Aim To detect abnormality of the visual function in naïve patients with cystoid diabetic macular edema (DME) using M-charts, Amsler test, and white on white (W/W) and blue on yellow (B/Y) perimetry. Methods There were 64 eyes included in the study: 30 eyes with DME, 22 eyes with diabetes without DME, and 12 eyes of normal subjects. Conventional W/W perimetry and B/Y perimetry were performed within the central 10° of the visual field. To assess metamorphopsia, Amsler test and M-charts were used. Results The rate of detection of metamorphopsia was 37% with Amsler test examination and 50% with M-charts. Specificity of both tests was 100%. We found a significant difference between vertical scores of M-charts in all groups, but not in horizontal scores (p < 0.0001). Mean defect (MD) was 8.9 dB and 3.6 dB and loss variance (LV) 4.8 dB and 3.3 dB (p < 0.0001). Conclusions M-chart is more sensitive than Amsler test method for detection of metamorphopsia. The MD and LV are higher in b/y in comparison to W/W perimetry. B/Y perimetry and M-charts are more sensitive than conventional methods for detecting the visual function loss in cystoid DME. PMID:29744359

Peak tree: a new tool for multiscale hierarchical representation and peak detection of mass spectrometry data.

PubMed

Zhang, Peng; Li, Houqiang; Wang, Honghui; Wong, Stephen T C; Zhou, Xiaobo

2011-01-01

Peak detection is one of the most important steps in mass spectrometry (MS) analysis. However, the detection result is greatly affected by severe spectrum variations. Unfortunately, most current peak detection methods are neither flexible enough to revise false detection results nor robust enough to resist spectrum variations. To improve flexibility, we introduce peak tree to represent the peak information in MS spectra. Each tree node is a peak judgment on a range of scales, and each tree decomposition, as a set of nodes, is a candidate peak detection result. To improve robustness, we combine peak detection and common peak alignment into a closed-loop framework, which finds the optimal decomposition via both peak intensity and common peak information. The common peak information is derived and loopily refined from the density clustering of the latest peak detection result. Finally, we present an improved ant colony optimization biomarker selection method to build a whole MS analysis system. Experiment shows that our peak detection method can better resist spectrum variations and provide higher sensitivity and lower false detection rates than conventional methods. The benefits from our peak-tree-based system for MS disease analysis are also proved on real SELDI data.

All-optical photoacoustic microscopy (AOPAM) system for remote characterization of biological tissues

NASA Astrophysics Data System (ADS)

Sampathkumar, Ashwin; Chitnis, Parag V.; Silverman, Ronald H.

2014-03-01

Conventional photoacoustic microscopy (PAM) employs light pulses to produce a photoacoustic (PA) effect and detects the resulting acoustic waves using an ultrasound transducer acoustically coupled to the target. The resolution of conventional PAM is limited by the sensitivity and bandwidth of the ultrasound transducer. We investigated a versatile, all-optical PAM (AOPAM) system for characterizing in vivo as well as ex vivo biological specimens. The system employs non-contact interferometric detection of PA signals that overcomes limitations of conventional PAM. A 532-nm pump laser with a pulse duration of 5 ns excites the PA effect in tissue. Resulting acoustic waves produce surface displacements that are sensed using a 532-nm continuous-wave (CW) probe laser in a Michelson interferometer with a 1- GHz bandwidth. The pump and probe beams are coaxially focused using a 50X objective giving a diffraction-limited spot size of 0.48 μm. The phase-encoded probe beam is demodulated using homodyne methods. The detected timedomain signal is time reversed using k-space wave-propagation methods to produce a spatial distribution of PA sources in the target tissue. A minimum surface-displacement sensitivity of 0.19 pm was measured. PA-induced surface displacements are very small; therefore, they impose stringent detection requirements and determine the feasibility of implementing an all-optical PAM in biomedical applications. 3D PA images of ex vivo porcine retina specimens were generated successfully. We believe the AOPAM system potentially is well suited for assessing retinal diseases and other near-surface biomedical applications such as sectionless histology and evaluation of skin burns and pressure or friction ulcers.

A Simple Method Based on the Application of a CCD Camera as a Sensor to Detect Low Concentrations of Barium Sulfate in Suspension

PubMed Central

de Sena, Rodrigo Caciano; Soares, Matheus; Pereira, Maria Luiza Oliveira; da Silva, Rogério Cruz Domingues; do Rosário, Francisca Ferreira; da Silva, Joao Francisco Cajaiba

2011-01-01

The development of a simple, rapid and low cost method based on video image analysis and aimed at the detection of low concentrations of precipitated barium sulfate is described. The proposed system is basically composed of a webcam with a CCD sensor and a conventional dichroic lamp. For this purpose, software for processing and analyzing the digital images based on the RGB (Red, Green and Blue) color system was developed. The proposed method had shown very good repeatability and linearity and also presented higher sensitivity than the standard turbidimetric method. The developed method is presented as a simple alternative for future applications in the study of precipitations of inorganic salts and also for detecting the crystallization of organic compounds. PMID:22346607

Development of loop-mediated isothermal amplification methods for detecting Taylorella equigenitalis and Taylorella asinigenitalis

PubMed Central

KINOSHITA, Yuta; NIWA, Hidekazu; KATAYAMA, Yoshinari; HARIU, Kazuhisa

2015-01-01

ABSTRACT Taylorella equigenitalis is a causative bacterium of contagious equine metritis (CEM), and Taylorella asinigenitalis is species belonging to genus Taylorella. The authors developed two loop-mediated isothermal amplification (LAMP) methods, Te-LAMP and Ta-LAMP, for detecting T. equigenitalis and T. asinigenitalis, respectively. Using experimentally spiked samples, Te-LAMP was as sensitive as a published semi-nested PCR method, and Ta-LAMP was more sensitive than conventional PCR. Multiplex LAMP worked well without nonspecific reactions, and the analytical sensitivities of multiplex LAMP in the spiked samples were almost equivalent to those of Te-LAMP and Ta-LAMP. Therefore, the LAMP methods are considered useful tools to detect T. equigenitalis and/or T. asinigenitalis, and preventive measures will be rapidly implemented if the occurrence of CEM is confirmed by the LAMP methods. PMID:25829868

Analysis of microdialysate monoamines, including noradrenaline, dopamine and serotonin, using capillary ultra-high performance liquid chromatography and electrochemical detection.

PubMed

Ferry, Barbara; Gifu, Elena-Patricia; Sandu, Ioana; Denoroy, Luc; Parrot, Sandrine

2014-03-01

Electrochemical methods are very often used to detect catecholamine and indolamine neurotransmitters separated by conventional reverse-phase high performance liquid chromatography (HPLC). The present paper presents the development of a chromatographic method to detect monoamines present in low-volume brain dialysis samples using a capillary column filled with sub-2μm particles. Several parameters (repeatability, linearity, accuracy, limit of detection) for this new ultrahigh performance liquid chromatography (UHPLC) method with electrochemical detection were examined after optimization of the analytical conditions. Noradrenaline, adrenaline, serotonin, dopamine and its metabolite 3-methoxytyramine were separated in 1μL of injected sample volume; they were detected above concentrations of 0.5-1nmol/L, with 2.1-9.5% accuracy and intra-assay repeatability equal to or less than 6%. The final method was applied to very low volume dialysates from rat brain containing monoamine traces. The study demonstrates that capillary UHPLC with electrochemical detection is suitable for monitoring dialysate monoamines collected at high sampling rate. Copyright © 2014 Elsevier B.V. All rights reserved.

Time-resolved methods in biophysics. 7. Photon counting vs. analog time-resolved singlet oxygen phosphorescence detection.

PubMed

Jiménez-Banzo, Ana; Ragàs, Xavier; Kapusta, Peter; Nonell, Santi

2008-09-01

Two recent advances in optoelectronics, namely novel near-IR sensitive photomultipliers and inexpensive yet powerful diode-pumped solid-state lasers working at kHz repetition rate, enable the time-resolved detection of singlet oxygen (O2(a1Deltag)) phosphorescence in photon counting mode, thereby boosting the time-resolution, sensitivity, and dynamic range of this well-established detection technique. Principles underlying this novel approach and selected examples of applications are provided in this perspective, which illustrate the advantages over the conventional analog detection mode.

Detection of urinary leakage after radical retropubic prostatectomy by contrast enhanced ultrasound - do we still need conventional retrograde cystography?

PubMed

Schoeppler, Gita M; Buchner, Alexander; Zaak, Dirk; Khoder, Wael; Staehler, Michael; Stief, Christian G; Reiser, Maximilian F; Clevert, Dirk-Andre

2010-12-01

To prospectively evaluate the accuracy of transvesical contrast-enhanced ultrasound (CEUS) as an alternative method for the detection of anastomotic leakage after radical retropubic prostatectomy (RRP) in comparison with the current standard method of conventional retrograde cystography (CG). Forty-three patients underwent RRP for histologically proven localized prostate cancer. The vesico-urethral anastomosis was evaluated 8 days after RRP by CG and CEUS. Any peri-anastomotic leakage was assessed and determined in CG and CEUS as follows: no extravasation (EV), small leakage (≤0.5 cm), moderate leakage (>0.5 cm to ≤2 cm), large leakage (>2 cm diameter of EV seen). In total, 21 (49%) patients showed a watertight anastomosis. Ten (23%), two (4.7%) and ten (23%) patients showed a small, intermediate and large EV, respectively. In 31 cases (72%) there was 100% agreement of CG and CEUS for detection of no, moderate and large EV, respectively. In nine cases a small and in two cases a moderate EV was categorized as watertight anastomosis by CEUS. Only in one case did CG detect a small EV where a large EV was detected in CEUS. The agreement between both methods was 95% for detecting absence or large leakages. CEUS is a promising imaging modality that seems to be equivalent to CG for detecting the presence of a large anastomotic leakage that is clinically relevant for postoperative persistence of the indwelling catheter. CEUS could be a cheap and time-saving alternative to the CG without exposure of the patient to radiation. © 2010 THE AUTHORS. JOURNAL COMPILATION © 2010 BJU INTERNATIONAL.

Nonlinear multi-photon laser wave-mixing optical detection in microarrays and microchips for ultrasensitive detection and separation of biomarkers for cancer and neurodegenerative diseases

NASA Astrophysics Data System (ADS)

Iwabuchi, Manna; Hetu, Marcel; Maxwell, Eric; Pradel, Jean S.; Ramos, Sashary; Tong, William G.

2015-09-01

Multi-photon degenerate four-wave mixing is demonstrated as an ultrasensitive absorption-based optical method for detection, separation and identification of biomarker proteins in the development of early diagnostic methods for HIV- 1, cancer and neurodegenerative diseases using compact, portable microarrays and capillary- or microchip-based chemical separation systems that offer high chemical specificity levels. The wave-mixing signal has a quadratic dependence on concentration, and hence, it allows more reliable monitoring of smaller changes in analyte properties. Our wave-mixing detection sensitivity is comparable or better than those of current methods including enzyme-linked immunoassay for clinical diagnostic and screening. Detection sensitivity is excellent since the wave-mixing signal is a coherent laser-like beam that can be collected with virtually 100% collection efficiency with high S/N. Our analysis time is short (1-15 minutes) for molecular weight-based protein separation as compared to that of a conventional separation technique, e.g., sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When ultrasensitive wavemixing detection is paired with high-resolution capillary- or microchip-based separation systems, biomarkers can be separated and identified at the zepto- and yocto-mole levels for a wide range of analytes. Specific analytes can be captured in a microchannel through the use of antibody-antigen interactions that provide better chemical specificity as compared to size-based separation alone. The technique can also be combined with immune-precipitation and a multichannel capillary array for high-throughput analysis of more complex protein samples. Wave mixing allows the use of chromophores and absorption-modifying tags, in addition to conventional fluorophores, for online detection of immunecomplexes related to cancer.

Evaluation of volatile organic emissions from hazardous waste incinerators.

PubMed Central

Sedman, R M; Esparza, J R

1991-01-01

Conventional methods of risk assessment typically employed to evaluate the impact of hazardous waste incinerators on public health must rely on somewhat speculative emissions estimates or on complicated and expensive sampling and analytical methods. The limited amount of toxicological information concerning many of the compounds detected in stack emissions also complicates the evaluation of the public health impacts of these facilities. An alternative approach aimed at evaluating the public health impacts associated with volatile organic stack emissions is presented that relies on a screening criterion to evaluate total stack hydrocarbon emissions. If the concentration of hydrocarbons in ambient air is below the screening criterion, volatile emissions from the incinerator are judged not to pose a significant threat to public health. Both the screening criterion and a conventional method of risk assessment were employed to evaluate the emissions from 20 incinerators. Use of the screening criterion always yielded a substantially greater estimate of risk than that derived by the conventional method. Since the use of the screening criterion always yielded estimates of risk that were greater than that determined by conventional methods and measuring total hydrocarbon emissions is a relatively simple analytical procedure, the use of the screening criterion would appear to facilitate the evaluation of operating hazardous waste incinerators. PMID:1954928

A modified Elek test for detection of toxigenic corynebacteria in the diagnostic laboratory.

PubMed

Engler, K H; Glushkevich, T; Mazurova, I K; George, R C; Efstratiou, A

1997-02-01

The detection of toxigenicity among Corynebacterium diphtheriae and Corynebacterium ulcerans strains is the most important test for the microbiological diagnosis of diphtheria. Difficulties with current methods, in particular the Elek test, are well documented. We therefore describe a modified Elek test which provides an accurate result after only 16 h of incubation, in contrast to 48 h for the conventional test.

Detection of melamine in milk powders using Near-Infrared Hyperspectral imaging combined with regression coefficient of partial least square regression model

USDA-ARS?s Scientific Manuscript database

Illegal use of nitrogen-rich melamine (C3H6N6) to boost perceived protein content of food products such as milk, infant formula, frozen yogurt, pet food, biscuits, and coffee drinks has caused serious food safety problems. Conventional methods to detect melamine in foods, such as Enzyme-linked immun...

Quantifying viruses and bacteria in wastewater—Results, interpretation methods, and quality control

USGS Publications Warehouse

Francy, Donna S.; Stelzer, Erin A.; Bushon, Rebecca N.; Brady, Amie M.G.; Mailot, Brian E.; Spencer, Susan K.; Borchardt, Mark A.; Elber, Ashley G.; Riddell, Kimberly R.; Gellner, Terry M.

2011-01-01

Membrane bioreactors (MBR), used for wastewater treatment in Ohio and elsewhere in the United States, have pore sizes small enough to theoretically reduce concentrations of protozoa and bacteria, but not viruses. Sampling for viruses in wastewater is seldom done and not required. Instead, the bacterial indicators Escherichia coli (E. coli) and fecal coliforms are the required microbial measures of effluents for wastewater-discharge permits. Information is needed on the effectiveness of MBRs in removing human enteric viruses from wastewaters, particularly as compared to conventional wastewater treatment before and after disinfection. A total of 73 regular and 28 quality-control (QC) samples were collected at three MBR and two conventional wastewater plants in Ohio during 23 regular and 3 QC sampling trips in 2008-10. Samples were collected at various stages in the treatment processes and analyzed for bacterial indicators E. coli, fecal coliforms, and enterococci by membrane filtration; somatic and F-specific coliphage by the single agar layer (SAL) method; adenovirus, enterovirus, norovirus GI and GII, rotavirus, and hepatitis A virus by molecular methods; and viruses by cell culture. While addressing the main objective of the study-comparing removal of viruses and bacterial indicators in MBR and conventional plants-it was realized that work was needed to identify data analysis and quantification methods for interpreting enteric virus and QC data. Therefore, methods for quantifying viruses, qualifying results, and applying QC data to interpretations are described in this report. During each regular sampling trip, samples were collected (1) before conventional or MBR treatment (post-preliminary), (2) after secondary or MBR treatment (post-secondary or post-MBR), (3) after tertiary treatment (one conventional plant only), and (4) after disinfection (post-disinfection). Glass-wool fiber filtration was used to concentrate enteric viruses from large volumes, and small volume grab samples were collected for direct-plating analyses for bacterial indicators and coliphage. After filtration, the viruses were eluted from the filter and further concentrated. The final concentrated sample volume (FCSV) was used for enteric virus analysis by use of two methods-cell culture and a molecular method, polymerase chain reaction (PCR). Quantitative PCR (qPCR) for DNA viruses and quantitative reverse-transcriptase PCR (qRT-PCR) for RNA viruses were used in this study. To support data interpretations, the assay limit of detection (ALOD) was set for each virus assay and used to determine sample reporting limits (SRLs). For qPCR and qRT-PCR the ALOD was an estimated value because it was not established according to established method detection limit procedures. The SRLs were different for each sample because effective sample volumes (the volume of the original sample that was actually used in each analysis) were different for each sample. Effective sample volumes were much less than the original sample volumes because of reductions from processing steps and (or) from when dilutions were made to minimize the effects from PCR-inhibiting substances. Codes were used to further qualify the virus data and indicate the level of uncertainty associated with each measurement. Quality-control samples were used to support data interpretations. Field and laboratory blanks for bacteria, coliphage, and enteric viruses were all below detection, indicating that it was unlikely that samples were contaminated from equipment or processing procedures. The absolute value log differences (AVLDs) between concurrent replicate pairs were calculated to identify the variability associated with each measurement. For bacterial indicators and coliphage, the AVLD results indicated that concentrations <10 colony-forming units or plaque-forming units per 100 mL can differ between replicates by as much as 1 log, whereas higher concentrations can differ by as much as 0.3 log. The AVLD results for viruses indicated that differences between replicates can be as great as 1.2 log genomic copies per liter, regardless of the concentration of virus. Relatively large differences in molecular results for viruses between replicate pairs were likely due to lack of precision for samples with small effective volumes. Concentrations of E. coli, fecal coliforms, enterococci, and somatic and F-specific coliphage in post-secondary and post-tertiary samples in conventional plants were higher than those in post-MBR samples. In post-MBR and post-secondary samples, concentrations of somatic coliphage were higher than F-specific coliphage. In post-disinfection samples from two MBR plants (the third MBR plant had operational issues) and the ultraviolet conventional plant, concentrations for all bacterial indicators and coliphage were near or below detection; from the chlorine conventional plant, concentrations in post-disinfection samples were in the single or double digits. All of the plants met the National Pollutant Discharge Elimination System required effluent limits established for fecal coliforms. Norovirus GII and hepatitis A virus were not detected in any samples, and rotavirus was detected in one sample but could not be quantified. Adenovirus was found in 100 percent, enterovirus in over one-half, and norovirus GI in about one-half of post-preliminary wastewater samples. Adenovirus and enterovirus were detected throughout the treatment processes, and norovirus GI was detected less often than the other two enteric viruses. Culturable viruses were detected in post-preliminary samples and in only two post-treatment samples from the plant with operational issues.

The advance of non-invasive detection methods in osteoarthritis

NASA Astrophysics Data System (ADS)

Dai, Jiao; Chen, Yanping

2011-06-01

Osteoarthritis (OA) is one of the most prevalent chronic diseases which badly affected the patients' living quality and economy. Detection and evaluation technology can provide basic information for early treatment. A variety of imaging methods in OA were reviewed, such as conventional X-ray, computed tomography (CT), ultrasound (US), magnetic resonance imaging (MRI) and near-infrared spectroscopy (NIRS). Among the existing imaging modalities, the spatial resolution of X-ray is extremely high; CT is a three-dimensional method, which has high density resolution; US as an evaluation method of knee OA discriminates lesions sensitively between normal cartilage and degenerative one; as a sensitive and nonionizing method, MRI is suitable for the detection of early OA, but the cost is too expensive for routine use; NIRS is a safe, low cost modality, and is also good at detecting early stage OA. In a word, each method has its own advantages, but NIRS is provided with broader application prospect, and it is likely to be used in clinical daily routine and become the golden standard for diagnostic detection.

Web-Based Versus Conventional Training for Medical Students on Infant Gross Motor Screening.

PubMed

Pusponegoro, Hardiono D; Soebadi, Amanda; Surya, Raymond

2015-12-01

Early detection of developmental abnormalities is important for early intervention. A simple screening method is needed for use by general practitioners, as is an effective and efficient training method. This study aims to evaluate the effectiveness, acceptability, and usability of Web-based training for medical students on a simple gross motor screening method in infants. Fifth-year medical students at University of Indonesia in Jakarta were randomized into two groups. A Web-based training group received online video modules, discussions, and assessments (at www.schoology.com ). A conventional training group received a 1-day live training using the same module. Both groups completed identical pre- and posttests and the User Satisfaction Questionnaire (USQ). The Web-based group also completed the System Usability Scale (SUS). The module was based on a gross motor screening method used in the World Health Organization Multicentre Growth Reference Study. There were 39 and 32 subjects in the Web-based and conventional groups, respectively. Mean pretest versus posttest scores (correct answers out of 20) were 9.05 versus 16.95 (p=0.0001) in the Web-based group and 9.31 versus 16.88 (p=0.0001) in the conventional group. Mean difference between pre- and posttest scores did not differ significantly between the Web-based and conventional groups (mean [standard deviation], 7.56 [3.252] versus 7.90 [5.170]; p=0.741]. Both training methods were acceptable based on USQ scores. Based on SUS scores, the Web-based training had good usability. Web-based training is an effective, efficient, and acceptable training method for medical students on simple infant gross motor screening and is as effective as conventional training.

Radar fall detection using principal component analysis

NASA Astrophysics Data System (ADS)

Jokanovic, Branka; Amin, Moeness; Ahmad, Fauzia; Boashash, Boualem

2016-05-01

Falls are a major cause of fatal and nonfatal injuries in people aged 65 years and older. Radar has the potential to become one of the leading technologies for fall detection, thereby enabling the elderly to live independently. Existing techniques for fall detection using radar are based on manual feature extraction and require significant parameter tuning in order to provide successful detections. In this paper, we employ principal component analysis for fall detection, wherein eigen images of observed motions are employed for classification. Using real data, we demonstrate that the PCA based technique provides performance improvement over the conventional feature extraction methods.

Defect Detectability Improvement for Conventional Friction Stir Welds

NASA Technical Reports Server (NTRS)

Hill, Chris

2013-01-01

This research was conducted to evaluate the effects of defect detectability via phased array ultrasound technology in conventional friction stir welds by comparing conventionally prepped post weld surfaces to a machined surface finish. A machined surface is hypothesized to improve defect detectability and increase material strength.

Capillary electrophoresis with laser-induced fluorescence detection: a sensitive method for monitoring extracellular concentrations of amino acids in the periaqueductal grey matter.

PubMed

Bergquist, J; Vona, M J; Stiller, C O; O'Connor, W T; Falkenberg, T; Ekman, R

1996-03-01

The use of capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) for the analysis of microdialysate samples from the periaqueductal grey matter (PAG) of freely moving rats is described. By employing 3-(4-carboxybenzoyl)-2-quinoline-carboxaldehyde (CBQCA) as a derivatization agent, we simultaneously monitored the concentrations of 8 amino acids (arginine, glutamine, valine, gamma-amino-n-butyric acid (GABA), alanine, glycine, glutamate, and aspartate), with nanomolar and subnanomolar detection limits. Two of the amino acids (GABA and glutamate) were analysed in parallel by conventional high-performance liquid chromatography (HPLC) in order to directly compare the two analytical methods. Other CE methods for analysis of microdialysate have been previously described, and this improved method offers greater sensitivity, ease of use, and the possibility to monitor several amino acids simultaneously. By using this technique together with an optimised form of microdialysis technique, the tiny sample consumption and the improved detection limits permit the detection of fast and transient transmitter changes.

ROKU: a novel method for identification of tissue-specific genes.

PubMed

Kadota, Koji; Ye, Jiazhen; Nakai, Yuji; Terada, Tohru; Shimizu, Kentaro

2006-06-12

One of the important goals of microarray research is the identification of genes whose expression is considerably higher or lower in some tissues than in others. We would like to have ways of identifying such tissue-specific genes. We describe a method, ROKU, which selects tissue-specific patterns from gene expression data for many tissues and thousands of genes. ROKU ranks genes according to their overall tissue specificity using Shannon entropy and detects tissues specific to each gene if any exist using an outlier detection method. We evaluated the capacity for the detection of various specific expression patterns using synthetic and real data. We observed that ROKU was superior to a conventional entropy-based method in its ability to rank genes according to overall tissue specificity and to detect genes whose expression pattern are specific only to objective tissues. ROKU is useful for the detection of various tissue-specific expression patterns. The framework is also directly applicable to the selection of diagnostic markers for molecular classification of multiple classes.

Establishment of a simple and quantitative immunospot assay for detecting anti-type II collagen antibody using an infrared fluorescence imaging system (IFIS).

PubMed

Ota, Shusuke; Kanazawa, Satoshi; Kobayashi, Masaaki; Otsuka, Takanobu; Okamoto, Takashi

2005-04-01

Antibodies to type II collagen (col II) have been detected in patients with rheumatoid arthritis and in animal models of collagen induced arthritis. Here, we describe a novel method to detect anti-col II antibodies using an immunospot assay with an infrared fluorescence imaging system. This method showed very high sensitivity and specificity, and was simple, with low background levels. It also showed higher reproducibility and linearity, with a dynamic range of approximately 500-fold, than the conventional immunospot assay with enhanced chemiluminescence detection. Using this method we were able to demonstrate the antibody affinity maturation process in mice immunized with col II. In these immunized mice, although cross-reactive antibodies reacting with other collagen species were detected in earlier stages of immunization, the titers of cross-reactive antibodies rapidly diminished after the antigen boost, concomitantly with the elevation of the anti-col II antibody. The method and its possible applications are discussed.

Quantitative mapping of total choline in healthy human breast using proton echo planar spectroscopic imaging (PEPSI) at 3 Tesla.

PubMed

Zhao, Chenguang; Bolan, Patrick J; Royce, Melanie; Lakkadi, Navneeth; Eberhardt, Steven; Sillerud, Laurel; Lee, Sang-Joon; Posse, Stefan

2012-11-01

To quantitatively measure tCho levels in healthy breasts using Proton-Echo-Planar-Spectroscopic-Imaging (PEPSI). The two-dimensional mapping of tCho at 3 Tesla across an entire breast slice using PEPSI and a hybrid spectral quantification method based on LCModel fitting and integration of tCho using the fitted spectrum were developed. This method was validated in 19 healthy females and compared with single voxel spectroscopy (SVS) and with PRESS prelocalized conventional Magnetic Resonance Spectroscopic Imaging (MRSI) using identical voxel size (8 cc) and similar scan times (∼7 min). A tCho peak with a signal to noise ratio larger than 2 was detected in 10 subjects using both PEPSI and SVS. The average tCho concentration in these subjects was 0.45 ± 0.2 mmol/kg using PEPSI and 0.48 ± 0.3 mmol/kg using SVS. Comparable results were obtained in two subjects using conventional MRSI. High lipid content in the spectra of nine tCho negative subjects was associated with spectral line broadening of more than 26 Hz, which made tCho detection impossible. Conventional MRSI with PRESS prelocalization in glandular tissue in two of these subjects yielded tCho concentrations comparable to PEPSI. The detection sensitivity of PEPSI is comparable to SVS and conventional PRESS-MRSI. PEPSI can be potentially used in the evaluation of tCho in breast cancer. A tCho threshold concentration value of ∼0.7 mmol/kg might be used to differentiate between cancerous and healthy (or benign) breast tissues based on this work and previous studies. Copyright © 2012 Wiley Periodicals, Inc.

Most Probable Number Rapid Viability PCR Method to Detect Viable Spores of Bacillus anthracis in Swab Samples

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

Letant, S E; Kane, S R; Murphy, G A

2008-05-30

This note presents a comparison of Most-Probable-Number Rapid Viability (MPN-RV) PCR and traditional culture methods for the quantification of Bacillus anthracis Sterne spores in macrofoam swabs generated by the Centers for Disease Control and Prevention (CDC) for a multi-center validation study aimed at testing environmental swab processing methods for recovery, detection, and quantification of viable B. anthracis spores from surfaces. Results show that spore numbers provided by the MPN RV-PCR method were in statistical agreement with the CDC conventional culture method for all three levels of spores tested (10{sup 4}, 10{sup 2}, and 10 spores) even in the presence ofmore » dirt. In addition to detecting low levels of spores in environmental conditions, the MPN RV-PCR method is specific, and compatible with automated high-throughput sample processing and analysis protocols.« less

An Effective Electrical Resonance-Based Method to Detect Delamination in Thermal Barrier Coating

NASA Astrophysics Data System (ADS)

Kim, Jong Min; Park, Jae-Ha; Lee, Ho Girl; Kim, Hak-Joon; Song, Sung-Jin; Seok, Chang-Sung; Lee, Young-Ze

2017-12-01

This research proposes a simple yet highly sensitive method based on electrical resonance of an eddy-current probe to detect delamination of thermal barrier coating (TBC). This method can directly measure the mechanical characteristics of TBC compared to conventional ultrasonic testing and infrared thermography methods. The electrical resonance-based method can detect the delamination of TBC from the metallic bond coat by shifting the electrical impedance of eddy current testing (ECT) probe coupling with degraded TBC, and, due to this shift, the resonant frequencies near the peak impedance of ECT probe revealed high sensitivity to the delamination. In order to verify the performance of the proposed method, a simple experiment is performed with degraded TBC specimens by thermal cyclic exposure. Consequently, the delamination with growth of thermally grown oxide in a TBC system is experimentally identified. Additionally, the results are in good agreement with the results obtained from ultrasonic C-scanning.

An Effective Electrical Resonance-Based Method to Detect Delamination in Thermal Barrier Coating

NASA Astrophysics Data System (ADS)

Kim, Jong Min; Park, Jae-Ha; Lee, Ho Girl; Kim, Hak-Joon; Song, Sung-Jin; Seok, Chang-Sung; Lee, Young-Ze

2018-02-01

This research proposes a simple yet highly sensitive method based on electrical resonance of an eddy-current probe to detect delamination of thermal barrier coating (TBC). This method can directly measure the mechanical characteristics of TBC compared to conventional ultrasonic testing and infrared thermography methods. The electrical resonance-based method can detect the delamination of TBC from the metallic bond coat by shifting the electrical impedance of eddy current testing (ECT) probe coupling with degraded TBC, and, due to this shift, the resonant frequencies near the peak impedance of ECT probe revealed high sensitivity to the delamination. In order to verify the performance of the proposed method, a simple experiment is performed with degraded TBC specimens by thermal cyclic exposure. Consequently, the delamination with growth of thermally grown oxide in a TBC system is experimentally identified. Additionally, the results are in good agreement with the results obtained from ultrasonic C-scanning.

High-Resolution Scintimammography: A Pilot Study

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

Rachel F. Brem; Joelle M. Schoonjans; Douglas A. Kieper

2002-07-01

This study evaluated a novel high-resolution breast-specific gamma camera (HRBGC) for the detection of suggestive breast lesions. Methods: Fifty patients (with 58 breast lesions) for whom a scintimammogram was clinically indicated were prospectively evaluated with a general-purpose gamma camera and a novel HRBGC prototype. The results of conventional and high-resolution nuclear studies were prospectively classified as negative (normal or benign) or positive (suggestive or malignant) by 2 radiologists who were unaware of the mammographic and histologic results. All of the included lesions were confirmed by pathology. Results: There were 30 benign and 28 malignant lesions. The sensitivity for detection ofmore » breast cancer was 64.3% (18/28) with the conventional camera and 78.6% (22/28) with the HRBGC. The specificity with both systems was 93.3% (28/30). For the 18 nonpalpable lesions, sensitivity was 55.5% (10/18) and 72.2% (13/18) with the general-purpose camera and the HRBGC, respectively. For lesions 1 cm, 7 of 15 were detected with the general-purpose camera and 10 of 15 with the HRBGC. Four lesions (median size, 8.5 mm) were detected only with the HRBGC and were missed by the conventional camera. Conclusion: Evaluation of indeterminate breast lesions with an HRBGC results in improved sensitivity for the detection of cancer, with greater improvement shown for nonpalpable and 1-cm lesions.« less

[Disappearance of residual disease confirmed by RT-PCR following induction chemotherapy in two hypoplastic leukemia patients with t(8;21)].

PubMed

Sawada, M; Tsurumi, H; Yamada, T; Hara, T; Oyama, M; Moriwaki, H

1999-04-01

Reverse transcriptase-polymerase chain reaction (RT-PCR) methods often detect the AML1/MTG8 fusion transcript even in acute myelogenous leukemia (AML) patients with t(8;21) who have been in long-term remission. We encountered 2 hypoplastic leukemia patients with t(8;21) who achieved cytogenetic remission with short-term conventional chemotherapy. Patient 1 was a 42-year-old woman. Chromosomal analysis detected t(8;21) (q22;q22) and PCR analysis (35 cycles PCR amplification; detection limit 1 x 10(-5) cells) detected the AML1/MTG8 fusion transcript. Complete remission was obtained with 1 course of chemotherapy consisting of low-dose cytarabine (20 mg x 14 days) and etoposide (50 mg x 14 days). After 2 courses of consolidation chemotherapy consisting of conventional-dose cytarabine and mitoxantrone, the RT-PCR findings were negative for the AML1/MTG8 fusion transcript. Patient 2 was a 67-year-old man. Cytogenetic analysis detected t(8;21) (q22;q22), and was positive for the AML1/MTG8 fusion transcript. After 2 courses of induction chemotherapy comprising low-dose cytarabine (20 mg x 14 days) and etoposide (50 mg x 14 days), and 3 courses of conventional consolidation chemotherapy, RT-PCR analysis confirmed the disappearance of the AML1/MTG8 fusion transcript.

Sensitivity of a real-time PCR method for the detection of transgenes in a mixture of transgenic and non-transgenic seeds of papaya (Carica papaya L.).

PubMed

Nageswara-Rao, Madhugiri; Kwit, Charles; Agarwal, Sujata; Patton, Mariah T; Skeen, Jordan A; Yuan, Joshua S; Manshardt, Richard M; Stewart, C Neal

2013-09-01

Genetically engineered (GE) ringspot virus-resistant papaya cultivars 'Rainbow' and 'SunUp' have been grown in Hawai'i for over 10 years. In Hawai'i, the introduction of GE papayas into regions where non-GE cultivars are grown and where feral non-GE papayas exist have been accompanied with concerns associated with transgene flow. Of particular concern is the possibility of transgenic seeds being found in non-GE papaya fruits via cross-pollination. Development of high-throughput methods to reliably detect the adventitious presence of such transgenic material would benefit both the scientific and regulatory communities. We assessed the accuracy of using conventional qualitative polymerase chain reaction (PCR) as well as real-time PCR-based assays to quantify the presence of transgenic DNA from bulk samples of non-GE papaya seeds. In this study, an optimized method of extracting high quality DNA from dry seeds of papaya was standardized. A reliable, sensitive real-time PCR method for detecting and quantifying viral coat protein (cp) transgenes in bulk seed samples utilizing the endogenous papain gene is presented. Quantification range was from 0.01 to 100 ng/μl of GE-papaya DNA template with a detection limit as low as 0.01% (10 pg). To test this system, we simulated transgene flow using known quantities of GE and non-GE DNA and determined that 0.038% (38 pg) GE papaya DNA could be detected using real-time PCR. We also validated this system by extracting DNA from known ratios of GE seeds to non-GE seeds of papaya followed by real-time PCR detection and observed a reliable detection limit of 0.4%. This method for the quick and sensitive detection of transgenes in bulked papaya seed lots using conventional as well as real-time PCR-based methods will benefit numerous stakeholders. In particular, this method could be utilized to screen selected fruits from maternal non-GE papaya trees in Hawai'i for the presence of transgenic seed at typical regulatory threshold levels. Incorporation of subtle differences in primers and probes for variations in cp worldwide should allow this method to be utilized elsewhere when and if deregulation of transgenic papaya occurs.

Sensitivity of a real-time PCR method for the detection of transgenes in a mixture of transgenic and non-transgenic seeds of papaya (Carica papaya L.)

PubMed Central

2013-01-01

Background Genetically engineered (GE) ringspot virus-resistant papaya cultivars ‘Rainbow’ and ‘SunUp’ have been grown in Hawai’i for over 10 years. In Hawai’i, the introduction of GE papayas into regions where non-GE cultivars are grown and where feral non-GE papayas exist have been accompanied with concerns associated with transgene flow. Of particular concern is the possibility of transgenic seeds being found in non-GE papaya fruits via cross-pollination. Development of high-throughput methods to reliably detect the adventitious presence of such transgenic material would benefit both the scientific and regulatory communities. Results We assessed the accuracy of using conventional qualitative polymerase chain reaction (PCR) as well as real-time PCR-based assays to quantify the presence of transgenic DNA from bulk samples of non-GE papaya seeds. In this study, an optimized method of extracting high quality DNA from dry seeds of papaya was standardized. A reliable, sensitive real-time PCR method for detecting and quantifying viral coat protein (cp) transgenes in bulk seed samples utilizing the endogenous papain gene is presented. Quantification range was from 0.01 to 100 ng/μl of GE-papaya DNA template with a detection limit as low as 0.01% (10 pg). To test this system, we simulated transgene flow using known quantities of GE and non-GE DNA and determined that 0.038% (38 pg) GE papaya DNA could be detected using real-time PCR. We also validated this system by extracting DNA from known ratios of GE seeds to non-GE seeds of papaya followed by real-time PCR detection and observed a reliable detection limit of 0.4%. Conclusions This method for the quick and sensitive detection of transgenes in bulked papaya seed lots using conventional as well as real-time PCR-based methods will benefit numerous stakeholders. In particular, this method could be utilized to screen selected fruits from maternal non-GE papaya trees in Hawai’i for the presence of transgenic seed at typical regulatory threshold levels. Incorporation of subtle differences in primers and probes for variations in cp worldwide should allow this method to be utilized elsewhere when and if deregulation of transgenic papaya occurs. PMID:24004548

Lab-on-a-chip and SDS-PAGE analysis of hemolymph protein profile from Rhipicephalus microplus (Acari: Ixodidae) infected with entomopathogenic nematode and fungus.

PubMed

Golo, Patrícia Silva; Dos Santos, Alessa Siqueira de Oliveira; Monteiro, Caio Marcio Oliveira; Perinotto, Wendell Marcelo de Souza; Quinelato, Simone; Camargo, Mariana Guedes; de Sá, Fillipe Araujo; Angelo, Isabele da Costa; Martins, Marta Fonseca; Prata, Marcia Cristina de Azevedo; Bittencourt, Vânia Rita Elias Pinheiro

2016-09-01

In the present study, lab-on-a-chip electrophoresis (LoaC) was suggested as an alternative method to the conventional polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE) to analyze raw cell-free tick hemolymph. Rhipicephalus microplus females were exposed to the entomopathogenic fungus Metarhizium anisopliae senso latu IBCB 116 strain and/or to the entomopathogenic nematode Heterorhabditis indica LPP1 strain. Hemolymph from not exposed or exposed ticks was collected 16 and 24 h after exposure and analyze by SDS-PAGE or LoaC. SDS-PAGE yielded 15 bands and LoaC electrophoresis 17 bands. Despite the differences in the number of bands, when the hemolymph protein profiles of exposed or unexposed ticks were compared in the same method, no suppressing or additional bands were detected among the treatments regardless the method (i.e., SDS-PAGE or chip electrophoresis using the Protein 230 Kit®). The potential of LoaC electrophoresis to detect protein bands from tick hemolymph was considered more efficient in comparison to the detection obtained using the traditional SDS-PAGE method, especially when it comes to protein subunits heavier than 100 KDa. LoaC electrophoresis provided a very good reproducibility, and is much faster than the conventional SDS-PAGE method, which requires several hours for one analysis. Despite both methods can be used to analyze tick hemolymph composition, LoaC was considered more suitable for cell-free hemolymph protein separation and detection. LoaC hemolymph band percent data reported changes in key proteins (i.e., HeLp and vitellogenin) exceptionally important for tick embryogenesis. This study reported, for the first time, tick hemolymph protein profile using LoaC.

Simultaneous HPLC determination of flavonoids and phenolic acids profile in Pêra-Rio orange juice.

PubMed

Mesquita, E; Monteiro, M

2018-04-01

The aim of this study was to develop and validate an HPLC-DAD method to evaluate the phenolic compounds profile of organic and conventional Pêra-Rio orange juice. The proposed method was validated for 10 flavonoids and 6 phenolic acids. A wide linear range (0.01-223.4μg·g -1 ), good accuracy (79.5-129.2%) and precision (CV≤3.8%), low limits of detection (1-22ng·g -1 ) and quantification (0.7-7.4μg), and overall ruggedness were attained. Good recovery was achieved for all phenolic compounds after extraction and cleanup. The method was applied to organic and conventional Pêra-Rio orange juices from beginning, middle and end of the 2016 harvest. Flavones rutin, nobiletin and tangeretin, and flavanones hesperidin, narirutin and eriocitrin were identified and quantified in all organic and conventional juices. Identity was confirmed by mass spectrometry. Nineteen non-identified phenolic compounds were quantified based on DAD spectra characteristic of the chemical class: 7 cinnamic acid derivatives, 6 flavanones and 6 flavones. The phenolic compounds profile of Pêra-Rio orange juices changed during the harvest; levels increased in organic orange juices, and decreased or were about the same in conventional orange juices. Phenolic compounds levels were higher in organic (0.5-1143.7mg·100g -1 ) than in conventional orange juices (0.5-689.7mg·100g -1 ). PCA differentiated organic from conventional FS and NFC juices, and conventional FCOJ from conventional FS and NFC juices, thus differentiating cultivation and processing. Copyright © 2017. Published by Elsevier Ltd.

Advances in biological dosimetry

NASA Astrophysics Data System (ADS)

Ivashkevich, A.; Ohnesorg, T.; Sparbier, C. E.; Elsaleh, H.

2017-01-01

Rapid retrospective biodosimetry methods are essential for the fast triage of persons occupationally or accidentally exposed to ionizing radiation. Identification and detection of a radiation specific molecular ‘footprint’ should provide a sensitive and reliable measurement of radiation exposure. Here we discuss conventional (cytogenetic) methods of detection and assessment of radiation exposure in comparison to emerging approaches such as gene expression signatures and DNA damage markers. Furthermore, we provide an overview of technical and logistic details such as type of sample required, time for sample preparation and analysis, ease of use and potential for a high throughput analysis.

Determination of morphine and codeine in urine using poly(dimethylsiloxane) microchip electrophoresis with electrochemical detection.

PubMed

Zhang, Qian-Li; Xu, Jing-Juan; Li, Xiang-Yun; Lian, Hong-Zhen; Chen, Hong-Yuan

2007-01-04

In this paper, a poly(dimethylsiloxane) (PDMS) microchip with electrochemical (EC) detection was developed for rapid separation and detection of morphine and codeine. It was found that morphine and codeine were well separated within 140 s in phosphate buffer solution (PBS) (pH 6.6, 40 mM)-beta-cyclodextrin (beta-CD) (20 mM)-acetonitrile (30%, v/v). The detection limit was 0.2 microM for morphine and 1 microM for codeine. The protocol was successfully applied to monitoring the amount of morphine and codeine in human urine. Compared with the conventional methods, the presented method had many advantages such as lower instrument cost, less reagent consumption and shorter analysis time.

RAMA casein zymography: Time-saving and highly sensitive casein zymography for MMP7 and trypsin.

PubMed

Yasumitsu, Hidetaro; Ozeki, Yasuhiro; Kanaly, Robert A

2016-11-01

To detect metalloproteinase-7 (MMP7), zymography is conducted using a casein substrate and conventional CBB stain. It has disadvantages because it is time consuming and has low sensitivity. Previously, a sensitive method to detect MMP7 up to 30 pg was reported, however it required special substrates and complicated handlings. RAMA casein zymography described herein is rapid, sensitive, and reproducible. By applying high-sensitivity staining with low substrate conditions, the staining process is completed within 1 h and sensitivity was increased 100-fold. The method can detect 10 pg MMP7 by using commercially available casein without complicated handlings. Moreover, it increases detection sensitivity for trypsin. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

POLLUTION PREVENTION AND ENHANCEMENT OF BIODEGRADABILITY VIA ISOMER ELIMINATION IN CONSUMER PRODUCTS

EPA Science Inventory

The purpose of this project is to develop novel methodologies for the analysis and detection of chiral environmental contaminants. Conventional analytical techniques do not discriminate between enantiomers. By using newly developed enantioselective methods, the environmental pers...

Optimizing Imaging Instruments for Emission Mammography

NASA Astrophysics Data System (ADS)

Weinberg, Irving N.

1996-05-01

Clinical studies have demonstrated that radiotracer methods can noninvasively detect breast cancers in vivo(L.P. Adler, J.P.Crowe, N.K. Al-Kaisis, et al, Radiology 187,743-750 (1993)) (I. Khalkhali, I. Mena, E. Jouanne, et al, J. Am. Coll. Surg. 178, 491-497 (1994)). Due to spatial resolution and count efficiency considerations, users of conventional nuclear medicine instruments have had difficulty in detecting subcentimeter cancers. This limitation is unfortunate, since cancer therapy is generally most efficacious when tumor diameter at detection is less than a centimeter. A more subtle limitation of conventional nuclear medicine imaging instruments is that they are poorly suited to guiding interventions. With the assistance of C.J. Thompson from McGill University, and the CEBAF Detector Physics Group, we have explored the possibility of configuring detectors for nuclear medicine imaging devices into geometries that resemble conventional x-ray mammography cameras(I.N. Weinberg, U.S.Patent 5,252,830 (1993)). Phantom and pilot clinical studies suggest that applying breast compression within such geometries may offer several advantages(C.J. Thompson, K. Murthy, I.N. Weinberg, et al, Med. Physics 21, 259-538 (1994)): For coincident detection of positron emitters, efficiency and spatial resolution are improved by bringing the detectors very close to the source (the breast tumor). For single-photon detection, attenuation due to overlying tissue is reduced. Since, for a high-efficiency collimator, spatial resolution worsens with increasing source to collimator distance, adoption of compression allows more efficient collimators to be employed. Economics are favorable in that detectors can be deployed in the region of interest, rather than around the entire body, and that such detectors can be mounted in conventional mammographic gantries. The application of conventional mammographic geometry promises to assist physicians in conducting radiotracer-guided biopsies, and in correlating biochemical with x-ray data. The primary challenge of conducting studies with dedicated emission mammography devices has been dealing with high count rates due to cardiac activity.

A Lateral Flow Biosensor for the Detection of Single Nucleotide Polymorphisms.

PubMed

Zeng, Lingwen; Xiao, Zhuo

2017-01-01

A lateral flow biosensor (LFB) is introduced for the detection of single nucleotide polymorphisms (SNPs). The assay is composed of two steps: circular strand displacement reaction and lateral flow biosensor detection. In step 1, the nucleotide at SNP site is recognized by T4 DNA ligase and the signal is amplified by strand displacement DNA polymerase, which can be accomplished at a constant temperature. In step 2, the reaction product of step 1 is detected by a lateral flow biosensor, which is a rapid and cost effective tool for nuclei acid detection. Comparing with conventional methods, it requires no complicated machines. It is suitable for the use of point of care diagnostics. Therefore, this simple, cost effective, robust, and promising LFB detection method of SNP has great potential for the detection of genetic diseases, personalized medicine, cancer related mutations, and drug-resistant mutations of infectious agents.

Invasive pulmonary aspergillosis: current diagnostic methodologies and a new molecular approach.

PubMed

Moura, S; Cerqueira, L; Almeida, A

2018-05-13

The fungus Aspergillus fumigatus is the main pathogenic agent responsible for invasive pulmonary aspergillosis. Immunocompromised patients are more likely to develop this pathology due to a decrease in the immune system's defense capacity. Despite of the low occurrence of invasive pulmonary aspergillosis, this pathology presents high rates of mortality, mostly due to late and unspecific diagnosis. Currently, the diagnostic methods used to detect this fungal infection are conventional mycological examination (direct microscopic examination, histological examination, and culture), imaging, non-culture-based tests for the detection of galactomannan, β(1,3)-glucan and an extracellular glycoprotein, and molecular tests based on PCR. However, most of these methods do not detect the species A. fumigatus; they only allow the identification of genus Aspergillus. The development of more specific detection methods is of extreme importance. Fluorescent in situ hybridization-based molecular methods can be a good alternative to achieve this purpose. In this review, it is intended to point out that most of the methods used for the diagnosis of invasive pulmonary aspergillosis do not allow to detect the fungus at the species level and that fluorescence in situ hybridization-based molecular method will be a promising approach in the A. fumigatus detection.

Rationalizing and advancing the 3-MPBA SERS sandwich assay for rapid detection of bacteria in environmental and food matrices.

PubMed

Pearson, Brooke; Mills, Alexander; Tucker, Madeline; Gao, Siyue; McLandsborough, Lynne; He, Lili

2018-06-01

Bacterial foodborne illness continues to be a pressing issue in our food supply. Rapid detection methods are needed for perishable foods due to their short shelf lives and significant contribution to foodborne illness. Previously, a sensitive and reliable surface-enhanced Raman spectroscopy (SERS) sandwich assay based on 3-mercaptophenylboronic acid (3-MBPA) as a capturer and indicator molecule was developed for rapid bacteria detection. In this study, we explored the advantages and constraints of this assay over the conventional aerobic plate count (APC) method and further developed methods for detection in real environmental and food matrices. The SERS sandwich assay was able to detect environmental bacteria in pond water and on spinach leaves at higher levels than the APC method. In addition, the SERS assay appeared to have higher sensitivity to quantify bacteria in the stationary phase. On the other hand, the APC method was more sensitive to cell viability. Finally, a method to detect bacteria in a challenging high-sugar juice matrix was developed to enhance bacteria capture. This study advanced the SERS technique for real applications in environment and food matrices. Copyright © 2017 Elsevier Ltd. All rights reserved.

The detection of T-Nos, a genetic element present in GMOs, by cross-priming isothermal amplification with real-time fluorescence.

PubMed

Zhang, Fang; Wang, Liu; Fan, Kai; Wu, Jian; Ying, Yibin

2014-05-01

An isothermal cross-priming amplification (CPA) assay for Agrobacterium tumefaciens nopaline synthase terminator (T-Nos) was established and investigated in this work. A set of six specific primers, recognizing eight distinct regions on the T-Nos sequence, was designed. The CPA assay was performed at a constant temperature, 63 °C, and detected by real-time fluorescence. The results indicated that real-time fluorescent CPA had high specificity, and the limit of detection was 1.06 × 10(3) copies of rice genomic DNA, which could be detected in 40 min. Comparison of real-time fluorescent CPA and conventional polymerase chain reaction (PCR) was also performed. Results revealed that real-time fluorescent CPA had a comparable sensitivity to conventional real-time PCR and had taken a shorter time. In addition, different contents of genetically modified (GM)-contaminated rice seed powder samples were detected for practical application. The result showed real-time fluorescent CPA could detect 0.5 % GM-contaminated samples at least, and the whole reaction could be finished in 35 min. Real-time fluorescent CPA is sensitive enough to monitor labeling systems and provides an attractive method for the detection of GMO.

groEL is a suitable genetic marker for detecting Vibrio parahaemolyticus by loop-mediated isothermal amplification assay.

PubMed

Siddique, M P; Jang, W J; Lee, J M; Ahn, S H; Suraiya, S; Kim, C H; Kong, I S

2017-08-01

A groEL gene-based loop-mediated isothermal amplification (LAMP) assay was developed to detect Vibrio parahaemolyticus in contaminated seafood and water. The assay was optimized and conducted at 63°C for 40 min using Bacillus stearothermophilus (Bst) DNA polymerase, large fragment. Amplification was analysed via multiple detection methods, including opacity, formation of white precipitate, DNA intercalating dyes (ethidium bromide and SYBR Green I), metal ion-binding indicator dye, calcein, and 2% agarose gel electrophoresis. A characteristic ladder-like band pattern on agarose gel and the desired colour changes when using different dyes were observed in positive cases, and these were species-specific for V. parahaemolyticus when compared with other closely related Vibrio spp. The limit of detection (LoD) of this assay was 100 fg per reaction, 100-fold higher than that for conventional polymerase chain reaction (PCR). When tested on artificially contaminated seafood and seawater, the LoDs of the LAMP assay were 120 and 150 fg per reaction respectively, and those of conventional PCR were 120 and 150 pg per reaction respectively. Based on our results, the groEL gene-based LAMP assay is rapid, specific, sensitive, and reliable for detecting V. parahaemolyticus, and it could be used in field diagnosis. The loop-mediated isothermal amplification (LAMP) assay using groEL gene (an abundant, highly conserved gene and member of the groESL chaperone gene family) provided rapid, species-specific and highly sensitive method for detecting Vibrio parahaemolyticus, the leading causal agent of seafood-borne diseases worldwide. Moreover, groEL LAMP revealed high efficiency than conventional PCR assay for V. parahaemolyticus using template both from pure culture and artificially contaminated seafood and water, which indicated the applicability in the field and environmental screening purpose for the organism. © 2017 The Society for Applied Microbiology.

A simple and novel modification of comet assay for determination of bacteriophage mediated bacterial cell lysis.

PubMed

Khairnar, Krishna; Sanmukh, Swapnil; Chandekar, Rajshree; Paunikar, Waman

2014-07-01

The comet assay is the widely used method for in vitro toxicity testing which is also an alternative to the use of animal models for in vivo testing. Since, its inception in 1984 by Ostling and Johansson, it is being modified frequently for a wide range of application. In spite of its wide applicability, unfortunately there is no report of its application in bacteriophages research. In this study, a novel application of comet assay for the detection of bacteriophage mediated bacterial cell lysis was described. The conventional methods in bacteriophage research for studying bacterial lysis by bacteriophages are plaque assay method. It is time consuming, laborious and costly. The lytic activity of bacteriophage devours the bacterial cell which results in the release of bacterial genomic material that gets detected by ethidium bromide staining method by the comet assay protocol. The objective of this study was to compare efficacy of comet assay with different assay used to study phage mediated bacterial lysis. The assay was performed on culture isolates (N=80 studies), modified comet assay appear to have relatively higher sensitivity and specificity than other assay. The results of the study showed that the application of comet assay can be an economical, time saving and less laborious alternative to conventional plaque assay for the detection of bacteriophage mediated bacterial cell lysis. Copyright © 2014 Elsevier B.V. All rights reserved.

Biosynthesis and computational analysis of amine-ended dual thiol ligand functionalized gold nanoparticles for conventional spectroscopy detection of melamine.

PubMed

Anand, K; Singh, Thishana; Madhumitha, G; Phulukdaree, A; Gengan, Robert M; Chuturgoon, A A

2017-04-01

The bio-synthesized DTAuNPs have an average size of 21nm. The aggregation extent depends on the concentration of melamine, which was validated by UV-vis spectra and visual method of melamine detection was developed. The major observation in this method was the color change of DTAuNPs from red to purple due to the aggregation of ligand capped gold nanoparticles instigated by melamine. The reaction of color changes were processed due to the shifting of bonding in hydrogen in between nanoparticles and melamine. The aggregation extent depends on the concentration of melamine, which can be validated UV-vis spectra and visual method of detecting melamine is developed. The electron density and conventional UV-vis, FTIR spectroscopy and DFT studies on the ligand was performed using computational methods. The theoretical and experimental data for the energy transitions and the molar extinction coefficients of the ligands studied has been obtained. Further, the ligand capped gold nanoparticles was assessed for cytotoxicity against A549 cells which resulted in significant decrease in cell viability was noted in 50μg/mL DTAu, 4-ATP and AXT treated cells at 2h (85% and 66%) and 6h (83% and 36%) respectively, (p<0.01) were studied and reported in this manuscript. Copyright © 2017 Elsevier B.V. All rights reserved.

RNase H-assisted RNA-primed rolling circle amplification for targeted RNA sequence detection.

PubMed

Takahashi, Hirokazu; Ohkawachi, Masahiko; Horio, Kyohei; Kobori, Toshiro; Aki, Tsunehiro; Matsumura, Yukihiko; Nakashimada, Yutaka; Okamura, Yoshiko

2018-05-17

RNA-primed rolling circle amplification (RPRCA) is a useful laboratory method for RNA detection; however, the detection of RNA is limited by the lack of information on 3'-terminal sequences. We uncovered that conventional RPRCA using pre-circularized probes could potentially detect the internal sequence of target RNA molecules in combination with RNase H. However, the specificity for mRNA detection was low, presumably due to non-specific hybridization of non-target RNA with the circular probe. To overcome this technical problem, we developed a method for detecting a sequence of interest in target RNA molecules via RNase H-assisted RPRCA using padlocked probes. When padlock probes are hybridized to the target RNA molecule, they are converted to the circular form by SplintR ligase. Subsequently, RNase H creates nick sites only in the hybridized RNA sequence, and single-stranded DNA is finally synthesized from the nick site by phi29 DNA polymerase. This method could specifically detect at least 10 fmol of the target RNA molecule without reverse transcription. Moreover, this method detected GFP mRNA present in 10 ng of total RNA isolated from Escherichia coli without background DNA amplification. Therefore, this method can potentially detect almost all types of RNA molecules without reverse transcription and reveal full-length sequence information.

The Detection Method of Escherichia coli in Water Resources: A Review

NASA Astrophysics Data System (ADS)

Nurliyana, M. R.; Sahdan, M. Z.; Wibowo, K. M.; Muslihati, A.; Saim, H.; Ahmad, S. A.; Sari, Y.; Mansor, Z.

2018-04-01

This article reviews several approaches for Escherichia coli (E. coli) bacteria detection from conventional methods, emerging method and goes to biosensor-based techniques. Detection and enumeration of E. coli bacteria usually required long duration of time in obtaining the result since laboratory-based approach is normally used in its assessment. It requires 24 hours to 72 hours after sampling to process the culturing samples before results are available. Although faster technique for detecting E. coli in water such as Polymerase Chain Reaction (PCR) and Enzyme-Linked Immunosorbent Assay (ELISA) have been developed, it still required transporting the samples from water resources to the laboratory, high-cost, complicated equipment usage, complex procedures, as well as the requirement of skilled specialist to cope with the complexity which limit their wide spread practice in water quality detection. Recently, development of biosensor device that is easy to perform, portable, highly sensitive and selective becomes indispensable in detecting extremely lower consolidation of pathogenic E. coli bacteria in water samples.

Comparison of real-time PCR methods for the detection of Naegleria fowleri in surface water and sediment.

PubMed

Streby, Ashleigh; Mull, Bonnie J; Levy, Karen; Hill, Vincent R

2015-05-01

Naegleria fowleri is a thermophilic free-living ameba found in freshwater environments worldwide. It is the cause of a rare but potentially fatal disease in humans known as primary amebic meningoencephalitis. Established N. fowleri detection methods rely on conventional culture techniques and morphological examination followed by molecular testing. Multiple alternative real-time PCR assays have been published for rapid detection of Naegleria spp. and N. fowleri. Foursuch assays were evaluated for the detection of N. fowleri from surface water and sediment. The assays were compared for thermodynamic stability, analytical sensitivity and specificity, detection limits, humic acid inhibition effects, and performance with seeded environmental matrices. Twenty-one ameba isolates were included in the DNA panel used for analytical sensitivity and specificity analyses. N. fowleri genotypes I and III were used for method performance testing. Two of the real-time PCR assays were determined to yield similar performance data for specificity and sensitivity for detecting N. fowleri in environmental matrices.

Comparison of real-time PCR methods for the detection of Naegleria fowleri in surface water and sediment

PubMed Central

Streby, Ashleigh; Mull, Bonnie J.; Levy, Karen

2015-01-01

Naegleria fowleri is a thermophilic free-living ameba found in freshwater environments worldwide. It is the cause of a rare but potentially fatal disease in humans known as primary amebic meningoencephalitis. Established N. fowleri detection methods rely on conventional culture techniques and morphological examination followed by molecular testing. Multiple alternative real-time PCR assays have been published for rapid detection of Naegleria spp. and N. fowleri. Four such assays were evaluated for the detection of N. fowleri from surface water and sediment. The assays were compared for thermodynamic stability, analytical sensitivity and specificity, detection limits, humic acid inhibition effects, and performance with seeded environmental matrices. Twenty-one ameba isolates were included in the DNA panel used for analytical sensitivity and specificity analyses. N. fowleri genotypes I and III were used for method performance testing. Two of the real-time PCR assays were determined to yield similar performance data for specificity and sensitivity for detecting N. fowleri in environmental matrices. PMID:25855343

Parahydrogen-enhanced zero-field nuclear magnetic resonance

NASA Astrophysics Data System (ADS)

Theis, T.; Ganssle, P.; Kervern, G.; Knappe, S.; Kitching, J.; Ledbetter, M. P.; Budker, D.; Pines, A.

2011-07-01

Nuclear magnetic resonance, conventionally detected in magnetic fields of several tesla, is a powerful analytical tool for the determination of molecular identity, structure and function. With the advent of prepolarization methods and detection schemes using atomic magnetometers or superconducting quantum interference devices, interest in NMR in fields comparable to the Earth's magnetic field and below (down to zero field) has been revived. Despite the use of superconducting quantum interference devices or atomic magnetometers, low-field NMR typically suffers from low sensitivity compared with conventional high-field NMR. Here we demonstrate direct detection of zero-field NMR signals generated through parahydrogen-induced polarization, enabling high-resolution NMR without the use of any magnets. The sensitivity is sufficient to observe spectra exhibiting 13C-1H scalar nuclear spin-spin couplings (known as J couplings) in compounds with 13C in natural abundance, without the need for signal averaging. The resulting spectra show distinct features that aid chemical fingerprinting.

A Simple and Direct LC-MS Method for Determination of Genotoxic Impurity Hydroxylamine in Pharmaceutical compounds.

PubMed

Kumar, Thangarathinam; Ramya, Mohandass; Srinivasan, Viswanathan; Xavier, N

2017-08-01

Hydroxylamine is a known genotoxic impurity compound that needs to be controlled down to ppm level in pharmaceutical processes. It is difficult to detect using conventional analytical techniques due to its physio-chemical properties like lack of chromophore, low molecular weight, absence of carbon atom and high polarity. In addition to that, analysis of the pharmaceutical samples encounters considerable obstruction from matrix components that greatly overshadow the response of hydroxylamine. This study describes a simple, sensitive and direct Liquid Chromatographic-Mass Spectrometric method (LC-MS) for detection of hydroxylamine in pharmaceutical compounds. The LC-MS method was detected up to 0.008 ppm of hydroxylamine with S/N > 3.0 and quantified up to 0.025 ppm of hydroxylamine with S/N ratio >10.0. This validated method can be applied as a generic method to detect the hydroxylamine for pharmaceutical process control and drug substance release. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Analysis of Endocrine Disrupting Pesticides by Capillary GC with Mass Spectrometric Detection

PubMed Central

Matisová, Eva; Hrouzková, Svetlana

2012-01-01

Endocrine disrupting chemicals, among them many pesticides, alter the normal functioning of the endocrine system of both wildlife and humans at very low concentration levels. Therefore, the importance of method development for their analysis in food and the environment is increasing. This also covers contributions in the field of ultra-trace analysis of multicomponent mixtures of organic pollutants in complex matrices. With this fact conventional capillary gas chromatography (CGC) and fast CGC with mass spectrometric detection (MS) has acquired a real importance in the analysis of endocrine disrupting pesticide (EDP) residues. This paper provides an overview of GC methods, including sample preparation steps, for analysis of EDPs in a variety of matrices at ultra-trace concentration levels. Emphasis is put on separation method, mode of MS detection and ionization and obtained limits of detection and quantification. Analysis time is one of the most important aspects that should be considered in the choice of analytical methods for routine analysis. Therefore, the benefits of developed fast GC methods are important. PMID:23202677

The development of methods for the detection of Salmonella in chickens by a combination of immunomagnetic separation and PCRs.

PubMed

Dai, Fengying; Zhang, Miao; Xu, Dixin; Yang, Yin; Wang, Jiaxiao; Li, Mingzhen; Du, Meihong

2017-11-01

Micro- and nanoimmunomagnetic beads (MIMBs and NIMBs) used for immunomagnetic separation (IMS) with PCR were studied for the rapid detection of Salmonella. The capture efficiency of the two different IMBs was evaluated by a conventional plate counting method, and the binding pattern was studied using scanning electron microscopy. The specificity of the IMBs was tested with Salmonella, Shigella flexneri, enterohemorrhagic Escherichia coli O157:H7, and Listeria monocytogenes. By comparing the pre-enrichment IMS and the IMS enrichment steps with a 5.5-H enrichment time, this study developed a rapid and sensitive method for the detection of Salmonella in chicken. The method was implemented by IMS enrichment and PCR with MIMBs and NIMBs, with a total analysis time of 8 H. We showed that the method was sensitive based on NIMBs with a detection limit of 10° CFU for Salmonella in 25 g of chicken. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

Improvement of ion chromatography with ultraviolet photometric detection and comparison with conductivity detection for the determination of serum cations.

PubMed

Shintani, H

1985-05-31

Studies were made of the analytical conditions required for indirect photometric ion chromatography using ultraviolet photometric detection (UV method) for the determination of serum cations following a previously developed serum pre-treatment. The sensitivities of the conductivity detection (CD) and UV methods and the amounts of serum cations determined by both methods were compared. Attempts to improve the sensitivity of the conventional UV method are reported. It was found that the mobile phase previously reported by Small and Miller showed no quantitative response when more than 4 mM copper(II) sulphate pentahydrate was used. As a result, there was no significant difference in the amounts of serum cations shown by the CD and UV methods. However, by adding 0.5-5 mM cobalt(II) sulphate heptahydrate, nickel(II) sulphate hexahydrate, zinc(II) sulphate heptahydrate or cobalt(II) diammonium sulphate hexahydrate to 0.5-1.5 mM copper(II) sulphate pentahydrate, higher sensitivity and a quantitative response were attained.

Method for automatic localization of MR-visible markers using morphological image processing and conventional pulse sequences: feasibility for image-guided procedures.

PubMed

Busse, Harald; Trampel, Robert; Gründer, Wilfried; Moche, Michael; Kahn, Thomas

2007-10-01

To evaluate the feasibility and accuracy of an automated method to determine the 3D position of MR-visible markers. Inductively coupled RF coils were imaged in a whole-body 1.5T scanner using the body coil and two conventional gradient echo sequences (FLASH and TrueFISP) and large imaging volumes up to (300 mm(3)). To minimize background signals, a flip angle of approximately 1 degrees was used. Morphological 2D image processing in orthogonal scan planes was used to determine the 3D positions of a configuration of three fiducial markers (FMC). The accuracies of the marker positions and of the orientation of the plane defined by the FMC were evaluated at various distances r(M) from the isocenter. Fiducial marker detection with conventional equipment (pulse sequences, imaging coils) was very reliable and highly reproducible over a wide range of experimental conditions. For r(M)

Epidermal segmentation in high-definition optical coherence tomography.

PubMed

Li, Annan; Cheng, Jun; Yow, Ai Ping; Wall, Carolin; Wong, Damon Wing Kee; Tey, Hong Liang; Liu, Jiang

2015-01-01

Epidermis segmentation is a crucial step in many dermatological applications. Recently, high-definition optical coherence tomography (HD-OCT) has been developed and applied to imaging subsurface skin tissues. In this paper, a novel epidermis segmentation method using HD-OCT is proposed in which the epidermis is segmented by 3 steps: the weighted least square-based pre-processing, the graph-based skin surface detection and the local integral projection-based dermal-epidermal junction detection respectively. Using a dataset of five 3D volumes, we found that this method correlates well with the conventional method of manually marking out the epidermis. This method can therefore serve to effectively and rapidly delineate the epidermis for study and clinical management of skin diseases.

The Safety Course Design and Operations of Composite Overwrapped Pressure Vessels (COPV)

NASA Technical Reports Server (NTRS)

Saulsberry, Regor; Prosser, William

2015-01-01

Following a Commercial Launch Vehicle On-Pad COPV (Composite Overwrapped Pressure Vessels) failure, a request was received by the NESC (NASA Engineering and Safety Center) June 14, 2014. An assessment was approved July 10, 2014, to develop and assess the capability of scanning eddy current (EC) nondestructive evaluation (NDE) methods for mapping thickness and inspection for flaws. Current methods could not identify thickness reduction from necking and critical flaw detection was not possible with conventional dye penetrant (PT) methods, so sensitive EC scanning techniques were needed. Developmental methods existed, but had not been fully developed, nor had the requisite capability assessment (i.e., a POD (Probability of Detection) study) been performed.

Microtiter format for simultaneous multianalyte detection and development of a PCR-chemiluminescent enzyme immunoassay for typing human papillomavirus DNAs.

PubMed

Roda, Aldo; Mirasoli, Mara; Venturoli, Simona; Cricca, Monica; Bonvicini, Francesca; Baraldini, Mario; Pasini, Patrizia; Zerbini, Marialuisa; Musiani, Monica

2002-10-01

To allow multianalyte binding assays, we have developed a novel polystyrene microtiter plate containing 24 main wells, each divided into 7 subwells. We explored its clinical potential by developing a PCR-chemiluminescent immunoassay (PCR-CLEIA) for simultaneous detection and typing of seven high oncogenic risk human papillomavirus (HPV) DNAs in one well. Seven different oligonucleotide probes, each specific for a high-risk HPV genotype, were separately immobilized in the subwells. Subsequently, a digoxigenin-labeled consensus PCR amplification product was added to the main well. The PCR product hybridized to the immobilized probe corresponding to its genotype and was subsequently detected by use of a peroxidase-labeled anti-digoxigenin antibody and chemiluminescence imaging with an ultrasensitive charge-coupled device camera. Results obtained for 50 cytologic samples were compared with those obtained with a conventional colorimetric PCR-ELISA. The method was specific and allowed detection of 50 genome copies of HPV 16, 18, 33, and 58, and 100 genome copies of HPV 31, 35, and 45. Intra- and interassay CVs for the method were 5.6% and 7.9%, respectively. All results obtained for clinical samples were confirmed by the conventional PCR-ELISA. PCR-CLEIA allows rapid, single-tube simultaneous detection and typing of seven high-risk HPV DNAs with small reagent volumes. The principle appears applicable to the development of other single-tube panels of tests.

[A Duplex PCR Method for Detection of Babesia caballi and Theileria equi].

PubMed

Zhang, Yang; Zhang, Yu-ting; Wang, Zhen-bao; Bolati; Li, Hai; Bayinchahan

2015-04-01

To develop a duplex PCR assay for detection of Babesia caballi and Theileria equi. Two pairs of primers were designed according to the BC48 gene of B. caballi and 18 s rRNA gene of T. equi, and a duplex PCR assay was developed by the optimization of reaction conditions. The specificity, sensitivity and reliability of the method were tested. The horse blood samples of suspected cases were collected from Yili region, and detected by the duplex PCR, microspopy, conventional PCR, and fluorescence quantitative PCR, and the results were compared. Using the duplex PCR assay, the specific fragments of 155 bp and 280 bp were amplified from DNA samples of B. caballi and T. equi, respectively. No specific fragment was amplified from DNA samples of B. bigemina, Theilerdia annulata, Theilerdia sergenti, Toxoplasma gondii, Neospora caninum, and Trypanosoma evansi. The limit of detection was 4.85 x 10(5) copies/L for B. caballi DNA and 4.85 x 10(4) copies/µl for T. equi DNA, respectively. Among the 24 blood samples, 11 were found B. caballi-positive by the duplex PCR assay, and 18 were T. equi-positive. The coincidence rate of microscopy, conventional PCR, and fluorescence quantitative PCR with duplex PCR was 91.7% (22/24), 95.8% (23/24), and 95.8% (23/24), respectively. A duplex PCR assay for simultaneous detection of B. caballi and T. equi is established.

Terahertz Imaging and Backscatter Radiography Probability of Detection Study for Space Shuttle Foam Inspections

NASA Technical Reports Server (NTRS)

Ussery, Warren; Johnson, Kenneth; Walker, James; Rummel, Ward

2008-01-01

This slide presentation reviews the use of terahertz imaging and Backscatter Radiography in a probability of detection study of the foam on the external tank (ET) shedding and damaging the shuttle orbiter. Non-destructive Examination (NDE) is performed as one method of preventing critical foam debris during the launch. Conventional NDE methods for inspection of the foam are assessed and the deficiencies are reviewed. Two methods for NDE inspection are reviewed: Backscatter Radiography (BSX) and Terahertz (THZ) Imaging. The purpose of the Probability of Detection (POD) study was to assess performance and reliability of the use of BSX and or THZ as an appropriate NDE method. The study used a test article with inserted defects, and a sample of blanks included to test for false positives. The results of the POD study are reported.

Fault detection and diagnosis in asymmetric multilevel inverter using artificial neural network

NASA Astrophysics Data System (ADS)

Raj, Nithin; Jagadanand, G.; George, Saly

2018-04-01

The increased component requirement to realise multilevel inverter (MLI) fallout in a higher fault prospect due to power semiconductors. In this scenario, efficient fault detection and diagnosis (FDD) strategies to detect and locate the power semiconductor faults have to be incorporated in addition to the conventional protection systems. Even though a number of FDD methods have been introduced in the symmetrical cascaded H-bridge (CHB) MLIs, very few methods address the FDD in asymmetric CHB-MLIs. In this paper, the gate-open circuit FDD strategy in asymmetric CHB-MLI is presented. Here, a single artificial neural network (ANN) is used to detect and diagnose the fault in both binary and trinary configurations of the asymmetric CHB-MLIs. In this method, features of the output voltage of the MLIs are used as to train the ANN for FDD method. The results prove the validity of the proposed method in detecting and locating the fault in both asymmetric MLI configurations. Finally, the ANN response to the input parameter variation is also analysed to access the performance of the proposed ANN-based FDD strategy.

Laser-Based Pedestrian Tracking in Outdoor Environments by Multiple Mobile Robots

PubMed Central

Ozaki, Masataka; Kakimuma, Kei; Hashimoto, Masafumi; Takahashi, Kazuhiko

2012-01-01

This paper presents an outdoors laser-based pedestrian tracking system using a group of mobile robots located near each other. Each robot detects pedestrians from its own laser scan image using an occupancy-grid-based method, and the robot tracks the detected pedestrians via Kalman filtering and global-nearest-neighbor (GNN)-based data association. The tracking data is broadcast to multiple robots through intercommunication and is combined using the covariance intersection (CI) method. For pedestrian tracking, each robot identifies its own posture using real-time-kinematic GPS (RTK-GPS) and laser scan matching. Using our cooperative tracking method, all the robots share the tracking data with each other; hence, individual robots can always recognize pedestrians that are invisible to any other robot. The simulation and experimental results show that cooperating tracking provides the tracking performance better than conventional individual tracking does. Our tracking system functions in a decentralized manner without any central server, and therefore, this provides a degree of scalability and robustness that cannot be achieved by conventional centralized architectures. PMID:23202171

Real-time RT-PCR, a necessary tool to support the diagnosis and surveillance of rotavirus in Mexico.

PubMed

De La Cruz Hernández, Sergio Isaac; Anaya Molina, Yazmin; Gómez Santiago, Fabián; Terán Vega, Heidi Lizbeth; Monroy Leyva, Elda; Méndez Pérez, Héctor; García Lozano, Herlinda

2018-04-01

Rotavirus produces diarrhea in children under 5 years old. Most of those conventional methods such as polyacrylamide gel electrophoresis (PAGE) and reverse transcription-polymerase chain reaction (RT-PCR) have been used for rotavirus detection. However, these techniques need a multi-step process to get the results. In comparison with conventional methods, the real-time RT-PCR is a highly sensitive method, which allows getting the results in only one day. In this study a real-time RT-PCR assay was tested using a panel of 440 samples from patients with acute gastroenteritis, and characterized by PAGE and RT-PCR. The results show that the real-time RT-PCR detected rotavirus from 73% of rotavirus-negative samples analyzed by PAGE and RT-PCR; thus, the percentage of rotavirus-positive samples increased to 81%. The results indicate that this real-time RT-PCR should be part of a routine analysis, and as a support of the diagnosis of rotavirus in Mexico. Copyright © 2017 Elsevier Inc. All rights reserved.

A longitudinal study of Campylobacter distribution in a turkey production chain

PubMed Central

Perko-Mäkelä, Päivikki; Isohanni, Pauliina; Katzav, Marianne; Lund, Marianne; Hänninen, Marja-Liisa; Lyhs, Ulrike

2009-01-01

Background Campylobacter is the most common cause of bacterial enteritis worldwide. Handling and eating of contaminated poultry meat has considered as one of the risk factors for human campylobacteriosis.Campylobacter contamination can occur at all stages of a poultry production cycle. The objective of this study was to determine the occurrence of Campylobacter during a complete turkey production cycle which lasts for 1,5 years of time. For detection of Campylobacter, a conventional culture method was compared with a PCR method. Campylobacter isolates from different types of samples have been identified to the species level by a multiplex PCR assay. Methods Samples (N = 456) were regularly collected from one turkey parent flock, the hatchery, six different commercial turkey farms and from 11 different stages at the slaughterhouse. For the detection of Campylobacter, a conventional culture and a PCR method were used. Campylobacter isolates (n = 143) were identified to species level by a multiplex PCR assay. Results No Campylobacter were detected in either the samples from the turkey parent flock or from hatchery samples using the culture method. PCR detected Campylobacter DNA in five faecal samples and one fluff and eggshell sample. Six flocks out of 12 commercial turkey flocks where found negative at the farm level but only two were negative at the slaughterhouse. Conclusion During the brooding period Campylobacter might have contact with the birds without spreading of the contamination within the flock. Contamination of working surfaces and equipment during slaughter of a Campylobacter positive turkey flock can persist and lead to possible contamination of negative flocks even after the end of the day's cleaning and desinfection. Reduction of contamination at farm by a high level of biosecurity control and hygiene may be one of the most efficient ways to reduce the amount of contaminated poultry meat in Finland. Due to the low numbers of Campylobacter in the Finnish turkey production chain, enrichment PCR seems to be the optimal detection method here. PMID:19348687

Relative spectral response calibration using Ti plasma lines

NASA Astrophysics Data System (ADS)

Teng, FEI; Congyuan, PAN; Qiang, ZENG; Qiuping, WANG; Xuewei, DU

2018-04-01

This work introduces the branching ratio (BR) method for determining relative spectral responses, which are needed routinely in laser induced breakdown spectroscopy (LIBS). Neutral and singly ionized Ti lines in the 250–498 nm spectral range are investigated by measuring laser-induced micro plasma near a Ti plate and used to calculate the relative spectral response of an entire LIBS detection system. The results are compared with those of the conventional relative spectral response calibration method using a tungsten halogen lamp, and certain lines available for the BR method are selected. The study supports the common manner of using BRs to calibrate the detection system in LIBS setups.

Measurement of pattern roughness and local size variation using CD-SEM: current status

NASA Astrophysics Data System (ADS)

Fukuda, Hiroshi; Kawasaki, Takahiro; Kawada, Hiroki; Sakai, Kei; Kato, Takashi; Yamaguchi, Satoru; Ikota, Masami; Momonoi, Yoshinori

2018-03-01

Measurement of line edge roughness (LER) is discussed from four aspects: edge detection, PSD prediction, sampling strategy, and noise mitigation, and general guidelines and practical solutions for LER measurement today are introduced. Advanced edge detection algorithms such as wave-matching method are shown effective for robustly detecting edges from low SNR images, while conventional algorithm with weak filtering is still effective in suppressing SEM noise and aliasing. Advanced PSD prediction method such as multi-taper method is effective in suppressing sampling noise within a line edge to analyze, while number of lines is still required for suppressing line to line variation. Two types of SEM noise mitigation methods, "apparent noise floor" subtraction method and LER-noise decomposition using regression analysis are verified to successfully mitigate SEM noise from PSD curves. These results are extended to LCDU measurement to clarify the impact of SEM noise and sampling noise on LCDU.

Contrast-enhanced optical coherence tomography with picomolar sensitivity for functional in vivo imaging

NASA Astrophysics Data System (ADS)

Liba, Orly; Sorelle, Elliott D.; Sen, Debasish; de La Zerda, Adam

2016-03-01

Optical Coherence Tomography (OCT) enables real-time imaging of living tissues at cell-scale resolution over millimeters in three dimensions. Despite these advantages, functional biological studies with OCT have been limited by a lack of exogenous contrast agents that can be distinguished from tissue. Here we report an approach to functional OCT imaging that implements custom algorithms to spectrally identify unique contrast agents: large gold nanorods (LGNRs). LGNRs exhibit 110-fold greater spectral signal per particle than conventional GNRs, which enables detection of individual LGNRs in water and concentrations as low as 250 pM in the circulation of living mice. This translates to ~40 particles per imaging voxel in vivo. Unlike previous implementations of OCT spectral detection, the methods described herein adaptively compensate for depth and processing artifacts on a per sample basis. Collectively, these methods enable high-quality noninvasive contrast-enhanced imaging of OCT in living subjects, including detection of tumor microvasculature at twice the depth achievable with conventional OCT. Additionally, multiplexed detection of spectrally-distinct LGNRs was demonstrated to observe discrete patterns of lymphatic drainage and identify individual lymphangions and lymphatic valve functional states. These capabilities provide a powerful platform for molecular imaging and characterization of tissue noninvasively at cellular resolution, called MOZART.

Tuning time-frequency methods for the detection of metered HF speech

NASA Astrophysics Data System (ADS)

Nelson, Douglas J.; Smith, Lawrence H.

2002-12-01

Speech is metered if the stresses occur at a nearly regular rate. Metered speech is common in poetry, and it can occur naturally in speech, if the speaker is spelling a word or reciting words or numbers from a list. In radio communications, the CQ request, call sign and other codes are frequently metered. In tactical communications and air traffic control, location, heading and identification codes may be metered. Moreover metering may be expected to survive even in HF communications, which are corrupted by noise, interference and mistuning. For this environment, speech recognition and conventional machine-based methods are not effective. We describe Time-Frequency methods which have been adapted successfully to the problem of mitigation of HF signal conditions and detection of metered speech. These methods are based on modeled time and frequency correlation properties of nearly harmonic functions. We derive these properties and demonstrate a performance gain over conventional correlation and spectral methods. Finally, in addressing the problem of HF single sideband (SSB) communications, the problems of carrier mistuning, interfering signals, such as manual Morse, and fast automatic gain control (AGC) must be addressed. We demonstrate simple methods which may be used to blindly mitigate mistuning and narrowband interference, and effectively invert the fast automatic gain function.

Endoscopic nasopharyngeal exploration at the end of conventional curettage adenoidectomy.

PubMed

Abdel-Aziz, Mosaad

2012-03-01

Adenoid hypertrophy (AH) is a common cause of airway obstruction in children and its recurrence after conventional curettage adenoidectomy is not rare. The purpose of this study is to assess the efficacy of endoscopic nasopharyngeal exploration at the end of curettage adenoidectomy on decreasing the incidence of adenoid re-hypertrophy. Three hundred and fifty children diagnosed as having AH, underwent conventional curettage adenoidectomy by a single surgeon. The cases were randomly divided into two equal groups A and B, group B were further subjected to nasopharyngeal exploration by the nasal endoscope after removal of their adenoids with cauterization of any visible residuals, while group A were not subjected to this endoscopic maneuver. Follow-up was carried out for at least 2 years; flexible nasopharyngoscopy was used for detection of recurrent AH. Cases that were not subjected to endoscopic nasopharyngeal exploration (group A) showed a high recurrence rate (6.6%), while explored cases (group B) showed a low incidence of recurrence (1.18%). Most recurrence of group A (6%) was detected within the first year of the follow-up period which may indicate re-growth of residual adenoidal tissues that were missed during conventional curettage adenoidectomy. Endoscopic nasopharyngeal exploration at the end of conventional curettage adenoidectomy is a useful method in decreasing the incidence of recurrent AH.

Innovations in diagnostic imaging of localized prostate cancer.

PubMed

Pummer, Karl; Rieken, Malte; Augustin, Herbert; Gutschi, Thomas; Shariat, Shahrokh F

2014-08-01

In recent years, various imaging modalities have been developed to improve diagnosis, staging, and localization of early-stage prostate cancer (PCa). A MEDLINE literature search of the time frame between 01/2007 and 06/2013 was performed on imaging of localized PCa. Conventional transrectal ultrasound (TRUS) is mainly used to guide prostate biopsy. Contrast-enhanced ultrasound is based on the assumption that PCa tissue is hypervascularized and might be better identified after intravenous injection of a microbubble contrast agent. However, results on its additional value for cancer detection are controversial. Computer-based analysis of the transrectal ultrasound signal (C-TRUS) appears to detect cancer in a high rate of patients with previous biopsies. Real-time elastography seems to have higher sensitivity, specificity, and positive predictive value than conventional TRUS. However, the method still awaits prospective validation. The same is true for prostate histoscanning, an ultrasound-based method for tissue characterization. Currently, multiparametric MRI provides improved tissue visualization of the prostate, which may be helpful in the diagnosis and targeting of prostate lesions. However, most published series are small and suffer from variations in indication, methodology, quality, interpretation, and reporting. Among ultrasound-based techniques, real-time elastography and C-TRUS seem the most promising techniques. Multiparametric MRI appears to have advantages over conventional T2-weighted MRI in the detection of PCa. Despite these promising results, currently, no recommendation for the routine use of these novel imaging techniques can be made. Prospective studies defining the value of various imaging modalities are urgently needed.

A comparison of PCR assays for beak and feather disease virus and high resolution melt (HRM) curve analysis of replicase associated protein and capsid genes.

PubMed

Das, Shubhagata; Sarker, Subir; Ghorashi, Seyed Ali; Forwood, Jade K; Raidal, Shane R

2016-11-01

Beak and feather disease virus (BFDV) threatens a wide range of endangered psittacine birds worldwide. In this study, we assessed a novel PCR assay and genetic screening method using high-resolution melt (HRM) curve analysis for BFDV targeting the capsid (Cap) gene (HRM-Cap) alongside conventional PCR detection as well as a PCR method that targets a much smaller fragment of the virus genome in the replicase initiator protein (Rep) gene (HRM-Rep). Limits of detection, sensitivity, specificity and discriminatory power for differentiating BFDV sequences were compared. HRM-Cap had a high positive predictive value and could readily differentiate between a reference genotype and 17 other diverse BFDV genomes with more discriminatory power (genotype confidence percentage) than HRM-Rep. Melt curve profiles generated by HRM-Cap correlated with unique DNA sequence profiles for each individual test genome. The limit of detection of HRM-Cap was lower (2×10 -5 ng/reaction or 48 viral copies) than that for both HRM-Rep and conventional BFDV PCR which had similar sensitivity (2×10 -6 ng or 13 viral copies/reaction). However, when used in a diagnostic setting with 348 clinical samples there was strong agreement between HRM-Cap and conventional PCR (kappa=0.87, P<0.01, 98% specificity) and HRM-Cap demonstrated higher specificity (99.9%) than HRM-Rep (80.3%). Copyright © 2016 Elsevier B.V. All rights reserved.

Novel laser Doppler flowmeter for pulpal blood flow measurements

NASA Astrophysics Data System (ADS)

Zang, De Yu; Millerd, James E.; Wilder-Smith, Petra B. B.; Arrastia-Jitosho, Anna-Marie A.

1996-04-01

We have proposed and experimentally demonstrated a new configuration of laser Doppler flowmetry for dental pulpal blood flow measurements. To date, the vitality of a tooth can be determined only by subjective thermal or electric tests, which are of questionable reliability and may induced pain in patient. Non-invasive techniques for determining pulpal vascular reactions to injury, treatment, and medication are in great demand. The laser Doppler flowmetry technique is non-invasive; however, clinical studies have shown that when used to measure pulpal blood flow the conventional back-scattering Doppler method suffers from low signal-to-noise ratio (SNR) and unreliable flux readings rendering it impossible to calibrate. A simplified theoretical model indicates that by using a forward scattered geometry the detected signal has a much higher SNR and can be calibrated. The forward scattered signal is readily detectable due to the fact that teeth are relatively thin organs with moderate optical loss. A preliminary experiment comparing forward scattered detection with conventional back- scattered detection was carried out using an extracted human molar. The results validated the findings of the simple theoretical model and clearly showed the utility of the forward scattering geometry. The back-scattering method had readings that fluctuated by as much as 187% in response to small changes in sensor position relative to the tooth. The forward scattered method had consistent readings (within 10%) that were independent of the sensor position, a signal-to-noise ratio that was at least 5.6 times higher than the back-scattering method, and a linear response to flow rate.

Electrochemical detection of influenza virus H9N2 based on both immunomagnetic extraction and gold catalysis using an immobilization-free screen printed carbon microelectrode.

PubMed

Sayhi, Maher; Ouerghi, Oussama; Belgacem, Kamel; Arbi, Marwa; Tepeli, Yudum; Ghram, Abdeljalil; Anik, Ülkü; Österlund, Lars; Laouini, Dhafer; Diouani, Mohamed Fethi

2018-06-01

Influenza is a viral infectious disease considered as a source of many health problems and enormous socioeconomic disruptions. Conventional methods are inadequate for in-field detection of the virus and generally suffer from being laborious and time-consuming. Thus, studies aiming to develop effective alternatives to conventional methods are urgently needed. In this work, we developed an approach for the isolation and detection of influenza A virus subtype H9N2. For this aim, two specific influenza receptors were used. The first, anti-matrix protein 2 (M2) antibody, was attached to iron magnetic nanoparticles (MNPs) and used for the isolation of the virus from allantoic fluid. The second biomolecule, Fetuin A, was attached to an electrochemical detectable label, gold nanoparticles (AuNPs), and used to detect the virus tacking advantage from fetuin-hemagglutinin interaction. The MNP-Influenza virus-AuNP formed complex was isolated and treated by an acid solution then the collected gold nanoparticles were deposited onto a screen printed carbon electrode. AuNPs catalyzes the hydrogen ions reduction in acidic medium while applying an appropriate potential, and the generated current signal was proportional to the virus titer. This approach allows the rapid detection of influenza virus A/H9N2 at a less than 16 HAU titer. Copyright © 2018 Elsevier B.V. All rights reserved.

A microfluidic approach for hemoglobin detection in whole blood

NASA Astrophysics Data System (ADS)

Taparia, Nikita; Platten, Kimsey C.; Anderson, Kristin B.; Sniadecki, Nathan J.

2017-10-01

Diagnosis of anemia relies on the detection of hemoglobin levels in a blood sample. Conventional blood analyzers are not readily available in most low-resource regions where anemia is prevalent, so detection methods that are low-cost and point-of-care are needed. Here, we present a microfluidic approach to measure hemoglobin concentration in a sample of whole blood. Unlike conventional approaches, our microfluidic approach does not require hemolysis. We detect the level of hemoglobin in a blood sample optically by illuminating the blood in a microfluidic channel at a peak wavelength of 540 nm and measuring its absorbance using a CMOS sensor coupled with a lens to magnify the image onto the detector. We compare measurements in microchannels with channel heights of 50 and 115 μm and found the channel with the 50 μm height provided a better range of detection. Since we use whole blood and not lysed blood, we fit our data to an absorption model that includes optical scattering in order to obtain a calibration curve for our system. Based on this calibration curve and data collected, we can measure hemoglobin concentration within 1 g/dL for severe cases of anemia. In addition, we measured optical density for blood flowing at a shear rate of 500 s-1 and observed it did not affect the nonlinear model. With this method, we provide an approach that uses microfluidic detection of hemoglobin levels that can be integrated with other microfluidic approaches for blood analysis.

Next-generation sequencing-based method shows increased mutation detection sensitivity in an Indian retinoblastoma cohort

PubMed Central

Singh, Jaya; Mishra, Avshesh; Pandian, Arunachalam Jayamuruga; Mallipatna, Ashwin C.; Khetan, Vikas; Sripriya, S.; Kapoor, Suman; Agarwal, Smita; Sankaran, Satish; Katragadda, Shanmukh; Veeramachaneni, Vamsi; Hariharan, Ramesh; Subramanian, Kalyanasundaram

2016-01-01

Purpose Retinoblastoma (Rb) is the most common primary intraocular cancer of childhood and one of the major causes of blindness in children. India has the highest number of patients with Rb in the world. Mutations in the RB1 gene are the primary cause of Rb, and heterogeneous mutations are distributed throughout the entire length of the gene. Therefore, genetic testing requires screening of the entire gene, which by conventional sequencing is time consuming and expensive. Methods In this study, we screened the RB1 gene in the DNA isolated from blood or saliva samples of 50 unrelated patients with Rb using the TruSight Cancer panel. Next-generation sequencing (NGS) was done on the Illumina MiSeq platform. Genetic variations were identified using the Strand NGS software and interpreted using the StrandOmics platform. Results We were able to detect germline pathogenic mutations in 66% (33/50) of the cases, 12 of which were novel. We were able to detect all types of mutations, including missense, nonsense, splice site, indel, and structural variants. When we considered bilateral Rb cases only, the mutation detection rate increased to 100% (22/22). In unilateral Rb cases, the mutation detection rate was 30% (6/20). Conclusions Our study suggests that NGS-based approaches increase the sensitivity of mutation detection in the RB1 gene, making it fast and cost-effective compared to the conventional tests performed in a reflex-testing mode. PMID:27582626

Identification and quantification of three genetically modified insect resistant cotton lines using conventional and TaqMan real-time polymerase chain reaction methods.

PubMed

Yang, Litao; Pan, Aihu; Zhang, Kewei; Guo, Jinchao; Yin, Changsong; Chen, Jianxiu; Huang, Cheng; Zhang, Dabing

2005-08-10

As the genetically modified organisms (GMOs) labeling policies are issued in many countries, qualitative and quantitative polymerase chain reaction (PCR) techniques are increasingly used for the detection of genetically modified (GM) crops in foods. Qualitative PCR and TaqMan real-time quantitative PCR methods to detect and identify three varieties of insect resistant cotton, i.e., Mon531 cotton (Monsanto Co.) and GK19 and SGK321 cottons (Chinese Academy of Agricultural Sciences), which were approved for commercialization in China, were developed in this paper. Primer pairs specific to inserted DNAs, such as Cowpea trypsin inhibitor (CpTI) gene of SGK321 cotton and the specific junction DNA sequences containing partial Cry1A(c) gene and NOS terminator of Mon531, GK19, and SGK321 cotton varieties were designed to conduct the identified PCR assays. In conventional specific identified PCR assays, the limit of detection (LOD) was 0.05% for Mon531, GK19, or SGK321 in 100 ng of cotton genomic DNA for one reaction. Also, the multiplex PCR method for screening the three GM cottons was also established, which could save time and cost in practical detection. Furthermore, a real-time quantitative PCR assay based on TaqMan chemistry for detection of insect resistant gene, Cry1A(c), was developed. This assay also featured the use of a standard plasmid as a reference molecule, which contained both a specific region of the transgene Cry1A(c) and an endogenous stearoyl-acyl carrier protein desaturase (Sad1) gene of the cotton. In quantitative PCR assay, the quantification range was from 0.01 to 100% in 100 ng of the genome DNA template, and in the detection of 1.0, 3.0, and 5.0% levels of three insect resistant cotton lines, respectively, all of the relative standard deviations (RSDs) were less than 8.2% except for the GM cotton samples with 1.0% Mon531 or GK19, which meant that our real-time PCR assays involving the use of reference molecule were reliable and practical for GM insect resistant cottons quantification. All of these results indicated that our established conventional and TaqMan real-time PCR assays were applicable to detect the three insect resistant cottons qualitatively and quantitatively.

Design of Passive Power Filter for Hybrid Series Active Power Filter using Estimation, Detection and Classification Method

NASA Astrophysics Data System (ADS)

Swain, Sushree Diptimayee; Ray, Pravat Kumar; Mohanty, K. B.

2016-06-01

This research paper discover the design of a shunt Passive Power Filter (PPF) in Hybrid Series Active Power Filter (HSAPF) that employs a novel analytic methodology which is superior than FFT analysis. This novel approach consists of the estimation, detection and classification of the signals. The proposed method is applied to estimate, detect and classify the power quality (PQ) disturbance such as harmonics. This proposed work deals with three methods: the harmonic detection through wavelet transform method, the harmonic estimation by Kalman Filter algorithm and harmonic classification by decision tree method. From different type of mother wavelets in wavelet transform method, the db8 is selected as suitable mother wavelet because of its potency on transient response and crouched oscillation at frequency domain. In harmonic compensation process, the detected harmonic is compensated through Hybrid Series Active Power Filter (HSAPF) based on Instantaneous Reactive Power Theory (IRPT). The efficacy of the proposed method is verified in MATLAB/SIMULINK domain and as well as with an experimental set up. The obtained results confirm the superiority of the proposed methodology than FFT analysis. This newly proposed PPF is used to make the conventional HSAPF more robust and stable.

Detection of alpha particles using DNA/Al Schottky junctions

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

Al-Ta'ii, Hassan Maktuff Jaber, E-mail: hassankirkukly@gmail.com, E-mail: vengadeshp@um.edu.my; Department of Physics, Faculty of Science, University of Al-Muthana, Al-Muthana 66001; Periasamy, Vengadesh, E-mail: hassankirkukly@gmail.com, E-mail: vengadeshp@um.edu.my

2015-09-21

Deoxyribonucleic acid or DNA can be utilized in an organic-metallic rectifying structure to detect radiation, especially alpha particles. This has become much more important in recent years due to crucial environmental detection needs in both peace and war. In this work, we fabricated an aluminum (Al)/DNA/Al structure and generated current–voltage characteristics upon exposure to alpha radiation. Two models were utilized to investigate these current profiles; the standard conventional thermionic emission model and Cheung and Cheung's method. Using these models, the barrier height, Richardson constant, ideality factor and series resistance of the metal-DNA-metal structure were analyzed in real time. The barriermore » height, Φ value calculated using the conventional method for non-radiated structure was 0.7149 eV, increasing to 0.7367 eV after 4 min of radiation. Barrier height values were observed to increase after 20, 30 and 40 min of radiation, except for 6, 8, and 10 min, which registered a decrease of about 0.67 eV. This was in comparison using Cheung and Cheung's method, which registered 0.6983 eV and 0.7528 eV for the non-radiated and 2 min of radiation, respectively. The barrier height values, meanwhile, were observed to decrease after 4 (0.61 eV) to 40 min (0.6945 eV). The study shows that conventional thermionic emission model could be practically utilized for estimating the diode parameters including the effect of series resistance. These changes in the electronic properties of the Al/DNA/Al junctions could therefore be utilized in the manufacture of sensitive alpha particle sensors.« less

Detection of Gastrointestinal Pathogens from Stool Samples on Hemoccult Cards by Multiplex PCR.

PubMed

Alberer, Martin; Schlenker, Nicklas; Bauer, Malkin; Helfrich, Kerstin; Mengele, Carolin; Löscher, Thomas; Nothdurft, Hans Dieter; Bretzel, Gisela; Beissner, Marcus

2017-01-01

Purpose . Up to 30% of international travelers are affected by travelers' diarrhea (TD). Reliable data on the etiology of TD is lacking. Sufficient laboratory capacity at travel destinations is often unavailable and transporting conventional stool samples to the home country is inconvenient. We evaluated the use of Hemoccult cards for stool sampling combined with a multiplex PCR for the detection of model viral, bacterial, and protozoal TD pathogens. Methods . Following the creation of serial dilutions for each model pathogen, last positive dilution steps (LPDs) and thereof calculated last positive sample concentrations (LPCs) were compared between conventional stool samples and card samples. Furthermore, card samples were tested after a prolonged time interval simulating storage during a travel duration of up to 6 weeks. Results . The LPDs/LPCs were comparable to testing of conventional stool samples. After storage on Hemoccult cards, the recovery rate was 97.6% for C. jejuni , 100% for E . histolytica , 97.6% for norovirus GI, and 100% for GII. Detection of expected pathogens was possible at weekly intervals up to 42 days. Conclusion . Stool samples on Hemoccult cards stored at room temperature can be used in combination with a multiplex PCR as a reliable tool for testing of TD pathogens.

Polyacrylonitrile Nanofiber-Based Quartz Crystal Microbalance for Sensitive Detection of Safrole

PubMed Central

Julian, Trisna; Hidayat, Shidiq Nur; Suyono, Eko Agus

2018-01-01

Safrole is the main precursor for producing the amphetamine-type stimulant (ATS) drug, N-methyl-3,4-methylenedioxyamphetamine (MDMA), also known as ecstasy. We devise a polyacrylonitrile (PAN) nanofiber-based quartz crystal microbalance (QCM) for detecting safrole. The PAN nanofibers were fabricated by direct electrospinning to modify the QCM chips. The PAN nanofiber on the QCM chips has a diameter of 240 ± 10 nm. The sensing of safrole by QCM modified with PAN nanofiber shows good reversibility and an apparent sensitivity of 4.6 Hz·L/mg. The proposed method is simple, inexpensive, and convenient for detecting safrole, and can be an alternative to conventional instrumental analytical methods for general volatile compounds. PMID:29642565

Quality characteristics of battered and fried chicken: comparison of pressure frying and conventional frying.

PubMed

Das, Rashmi; Pawar, Deepthi P; Modi, Vinod Kumar

2013-04-01

The marinated and battered chicken leg meat and breast meat were pressure fried and their physico-chemical qualities were compared to the conventional fried product (open pan deep fat frying). Shrinkage due to frying process was significantly lesser in case of pressure fried leg meat (PLM) and breast meat (PBM) as compared to products prepared by conventional frying leg meat (CLM) and breast meat (CBM). Also, juiciness of pressure fried chicken products was superior (p ≤ 0.05) than fried products obtained by the conventional method. PLM and PBM had lower fat content (p ≤ 0.05) compared to conventionally fried CLM and CBM. Lipid oxidation was higher (p ≤ 0.05) in conventional frying as compared to pressure frying. Irrespective of the type of chicken meat, conventionally fried meat required higher shear force as compared to pressure fried products. Salmonella, Staphylococcus aureus, Shigella and E. coli were not detected. The study indicates the usefulness and superiority of pressure frying in comparison to conventional deep fat frying.

Edge Detection Method Based on Neural Networks for COMS MI Images

NASA Astrophysics Data System (ADS)

Lee, Jin-Ho; Park, Eun-Bin; Woo, Sun-Hee

2016-12-01

Communication, Ocean And Meteorological Satellite (COMS) Meteorological Imager (MI) images are processed for radiometric and geometric correction from raw image data. When intermediate image data are matched and compared with reference landmark images in the geometrical correction process, various techniques for edge detection can be applied. It is essential to have a precise and correct edged image in this process, since its matching with the reference is directly related to the accuracy of the ground station output images. An edge detection method based on neural networks is applied for the ground processing of MI images for obtaining sharp edges in the correct positions. The simulation results are analyzed and characterized by comparing them with the results of conventional methods, such as Sobel and Canny filters.

A Novel QEPAS with Microresonator in the Open Environment

NASA Astrophysics Data System (ADS)

Lin, Cheng; Zhu, Yong; Wei, Wei; Wang, Ning; Bao, Weiyi

2013-09-01

An improved quartz-enhanced photoacoustic spectroscopy (QEPAS) sensing system for trace gas detection is proposed. The optical fiber Fabry-Perot (F-P) demodulation method is used to replace the conventional electrical one in the QEPAS system. The experimental QEPAS system, which has a microresonator consisting of two stainless steel tubes with a length of 2.3 mm and an inner diameter of 0.9 mm, is implemented to detect the absorption of water vapor in the open environment. The structure parameters of the quartz tuning fork (QTF) are optimized in order to make the sensing system work more stably and reliably. Demonstration experiments are carried out. The vibration signal of the QTF was picked up by the optical fiber F-P demodulator and the conventional electrical scheme at the same time. Normalized noise equivalent absorption coefficients of and are obtained, respectively. The experimental result demonstrates that the sensitivity of the improved QEPAS sensing system with an optical fiber F-P demodulator is about 5.9 times higher than that of the conventional QEPAS system.

On-line focusing of flavin derivatives using Dynamic pH junction-sweeping capillary electrophoresis with laser-induced fluorescence detection.

PubMed

Britz-McKibbin, Philip; Otsuka, Koji; Terabe, Shigeru

2002-08-01

Simple yet effective methods to enhance concentration sensitivity is needed for capillary electrophoresis (CE) to become a practical method to analyze trace levels of analytes in real samples. In this report, the development of a novel on-line preconcentration technique combining dynamic pH junction and sweeping modes of focusing is applied to the sensitive and selective analysis of three flavin derivatives: riboflavin, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD). Picomolar (pM) detectability of flavins by CE with laser-induced fluorescence (LIF) detection is demonstrated through effective focusing of large sample volumes (up to 22% capillary length) using a dual pH junction-sweeping focusing mode. This results in greater than a 1,200-fold improvement in sensitivity relative to conventional injection methods, giving a limit of detection (S/N = 3) of approximately 4.0 pM for FAD and FMN. Flavin focusing is examined in terms of analyte mobility dependence on buffer pH, borate complexation and SDS interaction. Dynamic pH junction-sweeping extends on-line focusing to both neutral (hydrophobic) and weakly acidic (hydrophilic) species and is considered useful in cases when either conventional sweeping or dynamic pH junction techniques used alone are less effective for certain classes of analytes. Enhanced focusing performance by this hyphenated method was demonstrated by greater than a 4-fold reduction in flavin bandwidth, as compared to either sweeping or dynamic pH junction, reflected by analyte detector bandwidths <0.20 cm. Novel on-line focusing strategies are required to improve sensitivity in CE, which may be applied toward more effective biochemical analysis methods for diverse types of analytes.

Supercontinuum Fourier transform spectrometry with balanced detection on a single photodiode

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

Goncharov, Vasily; Hall, Gregory

Here, we have developed phase-sensitive signal detection and processing algorithms for Fourier transform spectrometers fitted with supercontinuum sources for applications requiring ultimate sensitivity. Similar to well-established approach of source noise cancellation through balanced detection of monochromatic light, our method is capable of reducing the relative intensity noise of polychromatic light by 40 dB. Unlike conventional balanced detection, which relies on differential absorption measured with a well matched pair of photo-detectors, our algorithm utilizes phase-sensitive differential detection on a single photodiode and is capable of the real-time correction for instabilities in supercontinuum spectral structure over a broad range of wavelengths. Inmore » the resulting method is universal in terms of applicable wavelengths and compatible with commercial spectrometers. We present a proof-of-principle experimental« less

Supercontinuum Fourier transform spectrometry with balanced detection on a single photodiode

DOE PAGES

Goncharov, Vasily; Hall, Gregory

2016-08-25

Here, we have developed phase-sensitive signal detection and processing algorithms for Fourier transform spectrometers fitted with supercontinuum sources for applications requiring ultimate sensitivity. Similar to well-established approach of source noise cancellation through balanced detection of monochromatic light, our method is capable of reducing the relative intensity noise of polychromatic light by 40 dB. Unlike conventional balanced detection, which relies on differential absorption measured with a well matched pair of photo-detectors, our algorithm utilizes phase-sensitive differential detection on a single photodiode and is capable of the real-time correction for instabilities in supercontinuum spectral structure over a broad range of wavelengths. Inmore » the resulting method is universal in terms of applicable wavelengths and compatible with commercial spectrometers. We present a proof-of-principle experimental« less

Novel method for determination of zinc traces in beverages and water samples by solid surface fluorescence using a conventional quartz cuvette.

PubMed

Talio, María Carolina; Acosta, María Gimena; Acosta, Mariano; Olsina, Roberto; Fernández, Liliana P

2015-05-15

A new method for zinc pre-concentration/separation and determination by molecular fluorescence is proposed. The metal was complexed with o-phenanthroline and eosin at pH 7.5 in Tris; a piece of filter paper was used as a solid support and solid fluorescent emission measured using a conventional quartz cuvette. Under optimal conditions, the limits of detection and quantification were 0.36 × 10(-3) and 1.29 × 10(-3) μg L(-1), respectively, and the linear range from 1.29 × 10(-3) to 4.50 μg L(-1). This method showed good sensitivity and selectivity, and it was applied to the determination of zinc in foods and tap water. The absence of filtration reduced the consumption of water and electricity. Additionally, the use of common filter papers makes it a simpler and more rapid alternative to conventional methods, with sensitivity and accuracy similar to atomic spectroscopies using a typical laboratory instrument. Copyright © 2014 Elsevier Ltd. All rights reserved.

Double Threshold Energy Detection Based Cooperative Spectrum Sensing for Cognitive Radio Networks with QoS Guarantee

NASA Astrophysics Data System (ADS)

Hu, Hang; Yu, Hong; Zhang, Yongzhi

2013-03-01

Cooperative spectrum sensing, which can greatly improve the ability of discovering the spectrum opportunities, is regarded as an enabling mechanism for cognitive radio (CR) networks. In this paper, we employ a double threshold detection method in energy detector to perform spectrum sensing, only the CR users with reliable sensing information are allowed to transmit one bit local decision to the fusion center. Simulation results will show that our proposed double threshold detection method could not only improve the sensing performance but also save the bandwidth of the reporting channel compared with the conventional detection method with one threshold. By weighting the sensing performance and the consumption of system resources in a utility function that is maximized with respect to the number of CR users, it has been shown that the optimal number of CR users is related to the price of these Quality-of-Service (QoS) requirements.

Affinity Biosensors for Detection of Mycotoxins in Food.

PubMed

Evtugyn, Gennady; Subjakova, Veronika; Melikishvili, Sopio; Hianik, Tibor

2018-01-01

This chapter reviews recent achievements in methods of detection of mycotoxins in food. Special focus is on the biosensor technology that utilizes antibodies and nucleic acid aptamers as receptors. Development of biosensors is based on the immobilization of antibodies or aptamers onto various conventional supports like gold layer, but also on nanomaterials such as graphene oxide, carbon nanotubes, and quantum dots that provide an effective platform for achieving high sensitivity of detection using various physical methods, including electrochemical, mass sensitive, and optical. The biosensors developed so far demonstrate high sensitivity typically in subnanomolar limit of detection. Several biosensors have been validated in real samples. The sensitivity of biosensors is similar and, in some cases, even better than traditional analytical methods such as ELISA or chromatography. We believe that future trends will be focused on improving biosensor properties toward practical application in food industry. © 2018 Elsevier Inc. All rights reserved.

Sensitivity of Helicobacter pylori detection by Giemsa staining is poor in comparison with immunohistochemistry and fluorescent in situ hybridization and strongly depends on inflammatory activity.

PubMed

Kocsmár, Éva; Szirtes, Ildikó; Kramer, Zsófia; Szijártó, Attila; Bene, László; Buzás, György Miklós; Kenessey, István; Bronsert, Peter; Csanadi, Agnes; Lutz, Lisa; Werner, Martin; Wellner, Ulrich Friedrich; Kiss, András; Schaff, Zsuzsa; Lotz, Gábor

2017-08-01

Conventional stainings (including H&E and special stains like Giemsa) are the most widely applied histopathologic detection methods of Helicobacter pylori (HP). We aimed to compare the diagnostic performance of Giemsa staining with immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH) on a monocentric cohort of 2896 gastric biopsies and relate results to histologic alterations in order to find such histopathologic subgroups in which these methods underperform. All cases were categorized regarding presence or absence of chronic gastritis, inflammatory activity, and mucosal structural alterations. Giemsa revealed 687 cases (23.7%), IHC 795 cases (27.5%), and FISH 788 cases (27.2%) as being HP positive. Giemsa showed significantly lower overall sensitivity (83.3%) compared to IHC (98.8%) and FISH (98.0%). Moreover, the sensitivity of Giemsa dramatically dropped to 33.6% in the nonactive cases. We found that sensitivity of Giemsa strongly depends on HP density and, accordingly, on the presence of activity. Structural alterations (intestinal metaplasia, atrophy, etc.) had only no or weak effect on sensitivity of the three stainings. Both IHC and FISH proved to be equally reliable HP detecting techniques whose diagnostic performance is minimally influenced by mucosal inflammatory and structural alterations contrary to conventional stainings. We highly recommend immunohistochemistry for clinically susceptible, nonactive chronic gastritis cases, if the conventional stain-based HP detection is negative. Moreover, we recommend to use IHC more widely as basic HP stain. Helicobacter pylori FISH technique is primarily recommended to determine bacterial clarithromycin resistance. Furthermore, it is another accurate diagnostic tool for HP. © 2017 John Wiley & Sons Ltd.

Improving flow patterns and spillage characteristics of a box-type commercial kitchen hood.

PubMed

Huang, Rong Fung; Chen, Jia-Kun; Han, Meng-Ji; Priyambodo, Yusuf

2014-01-01

A conventional box-type commercial kitchen hood and its improved version (termed the "IQV commercial kitchen hood") were studied using the laser-assisted smoke flow visualization technique and tracer-gas (sulfur hexafluoride) detection methods. The laser-assisted smoke flow visualization technique qualitatively revealed the flow field of the hood and the areas apt for leakages of hood containment. The tracer-gas concentration detection method measured the quantitative leakage levels of the hood containment. The oil mists that were generated in the conventional box-type commercial kitchen hood leaked significantly into the environment from the areas near the front edges of ceiling and side walls. Around these areas, the boundary-layer separation occurred, inducing highly unsteady and turbulent recirculating flow, and leading to spillages of hood containment due to inappropriate aerodynamic design at the front edges of the ceiling and side walls. The tracer-gas concentration measurements on the conventional box-type commercial kitchen hood showed that the sulfur hexafluoride concentrations detected at the hood face attained very large values on an order of magnitude about 10(3)-10(4) ppb. By combining the backward-offset narrow suction slot, deflection plates, and quarter-circular arcs at the hood entrance, the IQV commercial kitchen hood presented a flow field containing four backward-inclined cyclone flow structures. The oil mists generated by cooking were coherently confined in these upward-rising cyclone flow structures and finally exhausted through the narrow suction slot. The tracer-gas concentration measurements on the IQV commercial kitchen hood showed that the order of magnitude of the sulfur hexafluoride concentrations detected at the hood face is negligibly small--only about 10(0) ppb across the whole hood face.

Geologic Carbon Sequestration Leakage Detection: A Physics-Guided Machine Learning Approach

NASA Astrophysics Data System (ADS)

Lin, Y.; Harp, D. R.; Chen, B.; Pawar, R.

2017-12-01

One of the risks of large-scale geologic carbon sequestration is the potential migration of fluids out of the storage formations. Accurate and fast detection of this fluids migration is not only important but also challenging, due to the large subsurface uncertainty and complex governing physics. Traditional leakage detection and monitoring techniques rely on geophysical observations including pressure. However, the resulting accuracy of these methods is limited because of indirect information they provide requiring expert interpretation, therefore yielding in-accurate estimates of leakage rates and locations. In this work, we develop a novel machine-learning technique based on support vector regression to effectively and efficiently predict the leakage locations and leakage rates based on limited number of pressure observations. Compared to the conventional data-driven approaches, which can be usually seem as a "black box" procedure, we develop a physics-guided machine learning method to incorporate the governing physics into the learning procedure. To validate the performance of our proposed leakage detection method, we employ our method to both 2D and 3D synthetic subsurface models. Our novel CO2 leakage detection method has shown high detection accuracy in the example problems.

Application of tetraplex PCR for detection of Vibrio cholerae, V. parahaemolyticus, V. vulnificus and V. mimicus in cockle.

PubMed

Senachai, Pachara; Chomvarin, Chariya; Namwat, Wises; Wongboot, Warawan; Wongwajana, Suwin; Tangkanakul, Waraluk

2013-03-01

A tetraplex PCR method was developed for simultaneous detection of Vibrio cholerae, V. parahaemolyticus, V. vulnificus and V. mimicus in cockle samples in comparison with conventional culture method. Specific primers targeting ompW of V. cholerae, tl of V. parahaemolyticus, hsp60 of V. vulnificus and sodB of V. mimicus were employed in the same PCR. Detection limit of the tetraplex PCR assay was 104 cfu/ml (400 cfu/PCR reaction) for pure cultures of all four species of Vibrio. In Vibrio spiked cockle samples, the limit of detection after 6 hours enrichment in alkaline peptone water was 1 cfu/10 g of cockle tissue for three Vibrio spp, except for V. mimicus that was 102 cfu/10 g of cockle tissue. When the tetraplex PCR and culture methods were applied to 100 cockle samples, V. parahaemolyticus, V. vulnificus, V. cholerae and V. mimicus were detected in 100, 98, 80 and 9% of the samples by tetraplex PCR and in 76, 42, 0 and 0% by the culture method, respectively. This developed tetraplex PCR method should be suitable for simultaneous and rapid detection of Vibrio species in food samples and for food safety assessment.

A method of multiplex PCR for detection of field released Beauveria bassiana, a fungal entomopathogen applied for pest management in jute (Corchorus olitorius).

PubMed

Biswas, Chinmay; Dey, Piyali; Gotyal, B S; Satpathy, Subrata

2015-04-01

The fungal entomopathogen Beauveria bassiana is a promising biocontrol agent for many pests. Some B. bassiana strains have been found effective against jute pests. To monitor the survival of field released B. bassiana a rapid and efficient detection technique is essential. Conventional methods such as plating method or direct culture method which are based on cultivation on selective media followed by microscopy are time consuming and not so sensitive. PCR based methods are rapid, sensitive and reliable. A single primer PCR may fail to amplify some of the strains. However, multiplex PCR increases the possibility of detection as it uses multiple primers. Therefore, in the present investigation a multiplex PCR protocol was developed by multiplexing three primers SCA 14, SCA 15 and SCB 9 to detect field released B. bassiana strains from soil as well as foliage of jute field. Using our multiplex PCR protocol all the five B. bassiana strains could be detected from soil and three strains viz., ITCC 6063, ITCC 4563 and ITCC 4796 could be detected even from the crop foliage after 45 days of spray.

Colorimetric Method of Loop-Mediated Isothermal Amplification with the Pre-Addition of Calcein for Detecting Flavobacterium columnare and its Assessment in Tilapia Farms.

PubMed

Suebsing, Rungkarn; Kampeera, Jantana; Sirithammajak, Sarawut; Withyachumnarnkul, Boonsirm; Turner, Warren; Kiatpathomchai, Wansika

2015-03-01

Flavobacterium columnare, the causative agent of columnaris disease in fish, affects many economically important freshwater fish species. A colorimetric method of loop-mediated isothermal amplification with the pre-addition of calcein (LAMP-calcein) was developed and used to detect the presence of F. columnare in farmed tilapia (Nile Tilapia Oreochromis niloticus and red tilapia [Nile Tilapia × Mozambique Tilapia O. mossambicus]) and rearing water. The detection method, based on a change in color from orange to green, could be performed within 45 min at 63°C. The method was highly specific, as it had no cross-detections with 14 other bacterial species, including other fish pathogens and two Flavobacterium species. The method has a minimum detection limit of 2.2 × 10(2) F. columnare CFU; thus, it is about 10 times more sensitive than conventional PCR. With this method, F. columnare was detected in gonad, gill, and blood samples from apparently healthy tilapia broodstock as well as in samples of fertilized eggs, newly hatched fry, and rearing water. The bacteria isolated from the blood were further characterized biochemically and found to be phenotypically identical to F. columnare. The amplified products from the LAMP-calcein method had 97% homology with the DNA sequence of F. columnare.

Simultaneous Detection of Genetically Modified Organisms in a Mixture by Multiplex PCR-Chip Capillary Electrophoresis.

PubMed

Patwardhan, Supriya; Dasari, Srikanth; Bhagavatula, Krishna; Mueller, Steffen; Deepak, Saligrama Adavigowda; Ghosh, Sudip; Basak, Sanjay

2015-01-01

An efficient PCR-based method to trace genetically modified food and feed products is in demand due to regulatory requirements and contaminant issues in India. However, post-PCR detection with conventional methods has limited sensitivity in amplicon separation that is crucial in multiplexing. The study aimed to develop a sensitive post-PCR detection method by using PCR-chip capillary electrophoresis (PCR-CCE) to detect and identify specific genetically modified organisms in their genomic DNA mixture by targeting event-specific nucleotide sequences. Using the PCR-CCE approach, novel multiplex methods were developed to detect MON531 cotton, EH 92-527-1 potato, Bt176 maize, GT73 canola, or GA21 maize simultaneously when their genomic DNAs in mixtures were amplified using their primer mixture. The repeatability RSD (RSDr) of the peak migration time was 0.06 and 3.88% for the MON531 and Bt176, respectively. The RSD (RSDR) of the Cry1Ac peak ranged from 0.12 to 0.40% in multiplex methods. The method was sensitive in resolving amplicon of size difference up to 4 bp. The PCR-CCE method is suitable to detect multiple genetically modified events in a composite DNA sample by tagging their event specific sequences.

Quantitative photoacoustic microscopy of optical absorption coefficients from acoustic spectra in the optical diffusive regime

NASA Astrophysics Data System (ADS)

Guo, Zijian; Favazza, Christopher; Garcia-Uribe, Alejandro; Wang, Lihong V.

2012-06-01

Photoacoustic (PA) microscopy (PAM) can image optical absorption contrast with ultrasonic spatial resolution in the optical diffusive regime. Conventionally, accurate quantification in PAM requires knowledge of the optical fluence attenuation, acoustic pressure attenuation, and detection bandwidth. We circumvent this requirement by quantifying the optical absorption coefficients from the acoustic spectra of PA signals acquired at multiple optical wavelengths. With the acoustic spectral method, the absorption coefficients of an oxygenated bovine blood phantom at 560, 565, 570, and 575 nm were quantified with errors of <3%. We also quantified the total hemoglobin concentration and hemoglobin oxygen saturation in a live mouse. Compared with the conventional amplitude method, the acoustic spectral method provides greater quantification accuracy in the optical diffusive regime. The limitations of the acoustic spectral method was also discussed.

Quantitative photoacoustic microscopy of optical absorption coefficients from acoustic spectra in the optical diffusive regime

PubMed Central

Guo, Zijian; Favazza, Christopher; Garcia-Uribe, Alejandro

2012-01-01

Abstract. Photoacoustic (PA) microscopy (PAM) can image optical absorption contrast with ultrasonic spatial resolution in the optical diffusive regime. Conventionally, accurate quantification in PAM requires knowledge of the optical fluence attenuation, acoustic pressure attenuation, and detection bandwidth. We circumvent this requirement by quantifying the optical absorption coefficients from the acoustic spectra of PA signals acquired at multiple optical wavelengths. With the acoustic spectral method, the absorption coefficients of an oxygenated bovine blood phantom at 560, 565, 570, and 575 nm were quantified with errors of <3%. We also quantified the total hemoglobin concentration and hemoglobin oxygen saturation in a live mouse. Compared with the conventional amplitude method, the acoustic spectral method provides greater quantification accuracy in the optical diffusive regime. The limitations of the acoustic spectral method was also discussed. PMID:22734767

Quantitative photoacoustic microscopy of optical absorption coefficients from acoustic spectra in the optical diffusive regime.

PubMed

Guo, Zijian; Favazza, Christopher; Garcia-Uribe, Alejandro; Wang, Lihong V

2012-06-01

Photoacoustic (PA) microscopy (PAM) can image optical absorption contrast with ultrasonic spatial resolution in the optical diffusive regime. Conventionally, accurate quantification in PAM requires knowledge of the optical fluence attenuation, acoustic pressure attenuation, and detection bandwidth. We circumvent this requirement by quantifying the optical absorption coefficients from the acoustic spectra of PA signals acquired at multiple optical wavelengths. With the acoustic spectral method, the absorption coefficients of an oxygenated bovine blood phantom at 560, 565, 570, and 575 nm were quantified with errors of <3%. We also quantified the total hemoglobin concentration and hemoglobin oxygen saturation in a live mouse. Compared with the conventional amplitude method, the acoustic spectral method provides greater quantification accuracy in the optical diffusive regime. The limitations of the acoustic spectral method was also discussed.

Classification of corn kernels contaminated with aflatoxins using fluorescence and reflectance hyperspectral image analysis

USDA-ARS?s Scientific Manuscript database

Aflatoxins are secondary metabolites produced by certain fungal species of the Aspergillus genus. Aflatoxin contamination remains a problem in agricultural products due to its toxic and carcinogenic properties. Conventional chemical methods for aflatoxin detection are time-consuming and destructive....

Analysis of spatiotemporal metabolomic dynamics for sensitively monitoring biological alterations in cisplatin-induced acute kidney injury.

PubMed

Irie, Miho; Hayakawa, Eisuke; Fujimura, Yoshinori; Honda, Youhei; Setoyama, Daiki; Wariishi, Hiroyuki; Hyodo, Fuminori; Miura, Daisuke

2018-01-29

Clinical application of the major anticancer drug, cisplatin, is limited by severe side effects, especially acute kidney injury (AKI) caused by nephrotoxicity. The detailed metabolic mechanism is still largely unknown. Here, we used an integrated technique combining mass spectrometry imaging (MSI) and liquid chromatography-mass spectrometry (LC-MS) to visualize the diverse spatiotemporal metabolic dynamics in the mouse kidney after cisplatin dosing. Biological responses to cisplatin was more sensitively detected within 24 h as a metabolic alteration, which is much earlier than possible with the conventional clinical chemistry method of blood urea nitrogen (BUN) measurement. Region-specific changes (e.g., medulla and cortex) in metabolites related to DNA damage and energy generation were observed over the 72-h exposure period. Therefore, this metabolomics approach may become a novel strategy for elucidating early renal responses to cisplatin, prior to the detection of kidney damage evaluated by conventional method. Copyright © 2018. Published by Elsevier Inc.

Determination of toxigenic fungi and aflatoxins in nuts and dried fruits using imaging and spectroscopic techniques.

PubMed

Wu, Qifang; Xie, Lijuan; Xu, Huirong

2018-06-30

Nuts and dried fruits contain rich nutrients and are thus highly vulnerable to contamination with toxigenic fungi and aflatoxins because of poor weather, processing and storage conditions. Imaging and spectroscopic techniques have proven to be potential alternative tools to wet chemistry methods for efficient and non-destructive determination of contamination with fungi and toxins. Thus, this review provides an overview of the current developments and applications in frequently used food safety testing techniques, including near infrared spectroscopy (NIRS), mid-infrared spectroscopy (MIRS), conventional imaging techniques (colour imaging (CI) and hyperspectral imaging (HSI)), and fluorescence spectroscopy and imaging (FS/FI). Interesting classification and determination results can be found in both static and on/in-line real-time detection for contaminated nuts and dried fruits. Although these techniques offer many benefits over conventional methods, challenges remain in terms of heterogeneous distribution of toxins, background constituent interference, model robustness, detection limits, sorting efficiency, as well as instrument development. Copyright © 2018 Elsevier Ltd. All rights reserved.

Effects of organic and conventional cultivation methods on composition of eggplant fruits.

PubMed

Raigón, María D; Rodríguez-Burruezo, Adrián; Prohens, Jaime

2010-06-09

Organic food is associated by the general public with improved nutritional properties, and this has led to increasing demand for organic vegetables. The effects of organic and conventional cultivation methods on dry matter, protein, minerals, and total phenolic content has been studied for two successive years in two landraces and one commercial hybrid of eggplant. In the first year, organically produced eggplants had higher mean contents (expressed on a fresh weight basis) of K (196 vs 171 mg 100 g(-1)), Ca (11.1 vs 8.7 mg 100 g(-1)), Mg (6.0 vs 4.6 mg 100 g(-1)), and total phenolics (49.8 vs 38.2 mg 100 g(-1)) than conventionally grown eggplants. In the second year, in which matched plots having a history of organic management were cultivated following organic or conventional fertilization practices, organically produced eggplants still had higher contents of K (272 vs 249 mg 100 g(-1)) and Mg (8.8 vs 7.6), as well as of Cu (0.079 vs 0.065 mg 100 g(-1)), than conventionally fertilized eggplants. Conventionally cultivated eggplants had a higher polyphenol oxidase activity than organically cultivated ones (3.19 vs 2.17 enzyme activity units), although no differences in browning were observed. Important differences in mineral concentrations between years were detected, which resulted in many correlations among mineral contents being significant. The first component of the principal component analysis separates the eggplants according to year, whereas the second component separates them according to the cultivation method (organic or conventional). Overall, the results show that organic management and fertilization have a positive effect on the accumulation of certain beneficial minerals and phenolic compounds in eggplant and that organically and conventionally produced eggplants might be distinguished according to their composition profiles.

Comparison between nasopharyngeal swab and nasal wash, using culture and PCR, in the detection of potential respiratory pathogens.

PubMed

Gritzfeld, Jenna F; Roberts, Paul; Roche, Lorna; El Batrawy, Sherouk; Gordon, Stephen B

2011-04-13

Nasopharyngeal carriage of potential pathogens is important as it is both the major source of transmission and the prerequisite of invasive disease. New methods for detecting carriage could improve comfort, accuracy and laboratory utility. The aims of this study were to compare the sensitivities of a nasopharyngeal swab (NPS) and a nasal wash (NW) in detecting potential respiratory pathogens in healthy adults using microbiological culture and PCR. Healthy volunteers attended for nasal washing and brushing of the posterior nasopharynx. Conventional and real-time PCR were used to detect pneumococcus and meningococcus. Statistical differences between the two nasal sampling methods were determined using a nonparametric Mann-Whitney U test; differences between culture and PCR methods were determined using the McNemar test.Nasal washing was more comfortable for volunteers than swabbing (n = 24). In detection by culture, the NW was significantly more likely to detect pathogens than the NPS (p < 0.00001). Overall, there was a low carriage rate of pathogens in this sample; no significant difference was seen in the detection of bacteria between culture and PCR methods. Nasal washing and PCR may provide effective alternatives to nasopharyngeal swabbing and classical microbiology, respectively.

Molecular Methods for the Detection of Mycoplasma and Ureaplasma Infections in Humans

PubMed Central

Waites, Ken B.; Xiao, Li; Paralanov, Vanya; Viscardi, Rose M.; Glass, John I.

2012-01-01

Mycoplasma and Ureaplasma species are well-known human pathogens responsible for a broad array of inflammatory conditions involving the respiratory and urogenital tracts of neonates, children, and adults. Greater attention is being given to these organisms in diagnostic microbiology, largely as a result of improved methods for their laboratory detection, made possible by powerful molecular-based techniques that can be used for primary detection in clinical specimens. For slow-growing species, such as Mycoplasma pneumoniae and Mycoplasma genitalium, molecular-based detection is the only practical means for rapid microbiological diagnosis. Most molecular-based methods used for detection and characterization of conventional bacteria have been applied to these organisms. A complete genome sequence is available for one or more strains of all of the important human pathogens in the Mycoplasma and Ureaplasma genera. Information gained from genome analyses and improvements in efficiency of DNA sequencing are expected to significantly advance the field of molecular detection and genotyping during the next few years. This review provides a summary and critical review of methods suitable for detection and characterization of mycoplasmas and ureaplasmas of humans, with emphasis on molecular genotypic techniques. PMID:22819362

Role of conventional radiology and MRi defecography of pelvic floor hernias

PubMed Central

2013-01-01

Background Purpose of the study is to define the role of conventional radiology and MRI in the evaluation of pelvic floor hernias in female pelvic floor disorders. Methods A MEDLINE and PubMed search was performed for journals before March 2013 with MeSH major terms 'MR Defecography' and 'pelvic floor hernias'. Results The prevalence of pelvic floor hernias at conventional radiology was higher if compared with that at MRI. Concerning the hernia content, there were significantly more enteroceles and sigmoidoceles on conventional radiology than on MRI, whereas, in relation to the hernia development modalities, the prevalence of elytroceles, edroceles, and Douglas' hernias at conventional radiology was significantly higher than that at MRI. Conclusions MRI shows lower sensitivity than conventional radiology in the detection of pelvic floor hernias development. The less-invasive MRI may have a role in a better evaluation of the entire pelvic anatomy and pelvic organ interaction especially in patients with multicompartmental defects, planned for surgery. PMID:24267789

Unconstrained Respiration Measurement and Respiratory Arrest Detection Method by Dynamic Threshold in Transferring Patients by Stretchers

NASA Astrophysics Data System (ADS)

Kurihara, Yosuke; Watanabe, Kajiro; Kobayashi, Kazuyuki; Tanaka, Hiroshi

General anesthesia used for surgical operations may cause unstable conditions of the patients after the operations, which could lead to respiratory arrests. Under such circumstances, nurses could fail in finding the change of the conditions, and other malpractices could also occur. It is highly possible that such malpractices may occur while transferring a patient from ICU to the room using a stretcher. Monitoring the change in the blood oxygen saturation concentration and other vital signs to detect a respiratory arrest is not easy when transferring a patient on a stretcher. Here we present several noise reduction system and algorithm to detect respiratory arrests in transferring a patient, based on the unconstrained air pressure method that the authors presented previously. As the result, when the acceleration level of the stretcher noise was 0.5G, the respiratory arrest detection ratio using this novel method was 65%, while that with the conventional method was 0%.

Evaluation of selective control information detection scheme in orthogonal frequency division multiplexing-based radio-over-fiber and visible light communication links

NASA Astrophysics Data System (ADS)

Dalarmelina, Carlos A.; Adegbite, Saheed A.; Pereira, Esequiel da V.; Nunes, Reginaldo B.; Rocha, Helder R. O.; Segatto, Marcelo E. V.; Silva, Jair A. L.

2017-05-01

Block-level detection is required to decode what may be classified as selective control information (SCI) such as control format indicator in 4G-long-term evolution systems. Using optical orthogonal frequency division multiplexing over radio-over-fiber (RoF) links, we report the experimental evaluation of an SCI detection scheme based on a time-domain correlation (TDC) technique in comparison with the conventional maximum likelihood (ML) approach. When compared with the ML method, it is shown that the TDC method improves detection performance over both 20 and 40 km of standard single mode fiber (SSMF) links. We also report a performance analysis of the TDC scheme in noisy visible light communication channel models after propagation through 40 km of SSMF. Experimental and simulation results confirm that the TDC method is attractive for practical orthogonal frequency division multiplexing-based RoF and fiber-wireless systems. Unlike the ML method, another key benefit of the TDC is that it requires no channel estimation.

A vision-based fall detection algorithm of human in indoor environment

NASA Astrophysics Data System (ADS)

Liu, Hao; Guo, Yongcai

2017-02-01

Elderly care becomes more and more prominent in China as the population is aging fast and the number of aging population is large. Falls, as one of the biggest challenges in elderly guardianship system, have a serious impact on both physical health and mental health of the aged. Based on feature descriptors, such as aspect ratio of human silhouette, velocity of mass center, moving distance of head and angle of the ultimate posture, a novel vision-based fall detection method was proposed in this paper. A fast median method of background modeling with three frames was also suggested. Compared with the conventional bounding box and ellipse method, the novel fall detection technique is not only applicable for recognizing the fall behaviors end of lying down but also suitable for detecting the fall behaviors end of kneeling down and sitting down. In addition, numerous experiment results showed that the method had a good performance in recognition accuracy on the premise of not adding the cost of time.

Eukaryote-Made Thermostable DNA Polymerase Enables Rapid PCR-Based Detection of Mycoplasma, Ureaplasma and Other Bacteria in the Amniotic Fluid of Preterm Labor Cases

PubMed Central

Yoneda, Noriko; Yoneda, Satoshi; Mori, Masashi; Tabata, Homare; Minami, Hiroshi; Saito, Shigeru; Kitajima, Isao

2015-01-01

Background Intra-amniotic infection has long been recognized as the leading cause of preterm delivery. Microbial culture is the gold standard for the detection of intra-amniotic infection, but several days are required, and many bacterial species in the amniotic fluid are difficult to cultivate. Methods We developed a novel nested-PCR-based assay for detecting Mycoplasma, Ureaplasma, other bacteria and fungi in amniotic fluid samples within three hours of sample collection. To detect prokaryotes, eukaryote-made thermostable DNA polymerase, which is free from bacterial DNA contamination, is used in combination with bacterial universal primers. In contrast, to detect eukaryotes, conventional bacterially-made thermostable DNA polymerase is used in combination with fungal universal primers. To assess the validity of the PCR assay, we compared the PCR and conventional culture results using 300 amniotic fluid samples. Results Based on the detection level (positive and negative), 93.3% (280/300) of Mycoplasma, 94.3% (283/300) of Ureaplasma, 89.3% (268/300) of other bacteria and 99.7% (299/300) of fungi matched the culture results. Meanwhile, concerning the detection of bacteria other than Mycoplasma and Ureaplasma, 228 samples were negative according to the PCR method, 98.2% (224/228) of which were also negative based on the culture method. Employing the devised primer sets, mixed amniotic fluid infections of Mycoplasma, Ureaplasma and/or other bacteria could be clearly distinguished. In addition, we also attempted to compare the relative abundance in 28 amniotic fluid samples with mixed infection, and judged dominance by comparing the Ct values of quantitative real-time PCR. Conclusions We developed a novel PCR assay for the rapid detection of Mycoplasma, Ureaplasma, other bacteria and fungi in amniotic fluid samples. This assay can also be applied to accurately diagnose the absence of bacteria in samples. We believe that this assay will positively contribute to the treatment of intra-amniotic infection and the prevention of preterm delivery. PMID:26042418

Sensitive detection of dopamine via leucodopaminechrome on polyacrylic acid-coated ceria nanorods

NASA Astrophysics Data System (ADS)

Sheng, Weiqin; Zheng, Liang; Liu, Yan; Zhao, Xueqin; Weng, Jian; Zhang, Yang

2017-09-01

The major hurdle in detection of dopamine (DA) by electro-analysis is the presence of physiological interferents with a similar oxidation potential of DA. The conventional method is to enlarge the difference of their oxidation potentials. Here, we report an unconventional method to detect DA via leucodopaminechrome on CeO2 nanorods. Leucodopaminechrome is produced from the cyclization of dopamine-quinone, a product of two-electron oxidation of DA. Thus, its concentration is proportional to the DA concentration. Determining DA is demonstrated by measuring the reduction current of leucodopaminechrome on CeO2 nanorods. CeO2 nanorods demonstrate high electrocatalytic activity for reduction of leucodopaminechrome with a low potential at -0.27 V. The low detection potential of leucodopaminechrome can avoid the interference from ascorbic acid (AA) and uric acid (UA). Therefore, detecting DA via leucodopaminechrome is an effective method to avoid interference from AA and UA, and the suggested biosensor also displays good reproducibility and stability.

Generation and coherent detection of QPSK signal using a novel method of digital signal processing

NASA Astrophysics Data System (ADS)

Zhao, Yuan; Hu, Bingliang; He, Zhen-An; Xie, Wenjia; Gao, Xiaohui

2018-02-01

We demonstrate an optical quadrature phase-shift keying (QPSK) signal transmitter and an optical receiver for demodulating optical QPSK signal with homodyne detection and digital signal processing (DSP). DSP on the homodyne detection scheme is employed without locking the phase of the local oscillator (LO). In this paper, we present an extracting one-dimensional array of down-sampling method for reducing unwanted samples of constellation diagram measurement. Such a novel scheme embodies the following major advantages over the other conventional optical QPSK signal detection methods. First, this homodyne detection scheme does not need strict requirement on LO in comparison with linear optical sampling, such as having a flat spectral density and phase over the spectral support of the source under test. Second, the LabVIEW software is directly used for recovering the QPSK signal constellation without employing complex DSP circuit. Third, this scheme is applicable to multilevel modulation formats such as M-ary PSK and quadrature amplitude modulation (QAM) or higher speed signals by making minor changes.

Interdigitated microelectrode based impedance biosensor for detection of salmonella enteritidis in food samples

NASA Astrophysics Data System (ADS)

Kim, G.; Morgan, M.; Hahm, B. K.; Bhunia, A.; Mun, J. H.; Om, A. S.

2008-03-01

Salmonella enteritidis outbreaks continue to occur, and S. enteritidis-related outbreaks from various food sources have increased public awareness of this pathogen. Conventional methods for pathogens detection and identification are labor-intensive and take days to complete. Some immunological rapid assays are developed, but these assays still require prolonged enrichment steps. Recently developed biosensors have shown great potential for the rapid detection of foodborne pathogens. To develop the biosensor, an interdigitated microelectrode (IME) was fabricated by using semiconductor fabrication process. Anti-Salmonella antibodies were immobilized based on avidin-biotin binding on the surface of the IME to form an active sensing layer. To increase the sensitivity of the sensor, three types of sensors that have different electrode gap sizes (2 μm, 5 μm, 10 μm) were fabricated and tested. The impedimetric biosensor could detect 103 CFU/mL of Salmonella in pork meat extract with an incubation time of 5 minutes. This method may provide a simple, rapid and sensitive method to detect foodborne pathogens.

Reversed-phase HPLC analysis of levetiracetam in tablets using monolithic and conventional C18 silica columns.

PubMed

Can, Nafiz O; Arli, Goksel

2010-01-01

Development and validation of an RP-HPLC method for determination of levetiracetam in pharmaceutical tablets is described. The separation and quantification of levetiracetam and caffeine (internal standard) were performed using a single analytical procedure with two different types of stationary phases, conventional Phenomenex Gemini C18 (100 x 4.6 mm, 5 microm) and Merck Chromolith Performance RP18e (100 x 4.6 mm, macropore size 2 mm, micropore size 13 nm) monolithic silica. Five-microliter aliquots of samples were injected into the system and eluted using water-acetonitrile (90 + 10, v/v) mobile phase pumped at the rate of 1 mL/min. The analyte peaks were detected at 200 nm using a diode array detector with adequate resolution. Validation studies were performed using the method recommended by the International Conference on Harmonization, the U.S. Pharmacopeia, and AOAC INTERNATIONAL, which includes accuracy, precision, range, limits, robustness, and system suitability parameters. Levetiracetam and caffeine were detected in about 7 min using the conventional column, whereas less than 5 min was required when the monolithic column was used. Calibration plots had r values close to unity in the range of 0.8-8.0 microg/mL. Assay of levetiracetam in a tablet formulation was demonstrated as an application to real samples.

Neural adaptive observer-based sensor and actuator fault detection in nonlinear systems: Application in UAV.

PubMed

Abbaspour, Alireza; Aboutalebi, Payam; Yen, Kang K; Sargolzaei, Arman

2017-03-01

A new online detection strategy is developed to detect faults in sensors and actuators of unmanned aerial vehicle (UAV) systems. In this design, the weighting parameters of the Neural Network (NN) are updated by using the Extended Kalman Filter (EKF). Online adaptation of these weighting parameters helps to detect abrupt, intermittent, and incipient faults accurately. We apply the proposed fault detection system to a nonlinear dynamic model of the WVU YF-22 unmanned aircraft for its evaluation. The simulation results show that the new method has better performance in comparison with conventional recurrent neural network-based fault detection strategies. Copyright © 2016 ISA. Published by Elsevier Ltd. All rights reserved.

Development and Validation of Monoclonal Antibody-Based Antigen Capture ELISA for Detection of Group A Porcine Rotavirus.

PubMed

Memon, Atta Muhammad; Bhuyan, Anjuman Ara; Chen, Fangzhou; Guo, Xiaozhen; Menghwar, Harish; Zhu, Yinxing; Ku, Xugang; Chen, Shuhua; Li, Zhonghua; He, Qigai

2017-05-01

Porcine rotavirus-A (PoRVA) is one of the common causes of mild to severe dehydrating diarrhea, leading to losses in weaning and postweaning piglets. A rapid, highly specific, and sensitive antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed for detection of PoRVA, by using VP6 (a highly conserved and antigenic protein of group-A rotavirus)-directed rabbit polyclonal antibodies (capture antibody) and murine monoclonal antibodies (detector antibody). The detection limit of AC-ELISA was found to be equal to that of conventional reverse transcription-polymerase chain reaction (RT-PCR; about 10 2.5 TCID 50 /mL). For validation of the in-house AC-ELISA, 295 porcine fecal/diarrhea samples, collected from different provinces of China, were evaluated and compared with conventional RT-PCR and TaqMan RT-quantitative PCR (qPCR). The sensitivity and specificity of this in-house AC-ELISA relative to RT-qPCR were found to be 91.67% and 100%, respectively, with the strong agreement (kappa = 0.972) between these two techniques. Total detection rate with AC-ELISA, conventional RT-PCR, and RT-qPCR were found to be 11.2%, 11.5%, and 12.2%, respectively, without any statistical significant difference. Moreover, AC-ELISA failed to detect any cross-reactivity with porcine epidemic diarrhea virus, transmissible gastroenteritis virus, pseudorabies virus, and porcine circovirus-2. These results suggested that our developed method was rapid, highly specific, and sensitive, which may help in large-scale surveillance, timely detection, and preventive control of rotavirus infection in porcine farms.

Target attribute-based false alarm rejection in small infrared target detection

NASA Astrophysics Data System (ADS)

Kim, Sungho

2012-11-01

Infrared search and track is an important research area in military applications. Although there are a lot of works on small infrared target detection methods, we cannot apply them in real field due to high false alarm rate caused by clutters. This paper presents a novel target attribute extraction and machine learning-based target discrimination method. Eight kinds of target features are extracted and analyzed statistically. Learning-based classifiers such as SVM and Adaboost are developed and compared with conventional classifiers for real infrared images. In addition, the generalization capability is also inspected for various infrared clutters.

[Assessment of cervical intraepithelial neoplasia (CIN) lesions by DNA image cytometry].

PubMed

Sun, Xiao-rong; Che, Dong-yuan; Tu, Hong-zhang; Li, Dan; Wang, Jian

2006-11-01

To compare the value of conventional cytology and DNA image cytometry (DNA-ICM) assisted cytology in detection and prognostic assessment of cervical CIN lesions. 87 women were enrolled in this study. Cervical samples were collected employing cervix brushes which were then washed in Sedfix. After preparing single cell suspensions by mechanical procedure, cell monolayers were prepared by cyto-spinning the cells onto microscope slides. Two slides were prepared from each case: one slide was stained by Papanicolou staining for conventional cytology, another was stained by Feulgen-Thionin method for measurements of the amount of DNA in the cell nuclei using an automated DNA imaging cytometer. Biopsies from the cervical lesions were also taken for histopathology and Ki-67 immunohistochemistry. Of the total of 20 ASCUS cases called by conventional cytology, no CIN, nor greater lesions were found. Among the 20 cases, 7 cases did not show any cells with DNA amount greater than 5c, while CIN2 lesions were found in 11 of other 13 cases that had some aneuploid cells with DNA amount greater than 5c. Of 30 LSIL cases called by conventional cytology, CIN2 lesions were detected in 3 out of 7 cases that did not contain any aneuploid cells with DNA greater than 5c, but in 22 out of the other 23 cases that contained aneuploid cells with DNA amount greater than > 5c. Of the remaining 7 cases called HSIL by conventional cytology, all case contained aneuploid cells containing DNA greater than 5c. If cytology was used to refer all cases of LSIL and HSIL to colposcopy procedure to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive value were 58.2%, 84.4%, 86.5% and 54.0%, respectively. If DNA-ICM were used and all cases having 3 or more cells with a DNA amount greater than 5c were assessed to be referred to pathology to detect potential CIN2 or greater lesions, the sensitivity, specificity, positive predictive value and negative predictive were 72.7% , 87.5%, 90.9% and 65.1%, respectively. We also compared Ki67 positive cells in these samples and found that DNA-ICM results were comparable to this biomarker method. The study demonstrated that DNA-ICM approach can be successfully used to detect significant (i.e. CIN2 or greater) lesions, and also provide a prognostic assessment of CIN lesions.

Rapid direct methods for enumeration of specific, active bacteria in water and biofilms

NASA Technical Reports Server (NTRS)

McFeters, G. A.; Pyle, B. H.; Lisle, J. T.; Broadaway, S. C.

1999-01-01

Conventional methods for detecting indicator and pathogenic bacteria in water may underestimate the actual population due to sublethal environmental injury, inability of the target bacteria to take up nutrients and other physiological factors which reduce bacterial culturability. Rapid and direct methods are needed to more accurately detect and enumerate active bacteria. Such a methodological advance would provide greater sensitivity in assessing the microbiological safety of water and food. The principle goal of this presentation is to describe novel approaches we have formulated for the rapid and simultaneous detection of bacteria plus the determination of their physiological activity in water and other environmental samples. The present version of our method involves the concentration of organisms by membrane filtration or immunomagnetic separation and combines an intracellular fluorochrome (CTC) for assessment of respiratory activity plus fluorescent-labelled antibody detection of specific bacteria. This approach has also been successfully used to demonstrate spatial and temporal heterogeneities of physiological activities in biofilms when coupled with cryosectioning. Candidate physiological stains include those capable of determining respiratory activity, membrane potential, membrane integrity, growth rate and cellular enzymatic activities. Results obtained thus far indicate that immunomagnetic separation can provide a high degree of sensitivity in the recovery of seeded target bacteria (Escherichia coli O157:H7) in water and hamburger. The captured and stained target bacteria are then enumerated by either conventional fluorescence microscopy or ChemScan(R), a new instrument that is very sensitive and rapid. The ChemScan(R) laser scanning instrument (Chemunex, Paris, France) provides the detection of individual fluorescently labelled bacterial cells using three emission channels in less than 5 min. A high degree of correlation has been demonstrated between results obtained with the ChemScan and traditional plate counts of mixed natural bacterial populations in water. The continuing evolution of these methods will be valuable in the rapid and accurate analysis of environmental samples.

A novel signal amplification technology for ELISA based on catalyzed reporter deposition. Demonstration of its applicability for measuring aflatoxin B(1).

PubMed

Bhattacharya, D; Bhattacharya, R; Dhar, T K

1999-11-19

In an earlier communication we have described a novel signal amplification technology termed Super-CARD, which is able to significantly improve antigen detection sensitivity in conventional Dot-ELISA by approximately 10(5)-fold. The method utilizes hitherto unreported synthesized electron rich proteins containing multiple phenolic groups which, when immobilized over a solid phase as blocking agent, markedly increases the signal amplification capability of the existing CARD method (Bhattacharya, R., Bhattacharya, D., Dhar, T.K., 1999. A novel signal amplification technology based on catalyzed reporter deposition and its application in a Dot-ELISA with ultra high sensitivity. J. Immunol. Methods 227, 31.). In this paper we describe the utilization of this Super-CARD amplification technique in ELISA and its applicability for the rapid determination of aflatoxin B(1) (AFB(1)) in infected seeds. Using this method under identical conditions, the increase in absorbance over the CARD method was approximately 400%. The limit of detection of AFB(1) by this method was 0.1 pg/well, the sensitivity enhancement being 5-fold over the optimized CARD ELISA. Furthermore, the total incubation time was reduced to 16 min compared to 50 min for the CARD method. Assay specificity was not adversely affected and the amount of AFB(1) measured in seed extracts correlated well with the values obtained by conventional ELISA.

Standard addition with internal standardisation as an alternative to using stable isotope labelled internal standards to correct for matrix effects-Comparison and validation using liquid chromatography-​tandem mass spectrometric assay of vitamin D.

PubMed

Hewavitharana, Amitha K; Abu Kassim, Nur Sofiah; Shaw, Paul Nicholas

2018-06-08

With mass spectrometric detection in liquid chromatography, co-eluting impurities affect the analyte response due to ion suppression/enhancement. Internal standard calibration method, using co-eluting stable isotope labelled analogue of each analyte as the internal standard, is the most appropriate technique available to correct for these matrix effects. However, this technique is not without drawbacks, proved to be expensive because separate internal standard for each analyte is required, and the labelled compounds are expensive or require synthesising. Traditionally, standard addition method has been used to overcome the matrix effects in atomic spectroscopy and was a well-established method. This paper proposes the same for mass spectrometric detection, and demonstrates that the results are comparable to those with the internal standard method using labelled analogues, for vitamin D assay. As conventional standard addition procedure does not address procedural errors, we propose the inclusion of an additional internal standard (not co-eluting). Recoveries determined on human serum samples show that the proposed method of standard addition yields more accurate results than the internal standardisation using stable isotope labelled analogues. The precision of the proposed method of standard addition is superior to the conventional standard addition method. Copyright © 2018 Elsevier B.V. All rights reserved.

Comparison of Four Human Papillomavirus Genotyping Methods: Next-generation Sequencing, INNO-LiPA, Electrochemical DNA Chip, and Nested-PCR.

PubMed

Nilyanimit, Pornjarim; Chansaenroj, Jira; Poomipak, Witthaya; Praianantathavorn, Kesmanee; Payungporn, Sunchai; Poovorawan, Yong

2018-03-01

Human papillomavirus (HPV) infection causes cervical cancer, thus necessitating early detection by screening. Rapid and accurate HPV genotyping is crucial both for the assessment of patients with HPV infection and for surveillance studies. Fifty-eight cervicovaginal samples were tested for HPV genotypes using four methods in parallel: nested-PCR followed by conventional sequencing, INNO-LiPA, electrochemical DNA chip, and next-generation sequencing (NGS). Seven HPV genotypes (16, 18, 31, 33, 45, 56, and 58) were identified by all four methods. Nineteen HPV genotypes were detected by NGS, but not by nested-PCR, INNO-LiPA, or electrochemical DNA chip. Although NGS is relatively expensive and complex, it may serve as a sensitive HPV genotyping method. Because of its highly sensitive detection of multiple HPV genotypes, NGS may serve as an alternative for diagnostic HPV genotyping in certain situations. © The Korean Society for Laboratory Medicine

Non-destructive techniques for the detection of fungal infection in cereal grains.

PubMed

Orina, Irene; Manley, Marena; Williams, Paul J

2017-10-01

Infection of cereal grains by fungi is a serious problem worldwide. Depending on the environmental conditions, cereal grains may be colonised by different species of fungi. These fungi cause reduction in yield, quality and nutritional value of the grain; and of major concern is their production of mycotoxins which are harmful to both humans and animals. Early detection of fungal contamination is an essential control measure for ensuring storage longevity and food safety. Conventional methods for detection of fungal infection, such as culture and colony techniques or immunological methods are either slow, labour intensive or difficult to automate. In recent years, there has been an increasing need to develop simple, rapid, non-destructive methods for early detection of fungal infection and mycotoxins contamination in cereal grains. Methods such as near infrared (NIR) spectroscopy, NIR hyperspectral imaging, and electronic nose were evaluated for these purposes. This paper reviews the different non-destructive techniques that have been considered thus far for detection of fungal infection and mycotoxins in cereal grains, including their principles, application and limitations. Copyright © 2017 Elsevier Ltd. All rights reserved.

ROKU: a novel method for identification of tissue-specific genes

PubMed Central

Kadota, Koji; Ye, Jiazhen; Nakai, Yuji; Terada, Tohru; Shimizu, Kentaro

2006-01-01

Background One of the important goals of microarray research is the identification of genes whose expression is considerably higher or lower in some tissues than in others. We would like to have ways of identifying such tissue-specific genes. Results We describe a method, ROKU, which selects tissue-specific patterns from gene expression data for many tissues and thousands of genes. ROKU ranks genes according to their overall tissue specificity using Shannon entropy and detects tissues specific to each gene if any exist using an outlier detection method. We evaluated the capacity for the detection of various specific expression patterns using synthetic and real data. We observed that ROKU was superior to a conventional entropy-based method in its ability to rank genes according to overall tissue specificity and to detect genes whose expression pattern are specific only to objective tissues. Conclusion ROKU is useful for the detection of various tissue-specific expression patterns. The framework is also directly applicable to the selection of diagnostic markers for molecular classification of multiple classes. PMID:16764735

CAFFEINE SPECIFICITY OF VARIOUS NON-IMPRINTED POLYMERS IN AQUEOUS MEDIA

EPA Science Inventory

Limitations exist in applying the conventional microbial methods to the detection of human fecal contamination in water. Certain organic compounds such as caffeine, have been reported by the U.S. Geological Survey as a more suitable tracer. The employment of caffeine has been h...

Efficacy of low-pressure foam cleaning compared to conventional cleaning methods in the removal of bacteria from surfaces associated with convenience food.

PubMed

Lambrechts, A A; Human, I S; Doughari, J H; Lues, J F R

2014-09-01

Food borne illnesses and food poisoning are cause for concern globally. The diseases are often caused by food contamination with pathogenic bacteria due largely to poor sanitary habits or storage conditions. Prevalence of some bacteria on cleaned and sanitised food contact surfaces from eight convenience food plants in Gauteng (South Africa) was investigated with the view to evaluate the efficacy of the cleaning methods used with such food contact surfaces. The microbial load of eight convenience food manufacturing plants was determined by sampling stainless steel food contact surfaces after they had been cleaned and sanitised at the end of a day's shift. Samples were analysed for Total Plate Count (TPC), Escherichia coli, Salmonella species, Staphylococcus aureus and Listeria species. Results showed that 59 % of the total areas sampled for TPC failed to comply with the legal requirements for surfaces, according to the Foodstuffs, Cosmetics and Disinfectants Act (< 100 cfu.cm(-2)). S. aureus and Salmonella were not detected, but Listeria was detected in 23 % and E. coli in 1.3 % of the samples. Fifty percent (50 %) of the plants applied conventional cleaning methods for cleaning and sanitation and 50 % used the low-pressure foam (LPF) method. There was significant difference (P ≤ 0.05) between the mean TPC values of the conventional cleaning method (14 358.82) compared to that of LPF method (2 386.51) but no significant difference (P > 0.05) in terms of Listeria species isolates obtained from both cleaning methods. The LPF method proved to be the superior cleaning option for lowering TPC counts. Regardless of cleaning method used, pathogens continued to flourish on various surfaces, including dry stainless steel, posing a contamination hazard for a considerable period depending on the contamination level and type of pathogen. Intensive training for proper chemical usage and strict procedural compliance among workers for efficient cleaning procedures is recommended.

Automatic adventitious respiratory sound analysis: A systematic review.

PubMed

Pramono, Renard Xaviero Adhi; Bowyer, Stuart; Rodriguez-Villegas, Esther

2017-01-01

Automatic detection or classification of adventitious sounds is useful to assist physicians in diagnosing or monitoring diseases such as asthma, Chronic Obstructive Pulmonary Disease (COPD), and pneumonia. While computerised respiratory sound analysis, specifically for the detection or classification of adventitious sounds, has recently been the focus of an increasing number of studies, a standardised approach and comparison has not been well established. To provide a review of existing algorithms for the detection or classification of adventitious respiratory sounds. This systematic review provides a complete summary of methods used in the literature to give a baseline for future works. A systematic review of English articles published between 1938 and 2016, searched using the Scopus (1938-2016) and IEEExplore (1984-2016) databases. Additional articles were further obtained by references listed in the articles found. Search terms included adventitious sound detection, adventitious sound classification, abnormal respiratory sound detection, abnormal respiratory sound classification, wheeze detection, wheeze classification, crackle detection, crackle classification, rhonchi detection, rhonchi classification, stridor detection, stridor classification, pleural rub detection, pleural rub classification, squawk detection, and squawk classification. Only articles were included that focused on adventitious sound detection or classification, based on respiratory sounds, with performance reported and sufficient information provided to be approximately repeated. Investigators extracted data about the adventitious sound type analysed, approach and level of analysis, instrumentation or data source, location of sensor, amount of data obtained, data management, features, methods, and performance achieved. A total of 77 reports from the literature were included in this review. 55 (71.43%) of the studies focused on wheeze, 40 (51.95%) on crackle, 9 (11.69%) on stridor, 9 (11.69%) on rhonchi, and 18 (23.38%) on other sounds such as pleural rub, squawk, as well as the pathology. Instrumentation used to collect data included microphones, stethoscopes, and accelerometers. Several references obtained data from online repositories or book audio CD companions. Detection or classification methods used varied from empirically determined thresholds to more complex machine learning techniques. Performance reported in the surveyed works were converted to accuracy measures for data synthesis. Direct comparison of the performance of surveyed works cannot be performed as the input data used by each was different. A standard validation method has not been established, resulting in different works using different methods and performance measure definitions. A review of the literature was performed to summarise different analysis approaches, features, and methods used for the analysis. The performance of recent studies showed a high agreement with conventional non-automatic identification. This suggests that automated adventitious sound detection or classification is a promising solution to overcome the limitations of conventional auscultation and to assist in the monitoring of relevant diseases.

Automatic adventitious respiratory sound analysis: A systematic review

PubMed Central

Bowyer, Stuart; Rodriguez-Villegas, Esther

2017-01-01

Background Automatic detection or classification of adventitious sounds is useful to assist physicians in diagnosing or monitoring diseases such as asthma, Chronic Obstructive Pulmonary Disease (COPD), and pneumonia. While computerised respiratory sound analysis, specifically for the detection or classification of adventitious sounds, has recently been the focus of an increasing number of studies, a standardised approach and comparison has not been well established. Objective To provide a review of existing algorithms for the detection or classification of adventitious respiratory sounds. This systematic review provides a complete summary of methods used in the literature to give a baseline for future works. Data sources A systematic review of English articles published between 1938 and 2016, searched using the Scopus (1938-2016) and IEEExplore (1984-2016) databases. Additional articles were further obtained by references listed in the articles found. Search terms included adventitious sound detection, adventitious sound classification, abnormal respiratory sound detection, abnormal respiratory sound classification, wheeze detection, wheeze classification, crackle detection, crackle classification, rhonchi detection, rhonchi classification, stridor detection, stridor classification, pleural rub detection, pleural rub classification, squawk detection, and squawk classification. Study selection Only articles were included that focused on adventitious sound detection or classification, based on respiratory sounds, with performance reported and sufficient information provided to be approximately repeated. Data extraction Investigators extracted data about the adventitious sound type analysed, approach and level of analysis, instrumentation or data source, location of sensor, amount of data obtained, data management, features, methods, and performance achieved. Data synthesis A total of 77 reports from the literature were included in this review. 55 (71.43%) of the studies focused on wheeze, 40 (51.95%) on crackle, 9 (11.69%) on stridor, 9 (11.69%) on rhonchi, and 18 (23.38%) on other sounds such as pleural rub, squawk, as well as the pathology. Instrumentation used to collect data included microphones, stethoscopes, and accelerometers. Several references obtained data from online repositories or book audio CD companions. Detection or classification methods used varied from empirically determined thresholds to more complex machine learning techniques. Performance reported in the surveyed works were converted to accuracy measures for data synthesis. Limitations Direct comparison of the performance of surveyed works cannot be performed as the input data used by each was different. A standard validation method has not been established, resulting in different works using different methods and performance measure definitions. Conclusion A review of the literature was performed to summarise different analysis approaches, features, and methods used for the analysis. The performance of recent studies showed a high agreement with conventional non-automatic identification. This suggests that automated adventitious sound detection or classification is a promising solution to overcome the limitations of conventional auscultation and to assist in the monitoring of relevant diseases. PMID:28552969

Identification of De Novo and Rare Inherited Copy Number Variants in Children with Syndromic Congenital Heart Defects.

PubMed

Hussein, Ibtessam R; Bader, Rima S; Chaudhary, Adeel G; Bassiouni, Randa; Alquaiti, Maha; Ashgan, Fai; Schulten, Hans-Juergen; Al Qahtani, Mohammad H

2018-06-01

Congenital heart defects (CHDs) are the most common birth defects in neonatal life. CHDs could be presented as isolated defects or associated with developmental delay (DD) and/or other congenital malformations. A small proportion of cardiac defects are caused by chromosomal abnormalities or single gene defects; however, in a large proportion of cases no genetic diagnosis could be achieved by clinical examination and conventional genetic analysis. The development of genome wide array-Comparative Genomic Hybridization technique (array-CGH) allowed for the detection of cryptic chromosomal imbalances and pathogenic copy number variants (CNVs) not detected by conventional techniques. We investigated 94 patients having CHDs associated with other malformations and/or DD. Clinical examination and Echocardiography was done to all patients to evaluate the type of CHD. To investigate for genome defects we applied high-density array-CGH 2 × 400K (41 patients) and CGH/SNP microarray 2 × 400K (Agilent) for 53 patients. Confirmation of results was done using Fluorescent in situ hybridization (FISH) or qPCR techniques in certain cases. Chromosomal abnormalities such as trisomy 18, 13, 21, microdeletions: del22q11.2, del7q11.23, del18 (p11.32; p11.21), tetrasomy 18p, trisomy 9p, del11q24-q25, add 15p, add(18)(q21.3), and der 9, 15 (q34.2; q11.2) were detected in 21/94 patients (22%) using both conventional cytogenetics methods and array-CGH technique. Cryptic chromosomal anomalies and pathogenic variants were detected in 15/73 (20.5%) cases. CNVs were observed in a large proportion of the studied samples (27/56) (48%). Clustering of variants was observed in chromosome 1p36, 1p21.1, 2q37, 3q29, 5p15, 7p22.3, 8p23, 11p15.5, 14q11.2, 15q11.2, 16p13.3, 16p11.2, 18p11, 21q22, and 22q11.2. CGH/SNP array could detect loss of heterozygosity (LOH) in different chromosomal loci in 10/25 patients. Array-CGH technique allowed for detection of cryptic chromosomal imbalances that could not be detected by conventional cytogenetics methods. CHDs associated with DD/congenital malformations presented with a relatively high rate of cryptic chromosomal abnormalities. Clustering of CNVs in certain genome loci needs further analysis to identify candidate genes that may provide clues for understanding the molecular pathway of cardiac development.

Air sampling with solid phase microextraction

NASA Astrophysics Data System (ADS)

Martos, Perry Anthony

There is an increasing need for simple yet accurate air sampling methods. The acceptance of new air sampling methods requires compatibility with conventional chromatographic equipment, and the new methods have to be environmentally friendly, simple to use, yet with equal, or better, detection limits, accuracy and precision than standard methods. Solid phase microextraction (SPME) satisfies the conditions for new air sampling methods. Analyte detection limits, accuracy and precision of analysis with SPME are typically better than with any conventional air sampling methods. Yet, air sampling with SPME requires no pumps, solvents, is re-usable, extremely simple to use, is completely compatible with current chromatographic equipment, and requires a small capital investment. The first SPME fiber coating used in this study was poly(dimethylsiloxane) (PDMS), a hydrophobic liquid film, to sample a large range of airborne hydrocarbons such as benzene and octane. Quantification without an external calibration procedure is possible with this coating. Well understood are the physical and chemical properties of this coating, which are quite similar to those of the siloxane stationary phase used in capillary columns. The log of analyte distribution coefficients for PDMS are linearly related to chromatographic retention indices and to the inverse of temperature. Therefore, the actual chromatogram from the analysis of the PDMS air sampler will yield the calibration parameters which are used to quantify unknown airborne analyte concentrations (ppb v to ppm v range). The second fiber coating used in this study was PDMS/divinyl benzene (PDMS/DVB) onto which o-(2,3,4,5,6- pentafluorobenzyl) hydroxylamine (PFBHA) was adsorbed for the on-fiber derivatization of gaseous formaldehyde (ppb v range), with and without external calibration. The oxime formed from the reaction can be detected with conventional gas chromatographic detectors. Typical grab sampling times were as small as 5 seconds. With 300 seconds sampling, the formaldehyde detection limit was 2.1 ppbv, better than any other 5 minute sampling device for formaldehyde. The first-order rate constant for product formation was used to quantify formaldehyde concentrations without a calibration curve. This spot sampler was used to sample the headspace of hair gel, particle board, plant material and coffee grounds for formaldehyde, and other carbonyl compounds, with extremely promising results. The SPME sampling devices were also used for time- weighted average sampling (30 minutes to 16 hours). Finally, the four new SPME air sampling methods were field tested with side-by-side comparisons to standard air sampling methods, showing a tremendous use of SPME as an air sampler.

A comprehensive library-based, automated screening procedure for 46 synthetic cannabinoids in serum employing liquid chromatography-quadrupole ion trap mass spectrometry with high-temperature electrospray ionization.

PubMed

Huppertz, Laura M; Kneisel, Stefan; Auwärter, Volker; Kempf, Jürgen

2014-02-01

Considering the vast variety of synthetic cannabinoids and herbal mixtures - commonly known as 'Spice' or 'K2' - on the market and the resulting increase of severe intoxications related to their consumption, there is a need in clinical and forensic toxicology for comprehensive up-to-date screening methods. The focus of this project aimed at developing and implementing an automated screening procedure for the detection of synthetic cannabinoids in serum using a liquid chromatography-ion trap-MS (LC-MS(n)) system and a spectra library-based approach, currently including 46 synthetic cannabinoids and 8 isotope labelled analogues. In the process of method development, a high-temperature ESI source (IonBooster(TM), Bruker Daltonik) and its effects on the ionization efficiency of the investigated synthetic cannabinoids were evaluated and compared to a conventional ESI source. Despite their structural diversity, all investigated synthetic cannabinoids benefitted from high-temperature ionization by showing remarkably higher MS intensities compared to conventional ESI. The employed search algorithm matches retention time, MS and MS(2)/MS(3) spectra. With the utilization of the ionBooster source, limits for the automated detection comparable to cut-off values of routine MRM methods were achieved for the majority of analytes. Even compounds not identified when using a conventional ESI source were detected using the ionBooster-source. LODs in serum range from 0.1 ng/ml to 0.5 ng/ml. The use of parent compounds as analytical targets offers the possibility of instantly adding new emerging compounds to the library and immediately applying the updated method to serum samples, allowing the rapid adaptation of the screening method to ongoing forensic or clinical requirements. The presented approach can also be applied to other specimens, such as oral fluid or hair, and herbal mixtures and was successfully applied to authentic serum samples. Quantitative MRM results of samples with analyte concentrations above the determined LOD were confirmed as positive findings by the presented method. Copyright © 2014 John Wiley & Sons, Ltd.

A novel COLD-PCR/FMCA assay enhances the detection of low-abundance IDH1 mutations in gliomas.

PubMed

Pang, Brendan; Durso, Mary B; Hamilton, Ronald L; Nikiforova, Marina N

2013-03-01

Point mutations in isocitrate dehydrogenase 1 (IDH1) have been identified in many gliomas. The detection of IDH1 mutations becomes challenging on suboptimal glioma biopsies when a limited number of tumor cells is available for analysis. Coamplification at lower denaturing-polymerase chain reaction (COLD-PCR) is a PCR technique that deliberately lowers the denaturing cycle temperature to selectively favor amplification of mutant alleles, allowing for the sensitive detection of low-abundance mutations. We developed a novel COLD-PCR assay on the LightCycler platform (Roche, Applied Science, Indianapolis, IN), using post-PCR fluorescent melting curve analysis (FMCA) for the detection of mutant IDH1 with a detection limit of 1%. Thirty-five WHO grade I to IV gliomas and 9 non-neoplastic brain and spinal cord biopsies were analyzed with this technique and the results were compared with the conventional real-time PCR and the Sanger sequencing analysis. COLD-PCR/FMCA was able to detect the most common IDH1 R132H mutation and rare mutation types including R132H, R132C, R132L, R132S, and R132G mutations. Twenty-five glioma cases were positive for IDH1 mutations by COLD-PCR/FMCA, and 23 gliomas were positive by the conventional real-time PCR and Sanger sequencing. A pilocytic astrocytoma (PA I) and a glioblastoma multiforme (GBM IV) showed low-abundance IDH1 mutations detected by COLD-PCR/FMCA. The remaining 10 glioma and 9 non-neoplastic samples were negative by all the 3 methods. In summary, we report a novel COLD-PCR/FMCA method that provides rapid and sensitive detection of IDH1 mutations in formalin-fixed paraffin-embedded tissue and can be used in the clinical setting to assess the small brain biopsies.

A method based on coffee-ring deposition confocal Raman spectroscopy of analysis of melamine in milk

NASA Astrophysics Data System (ADS)

Tan, Zong; Chen, Da

2016-10-01

In this work, an economical and high-efficiency method for detection of melamine in milk was developed. The enrichment effect of coffee-ring was combined with the micro-region analysis of confocal Raman spectroscopy, in addition, assisted with chemometric algorithmthe. Consequently, a desired result was obtained that the LOD of melamine in this method was 1 ppm, which was excellent because the sensitivity of conventional Raman detection was generally low. Furthermore, the whole process were processed in an easily available condition with almost no chemical reagents consumption, and the chosen substrates for the formation of coffee-ring were reusable. Thus, the method is environmental friendly and has a great potential application in food safety inspection.

Autonomous control systems - Architecture and fundamental issues

NASA Technical Reports Server (NTRS)

Antsaklis, P. J.; Passino, K. M.; Wang, S. J.

1988-01-01

A hierarchical functional autonomous controller architecture is introduced. In particular, the architecture for the control of future space vehicles is described in detail; it is designed to ensure the autonomous operation of the control system and it allows interaction with the pilot and crew/ground station, and the systems on board the autonomous vehicle. The fundamental issues in autonomous control system modeling and analysis are discussed. It is proposed to utilize a hybrid approach to modeling and analysis of autonomous systems. This will incorporate conventional control methods based on differential equations and techniques for the analysis of systems described with a symbolic formalism. In this way, the theory of conventional control can be fully utilized. It is stressed that autonomy is the design requirement and intelligent control methods appear at present, to offer some of the necessary tools to achieve autonomy. A conventional approach may evolve and replace some or all of the `intelligent' functions. It is shown that in addition to conventional controllers, the autonomous control system incorporates planning, learning, and FDI (fault detection and identification).

The application of intraoperative transit time flow measurement to accurately assess anastomotic quality in sequential vein grafting

PubMed Central

Yu, Yang; Zhang, Fan; Gao, Ming-Xin; Li, Hai-Tao; Li, Jing-Xing; Song, Wei; Huang, Xin-Sheng; Gu, Cheng-Xiong

2013-01-01

OBJECTIVES Intraoperative transit time flow measurement (TTFM) is widely used to assess anastomotic quality in coronary artery bypass grafting (CABG). However, in sequential vein grafting, the flow characteristics collected by the conventional TTFM method are usually associated with total graft flow and might not accurately indicate the quality of every distal anastomosis in a sequential graft. The purpose of our study was to examine a new TTFM method that could assess the quality of each distal anastomosis in a sequential graft more reliably than the conventional TTFM approach. METHODS Two TTFM methods were tested in 84 patients who underwent sequential saphenous off-pump CABG in Beijing An Zhen Hospital between April and August 2012. In the conventional TTFM method, normal blood flow in the sequential graft was maintained during the measurement, and the flow probe was placed a few centimetres above the anastomosis to be evaluated. In the new method, blood flow in the sequential graft was temporarily reduced during the measurement by placing an atraumatic bulldog clamp at the graft a few centimetres distal to the anastomosis to be evaluated, while the position of the flow probe remained the same as in the conventional method. This new TTFM method was named the flow reduction TTFM. Graft flow parameters measured by both methods were compared. RESULTS Compared with the conventional TTFM, the flow reduction TTFM resulted in significantly lower mean graft blood flow (P < 0.05); in contrast, yielded significantly higher pulsatility index (P < 0.05). Diastolic filling was not significantly different between the two methods and was >50% in both cases. Interestingly, the flow reduction TTFM identified two defective middle distal anastomoses that the conventional TTFM failed to detect. Graft flows near the defective distal anastomoses were improved substantially after revision. CONCLUSIONS In this study, we found that temporary reduction of graft flow during TTFM seemed to enhance the sensitivity of TTFM to less-than-critical anastomotic defects in a sequential graft and to improve the overall accuracy of the intraoperative assessment of anastomotic quality in sequential vein grafting. PMID:24000314

Detection of influenza antigenic variants directly from clinical samples using polyclonal antibody based proximity ligation assays

PubMed Central

Martin, Brigitte E.; Jia, Kun; Sun, Hailiang; Ye, Jianqiang; Hall, Crystal; Ware, Daphne; Wan, Xiu-Feng

2016-01-01

Identification of antigenic variants is the key to a successful influenza vaccination program. The empirical serological methods to determine influenza antigenic properties require viral propagation. Here a novel quantitative PCR-based antigenic characterization method using polyclonal antibody and proximity ligation assays, or so-called polyPLA, was developed and validated. This method can detect a viral titer that is less than 1000 TCID50/mL. Not only can this method differentiate between different HA subtypes of influenza viruses but also effectively identify antigenic drift events within the same HA subtype of influenza viruses. Applications in H3N2 seasonal influenza data showed that the results from this novel method are consistent with those from the conventional serological assays. This method is not limited to the detection of antigenic variants in influenza but also other pathogens. It has the potential to be applied through a large-scale platform in disease surveillance requiring minimal biosafety and directly using clinical samples. PMID:25546251

Accuracy of contrast-enhanced ultrasound in the detection of bladder cancer

PubMed Central

Nicolau, C; Bunesch, L; Peri, L; Salvador, R; Corral, J M; Mallofre, C; Sebastia, C

2011-01-01

Objective To assess the accuracy contrast-enhanced ultrasound (CEUS) in bladder cancer detection using transurethral biopsy in conventional cystoscopy as the reference standard and to determine whether CEUS improves the bladder cancer detection rate of baseline ultrasound. Methods 43 patients with suspected bladder cancer underwent conventional cystoscopy with transurethral biopsy of the suspicious lesions. 64 bladder cancers were confirmed in 33 out of 43 patients. Baseline ultrasound and CEUS were performed the day before surgery and the accuracy of both techniques for bladder cancer detection and number of detected tumours were analysed and compared with the final diagnosis. Results CEUS was significantly more accurate than ultrasound in determining presence or absence of bladder cancer: 88.37% vs 72.09%. Seven of eight uncertain baseline ultrasound results were correctly diagnosed using CEUS. CEUS sensitivity was also better than that of baseline ultrasound per number of tumours: 65.62% vs 60.93%. CEUS sensitivity for bladder cancer detection was very high for tumours larger than 5 mm (94.7%) but very low for tumours <5 mm (20%) and also had a very low negative predictive value (28.57%) in tumours <5 mm. Conclusion CEUS provided higher accuracy than baseline ultrasound for bladder cancer detection, being especially useful in non-conclusive baseline ultrasound studies. PMID:21123306

Remote NMR/MRI detection of laser polarized gases

DOEpatents

Pines, Alexander; Saxena, Sunil; Moule, Adam; Spence, Megan; Seeley, Juliette A.; Pierce, Kimberly L.; Han, Song-I; Granwehr, Josef

2006-06-13

An apparatus and method for remote NMR/MRI spectroscopy having an encoding coil with a sample chamber, a supply of signal carriers, preferably hyperpolarized xenon and a detector allowing the spatial and temporal separation of signal preparation and signal detection steps. This separation allows the physical conditions and methods of the encoding and detection steps to be optimized independently. The encoding of the carrier molecules may take place in a high or a low magnetic field and conventional NMR pulse sequences can be split between encoding and detection steps. In one embodiment, the detector is a high magnetic field NMR apparatus. In another embodiment, the detector is a superconducting quantum interference device. A further embodiment uses optical detection of Rb--Xe spin exchange. Another embodiment uses an optical magnetometer using non-linear Faraday rotation. Concentration of the signal carriers in the detector can greatly improve the signal to noise ratio.

Depth detection in interactive projection system based on one-shot black-and-white stripe pattern.

PubMed

Zhou, Qian; Qiao, Xiaorui; Ni, Kai; Li, Xinghui; Wang, Xiaohao

2017-03-06

A novel method enabling estimation of not only the screen surface as the conventional one, but the depth information from two-dimensional coordinates in an interactive projection system was proposed in this research. In this method, a one-shot black-and-white stripe pattern from a projector is projected on a screen plane, where the deformed pattern is captured by a charge-coupled device camera. An algorithm based on object/shadow simultaneous detection is proposed for fulfillment of the correspondence. The depth information of the object is then calculated using the triangulation principle. This technology provides a more direct feeling of virtual interaction in three dimensions without using auxiliary equipment or a special screen as interaction proxies. Simulation and experiments are carried out and the results verified the effectiveness of this method in depth detection.

Comparison of computed radiography and conventional radiography in detection of small volume pneumoperitoneum.

PubMed

Marolf, Angela; Blaik, Margaret; Ackerman, Norman; Watson, Elizabeth; Gibson, Nicole; Thompson, Margret

2008-01-01

The role of digital imaging is increasing as these systems are becoming more affordable and accessible. Advantages of computed radiography compared with conventional film/screen combinations include improved contrast resolution and postprocessing capabilities. Computed radiography's spatial resolution is inferior to conventional radiography; however, this limitation is considered clinically insignificant. This study prospectively compared digital imaging and conventional radiography in detecting small volume pneumoperitoneum. Twenty cadaver dogs (15-30 kg) were injected with 0.25, 0.25, and 0.5 ml for 1 ml total of air intra-abdominally, and radiographed sequentially using computed and conventional radiographic technologies. Three radiologists independently evaluated the images, and receiver operating curve (ROC) analysis compared the two imaging modalities. There was no statistical difference between computed and conventional radiography in detecting free abdominal air, but overall computed radiography was relatively more sensitive based on ROC analysis. Computed radiographic images consistently and significantly demonstrated a minimal amount of 0.5 ml of free air based on ROC analysis. However, no minimal air amount was consistently or significantly detected with conventional film. Readers were more likely to detect free air on lateral computed images than the other projections, with no significant increased sensitivity between film/screen projections. Further studies are indicated to determine the differences or lack thereof between various digital imaging systems and conventional film/screen systems.

Paper-based Platform for Urinary Creatinine Detection.

PubMed

Sittiwong, Jarinya; Unob, Fuangfa

2016-01-01

A new paper platform was developed for the colorimetric detection of creatinine. The filter paper was coated with 3-propylsulfonic acid trimethoxysilane and used as the platform. Creatinine in a cationic form was extracted onto the paper via an ion-exchange mechanism and detected through the Jaffé reaction, resulting in a yellow-orange color complex. The color change on the paper could be observed visually, and the quantitative detection of creatinine was achieved through monitoring the color intensity change. The color intensity of creatinine complexes on the paper platform as a function of the creatinine concentration provided a linear range for creatinine detection in the range of 10 - 60 mg L(-1) and a detection limit of 4.2 mg L(-1). The accuracy of the proposed paper-based method was comparable to the conventional standard Jaffé method. This paper platform could be applied for simple and rapid detection of creatinine in human urine samples with a low consumption of reagent.

Full-text automated detection of surgical site infections secondary to neurosurgery in Rennes, France.

PubMed

Campillo-Gimenez, Boris; Garcelon, Nicolas; Jarno, Pascal; Chapplain, Jean Marc; Cuggia, Marc

2013-01-01

The surveillance of Surgical Site Infections (SSI) contributes to the management of risk in French hospitals. Manual identification of infections is costly, time-consuming and limits the promotion of preventive procedures by the dedicated teams. The introduction of alternative methods using automated detection strategies is promising to improve this surveillance. The present study describes an automated detection strategy for SSI in neurosurgery, based on textual analysis of medical reports stored in a clinical data warehouse. The method consists firstly, of enrichment and concept extraction from full-text reports using NOMINDEX, and secondly, text similarity measurement using a vector space model. The text detection was compared to the conventional strategy based on self-declaration and to the automated detection using the diagnosis-related group database. The text-mining approach showed the best detection accuracy, with recall and precision equal to 92% and 40% respectively, and confirmed the interest of reusing full-text medical reports to perform automated detection of SSI.

Trend-Residual Dual Modeling for Detection of Outliers in Low-Cost GPS Trajectories.

PubMed

Chen, Xiaojian; Cui, Tingting; Fu, Jianhong; Peng, Jianwei; Shan, Jie

2016-12-01

Low-cost GPS (receiver) has become a ubiquitous and integral part of our daily life. Despite noticeable advantages such as being cheap, small, light, and easy to use, its limited positioning accuracy devalues and hampers its wide applications for reliable mapping and analysis. Two conventional techniques to remove outliers in a GPS trajectory are thresholding and Kalman-based methods, which are difficult in selecting appropriate thresholds and modeling the trajectories. Moreover, they are insensitive to medium and small outliers, especially for low-sample-rate trajectories. This paper proposes a model-based GPS trajectory cleaner. Rather than examining speed and acceleration or assuming a pre-determined trajectory model, we first use cubic smooth spline to adaptively model the trend of the trajectory. The residuals, i.e., the differences between the trend and GPS measurements, are then further modeled by time series method. Outliers are detected by scoring the residuals at every GPS trajectory point. Comparing to the conventional procedures, the trend-residual dual modeling approach has the following features: (a) it is able to model trajectories and detect outliers adaptively; (b) only one critical value for outlier scores needs to be set; (c) it is able to robustly detect unapparent outliers; and (d) it is effective in cleaning outliers for GPS trajectories with low sample rates. Tests are carried out on three real-world GPS trajectories datasets. The evaluation demonstrates an average of 9.27 times better performance in outlier detection for GPS trajectories than thresholding and Kalman-based techniques.

An eDNA Assay to Monitor a Globally Invasive Fish Species from Flowing Freshwater.

PubMed

Adrian-Kalchhauser, Irene; Burkhardt-Holm, Patricia

2016-01-01

Ponto-Caspian gobies are a flock of five invasive fish species that have colonized freshwaters and brackish waters in Europe and North America. One of them, the round goby Neogobius melanostomus, figures among the 100 worst invaders in Europe. Current methods to detect the presence of Ponto-Caspian gobies involve catching or sighting the fish. These approaches are labor intense and not very sensitive. Consequently, populations are usually detected only when they have reached high densities and when management or containment efforts are futile. To improve monitoring, we developed an assay based on the detection of DNA traces (environmental DNA, or eDNA) of Ponto-Caspian gobies in river water. The assay specifically detects invasive goby DNA and does not react to any native fish species. We apply the assay to environmental samples and demonstrate that parameters such as sampling depth, sampling location, extraction protocol, PCR protocol and PCR inhibition greatly impact detection. We further successfully outline the invasion front of Ponto-Caspian gobies in a large river, the High Rhine in Switzerland, and thus demonstrate the applicability of the assay to lotic environments. The eDNA assay requires less time, equipment, manpower, skills, and financial resources than the conventional monitoring methods such as electrofishing, angling or diving. Samples can be taken by untrained individuals, and the assay can be performed by any molecular biologist on a conventional PCR machine. Therefore, this assay enables environment managers to map invaded areas independently of fishermen's' reports and fish community monitorings.

Ultrasonic sensor based defect detection and characterisation of ceramics.

PubMed

Kesharaju, Manasa; Nagarajah, Romesh; Zhang, Tonzhua; Crouch, Ian

2014-01-01

Ceramic tiles, used in body armour systems, are currently inspected visually offline using an X-ray technique that is both time consuming and very expensive. The aim of this research is to develop a methodology to detect, locate and classify various manufacturing defects in Reaction Sintered Silicon Carbide (RSSC) ceramic tiles, using an ultrasonic sensing technique. Defects such as free silicon, un-sintered silicon carbide material and conventional porosity are often difficult to detect using conventional X-radiography. An alternative inspection system was developed to detect defects in ceramic components using an Artificial Neural Network (ANN) based signal processing technique. The inspection methodology proposed focuses on pre-processing of signals, de-noising, wavelet decomposition, feature extraction and post-processing of the signals for classification purposes. This research contributes to developing an on-line inspection system that would be far more cost effective than present methods and, moreover, assist manufacturers in checking the location of high density areas, defects and enable real time quality control, including the implementation of accept/reject criteria. Copyright © 2013 Elsevier B.V. All rights reserved.

A High Performance Impedance-based Platform for Evaporation Rate Detection.

PubMed

Chou, Wei-Lung; Lee, Pee-Yew; Chen, Cheng-You; Lin, Yu-Hsin; Lin, Yung-Sheng

2016-10-17

This paper describes the method of a novel impedance-based platform for the detection of the evaporation rate. The model compound hyaluronic acid was employed here for demonstration purposes. Multiple evaporation tests on the model compound as a humectant with various concentrations in solutions were conducted for comparison purposes. A conventional weight loss approach is known as the most straightforward, but time-consuming, measurement technique for evaporation rate detection. Yet, a clear disadvantage is that a large volume of sample is required and multiple sample tests cannot be conducted at the same time. For the first time in literature, an electrical impedance sensing chip is successfully applied to a real-time evaporation investigation in a time sharing, continuous and automatic manner. Moreover, as little as 0.5 ml of test samples is required in this impedance-based apparatus, and a large impedance variation is demonstrated among various dilute solutions. The proposed high-sensitivity and fast-response impedance sensing system is found to outperform a conventional weight loss approach in terms of evaporation rate detection.

A case study of application of guided waves for detecting corrosion in pipelines

NASA Astrophysics Data System (ADS)

Rostami, Javad; Safizadeh, Mir Saeed

2012-05-01

Every year noticeable amount of money is spent on fixing and replacing the damaged pipes which carry gas and fuel. Since there is a possibility for a catastrophic failure, knowing the proper time of this repair is of great importance. Because significant proportion of failures is due to wall thinning of pipes because of the corrosion, detecting the wall thinning has been a main part of nondestructive testing of pipes. There are wide variety of NDT techniques to detect this kind of defect such as conventional ultrasonic, eddy current, radiography etc. but some of these techniques, for example conventional ultrasonic needs the insulation of pipes removed and in some other cases such as radiography the test is not done at a reasonable speed. A new method of nondestructive testing of pipes which has the potential to test a long distance in a short period of time and does not need the whole insulation removed, has drawn a lot of attention. In this paper, the ability of ultrasonic guided waves for detecting corrosion in gas pipelines is experimentally investigated.

A Review of Conventional PCR Assays for the Detection of Selected Phytopathogens of Wheat.

PubMed

Kuzdraliński, Adam; Kot, Anna; Szczerba, Hubert; Nowak, Michał; Muszyńska, Marta

2017-01-01

Infection of phyllosphere (stems, leaves, husks, and grains) by pathogenic fungi reduces the wheat yield and grain quality. Detection of the main wheat pathogenic fungi provides information about species composition and allows effective and targeted plant treatment. Since conventional procedures for the detection of these organisms are unreliable and time consuming, diagnostic DNA-based methods are required. Nucleic acid amplification technologies are independent of the morphological and biochemical characteristics of fungi. Microorganisms do not need to be cultured. Therefore, a number of PCR-based methodologies have been developed for the identification of key pathogenic fungi, such as Fusarium spp., Puccinia spp., Zymoseptoria tritici, Parastagonospora nodorum, Blumeria graminis f. sp. tritici, and Pyrenophora tritici-repentis. This article reviews frequently used DNA regions for fungus identification and discusses already known PCR assays for detection of the aforementioned wheat pathogens. We demonstrate that PCR-based wheat pathogen identification assays require further research. In particular, the number of diagnostic tests for Fusarium graminearum, Puccinia spp., and P. tritici-repentis are insufficient. © 2017 S. Karger AG, Basel.

Development and use of a real-time polymerase chain reaction assay for the detection of Ophidiomyces ophiodiicola in snakes.

PubMed

Allender, Matthew C; Bunick, David; Dzhaman, Elena; Burrus, Lucienne; Maddox, Carol

2015-03-01

Fungal pathogens threatening the conservation of wildlife are becoming increasingly common. Since 2008, free-ranging snakes across North America have been experiencing a marked increase in the prevalence of snake fungal disease associated with Ophidiomyces ophiodiicola. Diagnosis has historically relied on histology, microbiology, and conventional polymerase chain reaction (PCR). More sensitive methods are needed to adequately characterize the epidemiology. The current study describes the development of a real-time PCR (qPCR) assay for detecting a segment of the internal transcribed spacer 1 region between the 18S and 5.8S ribosomal RNA gene. The assay was able to detect as few as 1.05 × 10(1) gene copies per reaction. An additional 4 positive cases were detected when comparing a conventional PCR (n = 3) and the qPCR (n = 7) when used on swab samples from 47 eastern massasauga rattlesnakes. The newly developed assay is a sensitive and specific tool for surveillance and monitoring in the conservation of free-ranging snakes. © 2015 The Author(s).

Application of EMA-qPCR as a complementary tool for the detection and monitoring of Legionella in different water systems.

PubMed

Qin, Tian; Tian, Zhengan; Ren, Hongyu; Hu, Guangchun; Zhou, Haijian; Lu, Jinxing; Luo, Chengwang; Liu, Zunyu; Shao, Zhujun

2012-05-01

Legionella are prevalent in human-made water systems and cause legionellosis in humans. Conventional culturing and polymerase chain reaction (PCR) techniques are not sufficiently accurate for the quantitative analysis of live Legionella bacteria in water samples because of the presence of viable but nonculturable cells and dead cells. Here, we report a rapid detection method for viable Legionella that combines ethidium monoazide (EMA) with quantitative real-time PCR (qPCR) and apply this method to detect Legionella in a large number of water samples from different sources. Results yielded that samples treated with 5 μg/ml EMA for 10 min and subsequently exposed to light irradiation for 5 min were optimal for detecting Legionella. EMA treatment before qPCR could block the signal from approximately 4 log(10) of dead cells. When investigating environmental water samples, the percent-positive rate obtained by EMA-qPCR was significantly higher than conventional PCR and culture methods, and slightly lower than qPCR. The bacterial count of Legionella determined by EMA-qPCR were mostly greater than those determined by culture assays and lower than those determined by qPCR. Acceptable correlations were found between the EMA-qPCR and qPCR results for cooling towers, piped water and hot spring water samples (r = 0.849, P < 0.001) and also found between the EMA-qPCR and culture results for hot spring water samples (r = 0.698, P < 0.001). The results indicate that EMA-qPCR could be used as a complementary tool for the detection and monitoring of Legionella in water systems, especially in hot spring water samples.

Direct antigen detection from immunoprecipitated beads using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; a new method for immunobeads-mass spectrometry (iMS).

PubMed

Shimada, Takashi; Toyama, Atsuhiko; Aoki, Chikage; Aoki, Yutaka; Tanaka, Koichi; Sato, Taka-Aki

2011-12-15

One-step detection of biological molecules is one of the principal techniques for clinical diagnosis, and the potential of mass spectrometry for biomarker detection has been a promising new approach in the field of medical sciences. We demonstrate here a new and high-sensitivity method that we termed immunobeads-mass spectrometry (iMS), which combines conventional immunoprecipitation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The key feature of iMS is the MS-compatible condition of immunoprecipitation using detergents with a monosaccaride-C8 alkyl chain or a disaccharide-C10 alkyl chain, and the minimized number of steps required for high-sensitivity detection of target peptides in serum or biological fluid. This was achieved by optimizing the wash buffer and subjecting the immunobeads directly to MALDI-TOF MS analysis. Using this method, we showed that 1 fmol of amyloid beta peptide spiked in serum was readily detectable, demonstrating the powerful tool of iMS as a biomarker detection method. Copyright © 2011 John Wiley & Sons, Ltd.

Novel Hyperspectral Anomaly Detection Methods Based on Unsupervised Nearest Regularized Subspace

NASA Astrophysics Data System (ADS)

Hou, Z.; Chen, Y.; Tan, K.; Du, P.

2018-04-01

Anomaly detection has been of great interest in hyperspectral imagery analysis. Most conventional anomaly detectors merely take advantage of spectral and spatial information within neighboring pixels. In this paper, two methods of Unsupervised Nearest Regularized Subspace-based with Outlier Removal Anomaly Detector (UNRSORAD) and Local Summation UNRSORAD (LSUNRSORAD) are proposed, which are based on the concept that each pixel in background can be approximately represented by its spatial neighborhoods, while anomalies cannot. Using a dual window, an approximation of each testing pixel is a representation of surrounding data via a linear combination. The existence of outliers in the dual window will affect detection accuracy. Proposed detectors remove outlier pixels that are significantly different from majority of pixels. In order to make full use of various local spatial distributions information with the neighboring pixels of the pixels under test, we take the local summation dual-window sliding strategy. The residual image is constituted by subtracting the predicted background from the original hyperspectral imagery, and anomalies can be detected in the residual image. Experimental results show that the proposed methods have greatly improved the detection accuracy compared with other traditional detection method.

QTLs detected for individual sugars and soluble solids content in apple

USDA-ARS?s Scientific Manuscript database

Sweetness is one of the most important fruit quality traits in breeding programs, determining the overall quality and flavor-perception of apples. Selecting for this trait using conventional breeding methods is challenging due to the complexity of its genetic control. In order to improve the efficie...

Accuracy of Conventional and Digital Radiography in Detecting External Root Resorption

PubMed Central

Mesgarani, Abbas; Haghanifar, Sina; Ehsani, Maryam; Yaghub, Samereh Dokhte; Bijani, Ali

2014-01-01

Introduction: External root resorption (ERR) is associated with physiological and pathological dissolution of mineralized tissues by clastic cells and radiography is one of the most important methods in its diagnosis. The aim of this experimental study was to evaluate the accuracy of conventional intraoral radiography (CR) in comparison with digital radiographic techniques, i.e. charge-coupled device (CCD) and photo-stimulable phosphor (PSP) sensors, in detection of ERR. Methods and Materials: This study was performed on 80 extracted human mandibular premolars. After taking separate initial periapical radiographs with CR technique, CCD and PSP sensors, the artificial defects resembling ERR with variable sizes were created in apical half of the mesial, distal and buccal surfaces of the teeth. Ten teeth were used as control samples without any resorption. The radiographs were then repeated with 2 different exposure times and the images were observed by 3 observers. Data were analyzed using SPSS version 17 and chi-squared and Cohen’s Kappa tests with 95% confidence interval (CI=95%). Result: The CCD had the highest percentage of correct assessment compared to the CR and PSP sensors, although the difference was not significant (P=0.39). It was shown that the higher dosage of radiation increases the accuracy of diagnosis; however, it was only significant for CCD sensor (P=0.02). Also, the accuracy of diagnosis increased with the increase in the size of lesion (P=0.001). Conclusion: Statistically significant difference was not observed for accurate detection of ERR by conventional and digital radiographic techniques. PMID:25386202

A Unified Fisher's Ratio Learning Method for Spatial Filter Optimization.

PubMed

Li, Xinyang; Guan, Cuntai; Zhang, Haihong; Ang, Kai Keng

To detect the mental task of interest, spatial filtering has been widely used to enhance the spatial resolution of electroencephalography (EEG). However, the effectiveness of spatial filtering is undermined due to the significant nonstationarity of EEG. Based on regularization, most of the conventional stationary spatial filter design methods address the nonstationarity at the cost of the interclass discrimination. Moreover, spatial filter optimization is inconsistent with feature extraction when EEG covariance matrices could not be jointly diagonalized due to the regularization. In this paper, we propose a novel framework for a spatial filter design. With Fisher's ratio in feature space directly used as the objective function, the spatial filter optimization is unified with feature extraction. Given its ratio form, the selection of the regularization parameter could be avoided. We evaluate the proposed method on a binary motor imagery data set of 16 subjects, who performed the calibration and test sessions on different days. The experimental results show that the proposed method yields improvement in classification performance for both single broadband and filter bank settings compared with conventional nonunified methods. We also provide a systematic attempt to compare different objective functions in modeling data nonstationarity with simulation studies.To detect the mental task of interest, spatial filtering has been widely used to enhance the spatial resolution of electroencephalography (EEG). However, the effectiveness of spatial filtering is undermined due to the significant nonstationarity of EEG. Based on regularization, most of the conventional stationary spatial filter design methods address the nonstationarity at the cost of the interclass discrimination. Moreover, spatial filter optimization is inconsistent with feature extraction when EEG covariance matrices could not be jointly diagonalized due to the regularization. In this paper, we propose a novel framework for a spatial filter design. With Fisher's ratio in feature space directly used as the objective function, the spatial filter optimization is unified with feature extraction. Given its ratio form, the selection of the regularization parameter could be avoided. We evaluate the proposed method on a binary motor imagery data set of 16 subjects, who performed the calibration and test sessions on different days. The experimental results show that the proposed method yields improvement in classification performance for both single broadband and filter bank settings compared with conventional nonunified methods. We also provide a systematic attempt to compare different objective functions in modeling data nonstationarity with simulation studies.

Antimicrobial activity of Eucalyptus camaldulensis essential oils and their interactions with conventional antimicrobial agents against multi-drug resistant Acinetobacter baumannii.

PubMed

Knezevic, Petar; Aleksic, Verica; Simin, Natasa; Svircev, Emilija; Petrovic, Aleksandra; Mimica-Dukic, Neda

2016-02-03

Traditional herbal medicine has become an important issue on the global scale during the past decade. Among drugs of natural origin, special place belongs to essential oils, known as strong antimicrobial agents that can be used to combat antibiotic-resistant bacteria. Eucalyptus camaldulensis leaves are traditional herbal remedy used for various purposes, including treatment of infections. The aim of this study was to determine antimicrobial potential of two E. camaldulensis essential oils against multi-drug resistant (MDR) Acinetobacter baumannii wound isolates and to examine possible interactions of essential oils with conventional antimicrobial agents. Chemical composition of essential oils was determined by gas chromatography-mass spectrometry analysis (GC-MS). MIC values of essential oils against A. baumannii strains were estimated by modified broth microdilution method. The components responsible for antimicrobial activity were detected by bioautographic analysis. The potential synergy between the essential oils and antibiotics (ciprofloxacin, gentamicin and polymyxin B) was examined by checkerboard method and time kill curve. The dominant components of both essential oils were spatulenol, cryptone, p-cimene, 1,8-cineole, terpinen-4-ol and β-pinene. The detected MICs for the E. camaldulensis essential oils were in range from 0.5 to 2 μl mL(-1). The bioautographic assay confirmed antibacterial activity of polar terpene compounds. In combination with conventional antibiotics (ciprofloxacin, gentamicin and polymyxin B), the examined essential oils showed synergistic antibacterial effect in most of the cases, while in some even re-sensitized MDR A. baumannii strains. The synergistic interaction was confirmed by time-kill curves for E. camaldulensis essential oil and polymyxin B combination which reduced bacterial count under detection limit very fast, i.e. after 6h of incubation. The detected anti-A. baumannii activity of E. camaldulensis essential oils justifies traditional use of this plant. The proven E. camaldulensis essential oil synergistic interactions with conventional antibiotics could lead to the development of new treatment strategies of infections caused by MDR A. baumannii strains in the term of antibiotic dose reduction. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Absolute quantification of DNA methylation using microfluidic chip-based digital PCR.

PubMed

Wu, Zhenhua; Bai, Yanan; Cheng, Zule; Liu, Fangming; Wang, Ping; Yang, Dawei; Li, Gang; Jin, Qinghui; Mao, Hongju; Zhao, Jianlong

2017-10-15

Hypermethylation of CpG islands in the promoter region of many tumor suppressor genes downregulates their expression and in a result promotes tumorigenesis. Therefore, detection of DNA methylation status is a convenient diagnostic tool for cancer detection. Here, we reported a novel method for the integrative detection of methylation by the microfluidic chip-based digital PCR. This method relies on methylation-sensitive restriction enzyme HpaII, which cleaves the unmethylated DNA strands while keeping the methylated ones intact. After HpaII treatment, the DNA methylation level is determined quantitatively by the microfluidic chip-based digital PCR with the lower limit of detection equal to 0.52%. To validate the applicability of this method, promoter methylation of two tumor suppressor genes (PCDHGB6 and HOXA9) was tested in 10 samples of early stage lung adenocarcinoma and their adjacent non-tumorous tissues. The consistency was observed in the analysis of these samples using our method and a conventional bisulfite pyrosequencing. Combining high sensitivity and low cost, the microfluidic chip-based digital PCR method might provide a promising alternative for the detection of DNA methylation and early diagnosis of epigenetics-related diseases. Copyright © 2017 Elsevier B.V. All rights reserved.

JPRS Report, Science & Technology, Japan: Symposium on Applications of Advanced Technology: Sensors

DTIC Science & Technology

1992-03-02

attempted as a part of a movement proposed by the [Text] author and his coworkers in 1983. In principle, it is a method that forcibly electrolyzes the...value of the current flowing at that time. The detection and Technology method is similar to that of the conventional phosphorus pentoxide sensor, and...electrolytes 3 ’:.i/ from the standpoint of developing energy-related tech- • ........ !ii. 4 nology. This method of water electrolysis is

Multiplex molecular testing for management of infectious gastroenteritis in a hospital setting: a comparative diagnostic and clinical utility study.

PubMed

Halligan, E; Edgeworth, J; Bisnauthsing, K; Bible, J; Cliff, P; Aarons, E; Klein, J; Patel, A; Goldenberg, S

2014-08-01

Laboratory diagnosis and clinical management of inpatients with diarrhoea is complex and time consuming. Tests are often requested sequentially and undertaken in different laboratories. This causes prolonged unnecessary presumptive isolation of patients, because most cases are non-infectious. A molecular multiplex test (Luminex(®) Gastrointestinal Pathogen Panel (GPP)) was compared with conventional testing over 8 months to determine diagnostic accuracy, turnaround times, laboratory costs, use of isolation facilities and user acceptability. A total of 262 (12%) patients had a pathogen detected by conventional methods compared with 483 (22.1%) by GPP. Most additional cases were detected in patients developing symptoms in the first 4 days of admission. Additional cases were detected because of presumed improved diagnostic sensitivity but also because clinicians had not requested the correct pathogen. Turnaround time (41.8 h) was faster than bacterial culture (66.5 h) and parasite investigation (66.5 h) but slower than conventional testing for Clostridium difficile (17.3 h) and viruses (27 h). The test could allow simplified requesting by clinicians and a consolidated laboratory workflow, reducing the overall number of specimens received by the laboratory. A total of 154 isolation days were saved at an estimated cost of £30 800. Consumables and labour were estimated at £150 641 compared with £63 431 for conventional testing. Multiplex molecular testing using a panel of targets allowed enhanced detection and a consolidated laboratory workflow. This is likely to be of greater benefit to cases that present within the first 4 days of hospital admission. © 2013 The Authors Clinical Microbiology and Infection © 2013 European Society of Clinical Microbiology and Infectious Diseases.

[Application of array-based comparative genomic hybridization technique in genetic analysis of patients with spontaneous abortion].

PubMed

Chu, Y; Wu, D; Hou, Q F; Huo, X D; Gao, Y; Wang, T; Wang, H D; Yang, Y L; Liao, S X

2016-08-25

To investigate the value of array-based comparative genomic hybridization (array-CGH) technique for the detection of chromosomal analysis of miscarried embryo, and to provide genetic counseling for couples with spontaneous abortion. Totally 382 patients who underwent miscarriage were enrolled in this study. All aborted tissues were analyzed with conventional cytogenetic karyotyping and array-CGH, respectively. Through genetic analysis, all of the 382 specimens were successfully analyzed by array-CGH (100.0%, 382/382), and the detection rate of chromosomal aberrations was 46.6% (178/382). However, conventional karyotype analysis was successfully performed in 281 cases (73.6%, 281/382), and 113 (40.2%, 113/281) were found with chromosomal aberrations. Of these 178 samples identified by array-CGH, 163 samples (91.6%, 163/178) were aneuploidy, 15 samples (8.4%, 15/178) were segmental deletion and (or) duplication cases. Four of 10 cases with small segmental deletion and duplication were validated to be transferred from their fathers or mathers who were carriers of submicroscopic reciprocal translocation. Of these 113 abnormal karyotypes founded by conventional karyotyping, 108 cases (95.6%, 108/113) were aneuploidy and 5 cases (4.4%, 5/113) had chromosome structural aberrations. Most array-CGH results were consistent with conventional karyotyping but with 3 cases of discrepancy, which included 2 cases of triploids, 1 case of low-level mosaicism that undetcted by array-CGH. Compared with conventional karyotyping, there is an increased detection rate of chromosomal abnormalities when array-CGH is used to analyse the products of conception, primarilly because of its sucess with nonviable tissues. It could be a first-line method to determine the reason of miscarrage with higher accuracy and sensitivity.

Rapid diagnostic tests duo as alternative to conventional serological assays for conclusive Chagas disease diagnosis.

PubMed

Egüez, Karina E; Alonso-Padilla, Julio; Terán, Carolina; Chipana, Zenobia; García, Wilson; Torrico, Faustino; Gascon, Joaquim; Lozano-Beltran, Daniel-Franz; Pinazo, María-Jesús

2017-04-01

Chagas disease is caused by the parasite Trypanosoma cruzi. It affects several million people, mainly in Latin America, and severe cardiac and/or digestive complications occur in ~30% of the chronically infected patients. Disease acute stage is mostly asymptomatic and infection goes undiagnosed. In the chronic phase direct parasite detection is hampered due to its concealed presence and diagnosis is achieved by serological methods, like ELISA or indirect hemagglutination assays. Agreement in at least two tests must be obtained due to parasite wide antigenic variability. These techniques require equipped labs and trained personnel and are not available in distant regions. As a result, many infected people often remain undiagnosed until it is too late, as the two available chemotherapies show diminished efficacy in the advanced chronic stage. Easy-to-use rapid diagnostic tests have been developed to be implemented in remote areas as an alternative to conventional tests. They do not need electricity, nor cold chain, they can return results within an hour and some even work with whole blood as sample, like Chagas Stat-Pak (ChemBio Inc.) and Chagas Detect Plus (InBIOS Inc.). Nonetheless, in order to qualify a rapidly diagnosed positive patient for treatment, conventional serological confirmation is obligatory, which might risk its start. In this study two rapid tests based on distinct antigen sets were used in parallel as a way to obtain a fast and conclusive Chagas disease diagnosis using whole blood samples. Chagas Stat-Pak and Chagas Detect Plus were validated by comparison with three conventional tests yielding 100% sensitivity and 99.3% specificity over 342 patients seeking Chagas disease diagnosis in a reference centre in Sucre (Bolivia). Combined used of RDTs in distant regions could substitute laborious conventional serology, allowing immediate treatment and favouring better adhesion to it.

Evaluation of detection methods for screening meat and poultry products for the presence of foodborne pathogens.

PubMed

Bohaychuk, Valerie M; Gensler, Gary E; King, Robin K; Wu, John T; McMullen, Lynn M

2005-12-01

Rapid and molecular technologies such as enzyme-linked immunosorbent assay (ELISA), PCR, and lateral flow immunoprecipitation can reduce the time and labor involved in screening food products for the presence of pathogens. These technologies were compared with conventional culture methodology for the detection of Salmonella, Campylobacter, Listeria, and Escherichia coli O157:H7 inoculated in raw and processed meat and poultry products. Recommended protocols were modified so that the same enrichment broths used in the culture methods were also used in the ELISA, PCR, and lateral flow immunoprecipitation assays. The percent agreement between the rapid technologies and culture methods ranged from 80 to 100% depending on the pathogen detected and the method used. ELISA, PCR, and lateral flow immunoprecipitation all performed well, with no statistical difference, compared with the culture method for the detection of E. coli O157:H7. ELISA performed better for the detection of Salmonella, with sensitivity and specificity rates of 100%. PCR performed better for the detection of Campylobacter jejuni, with 100% agreement to the culture method. PCR was highly sensitive for the detection of all the foodborne pathogens tested except Listeria monocytogenes. Although the lateral flow immunoprecipitation tests were statistically different from the culture methods for Salmonella and Listeria because of false-positive results, the tests did not produce any false negatives, indicating that this method would be suitable for screening meat and poultry products for these pathogens.

Sensitive SERS detection of lead ions via DNAzyme based quadratic signal amplification.

PubMed

Tian, Aihua; Liu, Yu; Gao, Jian

2017-08-15

Highly sensitive detection of Pb 2+ is very necessary for water quality control, clinical toxicology, and industrial monitoring. In this work, a simple and novel DNAzyme-based SERS quadratic amplification method is developed for the detection of Pb 2+ . This strategy possesses some remarkable features compared to the conventional DNAzyme-based SERS methods, which are as follows: (i) Coupled DNAzyme-activated hybridization chain reaction (HCR) with bio barcodes; a quadratic amplification method is designed using the unique catalytic selectivity of DNAzyme. The SERS signal is significantly amplified. This method is rapid with a detection time of 2h. (ii) The problem of high background induced by excess bio barcodes is circumvented by using magnetic beads (MBs) as the carrier of signal-output products, and this sensing system is simple in design and can easily be carried out by simple mixing and incubation. Given the unique and attractive characteristics, a simple and universal strategy is designed to accomplish sensitive detection of Pb 2+ . The detection limit of Pb 2+ via SERS detection is 70 fM, with the linear range from 1.0×10 -13 M to 1.0×10 -7 M. The method can be further extended to the quantitative detection of a variety of targets by replacing the lead-responsive DNAzyme with other functional DNA. Copyright © 2017 Elsevier B.V. All rights reserved.

Current Technical Approaches for the Early Detection of Foodborne Pathogens: Challenges and Opportunities.

PubMed

Cho, Il-Hoon; Ku, Seockmo

2017-09-30

The development of novel and high-tech solutions for rapid, accurate, and non-laborious microbial detection methods is imperative to improve the global food supply. Such solutions have begun to address the need for microbial detection that is faster and more sensitive than existing methodologies (e.g., classic culture enrichment methods). Multiple reviews report the technical functions and structures of conventional microbial detection tools. These tools, used to detect pathogens in food and food homogenates, were designed via qualitative analysis methods. The inherent disadvantage of these analytical methods is the necessity for specimen preparation, which is a time-consuming process. While some literature describes the challenges and opportunities to overcome the technical issues related to food industry legal guidelines, there is a lack of reviews of the current trials to overcome technological limitations related to sample preparation and microbial detection via nano and micro technologies. In this review, we primarily explore current analytical technologies, including metallic and magnetic nanomaterials, optics, electrochemistry, and spectroscopy. These techniques rely on the early detection of pathogens via enhanced analytical sensitivity and specificity. In order to introduce the potential combination and comparative analysis of various advanced methods, we also reference a novel sample preparation protocol that uses microbial concentration and recovery technologies. This technology has the potential to expedite the pre-enrichment step that precedes the detection process.

[Evaluation of PNA-FISH method for direct identification of Candida species in blood culture samples and its potential impact on guidance of antifungal therapy].

PubMed

Doğan, Özlem; İnkaya, Ahmet Çağkan; Gülmez, Dolunay; Uzun, Ömrüm; Akova, Murat; Arıkan Akdağlı, Sevtap

2016-10-01

Early antifungal therapy has a major influence on survival in candidemia. Rapid identification of the species has importance for the treatment, prediction of the species-specific primary resistance and variable antifungal susceptibility. Recently, molecular-based methods attempt to reduce the time between the positive signal of a blood culture and identification of the fungus. PNA-FISH (Peptide nucleic acid fluorescence in situ hybridization) assay distinguishes a number of frequently isolated Candida species in groups following the growth in blood culture. The aim of this study was to investigate the correlation of the species identified by PNA-FISH with conventional identification methods in yeast positive blood cultures and its influence on the selection of antifungal therapy. Specimens of adult patients diagnosed as yeast with Gram stain in signal-positive blood cultures between August to December 2013, were included in the study. The strains were concomitantly cultivated by subculturing from the blood culture bottles onto solid media and identified by conventional methods (germ tube test, ID32C and morphology on cornmeal Tween 80 agar). Rapid species identification was performed by Yeast Traffic Light PNA-FISH, which generates green flourescence for Candida albicans and Candida parapsilosis, yellow for Candida tropicalis, and red for Candida krusei and Candida glabrata. C.tropicalis was identified as a single species whereas the others were identified in pairs. The time points when the yeast positive blood culture bottle was received by the mycology laboratory and reporting of the species identification results by PNA-FISH and the conventional methods were recorded. Seven C.albicans, six C.glabrata, three C.parapsilosis, one C.tropicalis, one C.krusei, one Cryptococcus neoformans, one Saprochaete capitata (Blastoschizomyces capitatus), one C.albicans and Candida dubliniensis, one C.krusei and C.dubliniensis, and one C.glabrata and C.parapsilosis were identified by conventional methods in 23 specimens. Results of PNA-FISH and conventional methods were in full agreement in 19 of the 23 specimens (82.6%). Two specimens were negative by PNA-FISH and yielded S.capitata and C.neoformans which were not included in the test panel. In three specimens that were infected with multiple species, PNA-FISH detected only one of the species. On the other hand and in one specimen, PNA-FISH detected a second species (C.glabrata or C.krusei) that could not be isolated and identified conventionally. Species identification were obtained 72 hours (mean) earlier with PNA-FISH. PNA-FISH provided accurate species identification that were consistent with conventional methods. However and expectedly, it failed to detect species that were not included in the test panel. During the study period, 13 of the 23 patients have passed away. Apart from six patients died prior to blood culture positivity and the one that could not get any antifungal therapy during hospital stay, 16 patients received antifungal treatment. Of sixteen patients who received antifungal therapy, initial antifungal treatment was fluconazole for five and echinocandin for 10 patients. Fluconazole and amphotericin B combination was preferred for one patient. In this study, PNA-FISH result had an influence on the modification of the antifungal treatment of only for one patient in accordance with the clinical findings. We conclude that the utility of PNA-FISH method appeared to be limited in our center since the assay cannot differentiate C.albicans and C.parapsilosis, the two commonly isolated species among our candidemia isolates. However, advantages of the assay might be more pronounced for the centers where C.glabrata is a relatively more frequent species.

Are You Ready for [a] Roundup?--What Chemistry Has to Do with Genetic Modifications

NASA Astrophysics Data System (ADS)

Pöpping, Bert

2001-06-01

Genetically modified crops are grown in most parts of the world nowadays. These transgenic plants have new properties such as herbicide tolerance or insect resistance that often cannot be introduced by conventional breeding. Using examples of very common transgenic varieties, the article explains how the knowledge of metabolic pathways and genetic information is used to design these plants and how the same knowledge is used to detect them. It reviews why detection of genetic modifications in plants has become necessary and describes the most common detection methods, from immunological assays to polymerase chain reaction and real-time detection.

Serine Protease Zymography: Low-Cost, Rapid, and Highly Sensitive RAMA Casein Zymography.

PubMed

Yasumitsu, Hidetaro

2017-01-01

To detect serine protease activity by zymography, casein and CBB stain have been used as a substrate and a detection procedure, respectively. Casein zymography has been using substrate concentration at 1 mg/mL and employing conventional CBB stain. Although ordinary casein zymography provides reproducible results, it has several disadvantages including time-consuming and relative low sensitivity. Improved casein zymography, RAMA casein zymography, is rapid and highly sensitive. RAMA casein zymography completes the detection process within 1 h after incubation and increases the sensitivity at least by tenfold. In addition to serine protease, the method also detects metalloprotease 7 (MMP7, Matrilysin) with high sensitivity.

Optical detection of tracer species in strongly scattering media.

PubMed

Brauser, Eric M; Rose, Peter E; McLennan, John D; Bartl, Michael H

2015-03-01

A combination of optical absorption and scattering is used to detect tracer species in a strongly scattering medium. An optical setup was developed, consisting of a dual-beam scattering detection scheme in which sample scattering beam overlaps with the characteristic absorption feature of quantum dot tracer species, while the reference scattering beam is outside any absorption features of the tracer. This scheme was successfully tested in engineered breakthrough tests typical of wastewater and subsurface fluid analysis, as well as in batch analysis of oil and gas reservoir fluids and biological samples. Tracers were detected even under highly scattering conditions, conditions in which conventional absorption or fluorescence methods failed.

Conventional forceps or hot biopsy: comparative study of two methods in diagnosis of endobronchial lesions.

PubMed

Jabbari, Hamidreza; Fakhri, Mohammad; Lotfaliani, Mojtaba; Kiani, Arda

2013-01-01

It is suggested that hot electrocoagulation-enabled forceps (hot biopsy) may reduce hemorrhage risk after the biopsy in endobronchial tumors. The main concern in this method is possible reduction of the specimen's quality. To compare the procedure related hemorrhage with hot biopsy and conventional forceps biopsy and the diagnostic quality of the obtained specimens with either technique. In this prospective study, assessment of the biopsy samples and quantity of hemorrhage were done in a blind fashion. At first, for each patient a definite clinical diagnosis was made based on pathologic examination of all available samples, clinical data, and imaging findings. Then, second pathologist reviewed all samples to evaluate the quality of the samples. A total of 36 patients with endobronchial lesions were included in this study. Definite diagnosis was made in 83% of the patients. Diagnostic yield of the two methods were not statistically different, while the mean hemorrhage grades of all hot biopsy protocols were significantly lower as compared to that of conventional biopsy (p=0.003, p<0.001 and p<0.001 for 10,20and40 voltages respectively). No significant difference was detected between the qualities of specimens obtained by hot biopsy methods in comparison with conventional biopsy (p>0.05 for all three voltages). Hot biopsy can be a valuable alternative to forceps biopsy in evaluating endobronchial lesions.

Evaluation of phage assay for rapid phenotypic detection of rifampicin resistance in Mycobacterium tuberculosis

PubMed Central

Yzquierdo, Sergio Luis; Lemus, Dihadenys; Echemendia, Miguel; Montoro, Ernesto; McNerney, Ruth; Martin, Anandi; Palomino, Juan Carlos

2006-01-01

Background Conventional methods for susceptibility testing require several months before results can be reported. However, rapid methods to determine drug susceptibility have been developed recently. Phage assay have been reported as a rapid useful tools for antimicrobial susceptibility testing. The aim of this study was to apply the Phage assay for rapid detection of resistance on Mycobacterium tuberculosis strains in Cuba. Methods Phage D29 assay was performed on 102 M. tuberculosis strains to detect rifampicin resistance. The results were compared with the proportion method (gold standard) to evaluate the sensitivity and specificity of Phage assay. Results Phage assay results were available in 2 days whereas Proportion Methods results were obtain in 42 days. A total of 44 strains were detected as rifampicin resistant by both methods. However, one strains deemed resistant by Proportion Methods was susceptible by Phage assay. The sensitivity and specificity of Phage assay were 97.8 % and 100% respectively. Conclusion Phage assay provides rapid and reliable results for susceptibility testing; it's easy to perform, requires no specialized equipment and is applicable to drug susceptibility testing in low income countries where tuberculosis is a major public health problem. PMID:16630356

Measurement technology of RF interference current in high current system

NASA Astrophysics Data System (ADS)

Zhao, Zhihua; Li, Jianxuan; Zhang, Xiangming; Zhang, Lei

2018-06-01

Current probe is a detection method commonly used in electromagnetic compatibility. With the development of power electronics technology, the power level of power conversion devices is constantly increasing, and the power current of the electric energy conversion device in the electromagnetic launch system can reach 10kA. Current probe conventionally used in EMC (electromagnetic compatibility) detection cannot meet the test requirements on high current system due to the magnetic saturation problem. The conventional high current sensor is also not suitable for the RF (Radio Frequency) interference current measurement in high current power device due to the high noise level in the output of active amplifier. In this paper, a passive flexible current probe based on Rogowski coil and matching resistance is proposed that can withstand high current and has low noise level, to solve the measurement problems of interference current in high current power converter. And both differential mode and common mode current detection can be easily carried out with the proposed probe because of the probe's flexible structure.

Application of artificial neural network to search for gravitational-wave signals associated with short gamma-ray bursts

NASA Astrophysics Data System (ADS)

Kim, Kyungmin; Harry, Ian W.; Hodge, Kari A.; Kim, Young-Min; Lee, Chang-Hwan; Lee, Hyun Kyu; Oh, John J.; Oh, Sang Hoon; Son, Edwin J.

2015-12-01

We apply a machine learning algorithm, the artificial neural network, to the search for gravitational-wave signals associated with short gamma-ray bursts (GRBs). The multi-dimensional samples consisting of data corresponding to the statistical and physical quantities from the coherent search pipeline are fed into the artificial neural network to distinguish simulated gravitational-wave signals from background noise artifacts. Our result shows that the data classification efficiency at a fixed false alarm probability (FAP) is improved by the artificial neural network in comparison to the conventional detection statistic. Specifically, the distance at 50% detection probability at a fixed false positive rate is increased about 8%-14% for the considered waveform models. We also evaluate a few seconds of the gravitational-wave data segment using the trained networks and obtain the FAP. We suggest that the artificial neural network can be a complementary method to the conventional detection statistic for identifying gravitational-wave signals related to the short GRBs.

Detection of Listeria monocytogenes from selective enrichment broth using MALDI-TOF Mass Spectrometry.

PubMed

Jadhav, Snehal; Sevior, Danielle; Bhave, Mrinal; Palombo, Enzo A

2014-01-31

Conventional methods used for primary detection of Listeria monocytogenes from foods and subsequent confirmation of presumptive positive samples involve prolonged incubation and biochemical testing which generally require four to five days to obtain a result. In the current study, a simple and rapid proteomics-based MALDI-TOF MS approach was developed to detect L. monocytogenes directly from selective enrichment broths. Milk samples spiked with single species and multiple species cultures were incubated in a selective enrichment broth for 24h, followed by an additional 6h secondary enrichment. As few as 1 colony-forming unit (cfu) of L. monocytogenes per mL of initial selective broth culture could be detected within 30h. On applying the same approach to solid foods previously implicated in listeriosis, namely chicken pâté, cantaloupe and Camembert cheese, detection was achieved within the same time interval at inoculation levels of 10cfu/mL. Unlike the routine application of MALDI-TOF MS for identification of bacteria from solid media, this study proposes a cost-effective and time-saving detection scheme for direct identification of L. monocytogenes from broth cultures.This article is part of a Special Issue entitled: Trends in Microbial Proteomics. Globally, foodborne diseases are major causes of illness and fatalities in humans. Hence, there is a continual need for reliable and rapid means for pathogen detection from food samples. Recent applications of MALDI-TOF MS for diagnostic microbiology focused on detection of microbes from clinical specimens. However, the current study has emphasized its use as a tool for detecting the major foodborne pathogen, Listeria monocytogenes, directly from selective enrichment broths. This proof-of-concept study proposes a detection scheme that is more rapid and simple compared to conventional methods of Listeria detection. Very low levels of the pathogen could be identified from different food samples post-enrichment in selective enrichment broths. Use of this scheme will facilitate rapid and cost-effective testing for this important foodborne pathogen. © 2013.

The role of Fluorine-18-Fluorodeoxyglucose positron emission tomography in staging and restaging of patients with osteosarcoma

PubMed Central

Quartuccio, Natale; Treglia, Giorgio; Salsano, Marco; Mattoli, Maria Vittoria; Muoio, Barbara; Piccardo, Arnoldo; Lopci, Egesta; Cistaro, Angelina

2013-01-01

Background The objective of this study is to systematically review the role of positron emission tomography (PET) and PET/computed tomography (PET/CT) with Fluorine-18-Fluorodeoxyglucose (FDG) in patients with osteosarcoma (OS). Methods A comprehensive literature search of published studies through October 10th, 2012 in PubMed/MEDLINE, Embase and Scopus databases regarding whole-body FDG-PET and FDG-PET/CT in patients with OS was performed. Results We identified 13 studies including 289 patients with OS. With regard to the staging and restaging of OS, the diagnostic performance of FDG-PET and PET/CT seem to be high; FDG-PET and PET/CT seem to be superior to bone scintigraphy and conventional imaging methods in detecting bone metastases; conversely, spiral CT seems to be superior to FDG-PET in detecting pulmonary metastases from OS Conclusions Metabolic imaging may provide additional information in the evaluation of OS patients. The combination of FDG-PET or FDG-PET/CT with conventional imaging methods seems to be a valuable tool in the staging and restaging of OS and may have a relevant impact on the treatment planning. PMID:23801904

Evaluation of maternal serum alpha-foetoprotein assay using dry blood spot samples.

PubMed

González, C; Guerrero, J M; Elorza, F L; Molinero, P; Goberna, R

1988-02-01

The quantification of alpha-foetoprotein in dry blood spots from pregnant women was evaluated, using a conventional radioimmunoassay (RIA) with a monospecific antibody. The stability of alpha-foetoprotein in dry blood spots on filter paper was evaluated with respect to mailing, distances travelled, and the existence of high summer temperatures in our region. The results obtained show that the blood alpha-foetoprotein is stable on dry filter spots sent by mail and is stable for up to four weeks at 4, 25 and 37 degrees C. The analytical method used has a minimal detectable concentration of 10 +/- 1.9 international kilo-units/l. Both inter- and intra-assay variabilities are smaller than 10% and this method can provide results comparable with those of conventional serum assays. Results from dry blood spots and serum samples (the latter analysed by both RIA and two-site enzyme immunoassay) exhibited a good correlation (r = 0.98 and r = 0.97, p less than 0.001). The design of the assay and the nature of the samples make this method suitable for a screening programmes for the antenatal detection of open neural tube defects.

Lead-lag cross-sectional structure and detection of correlated anticorrelated regime shifts: Application to the volatilities of inflation and economic growth rates

NASA Astrophysics Data System (ADS)

Zhou, Wei-Xing; Sornette, Didier

2007-07-01

We have recently introduced the “thermal optimal path” (TOP) method to investigate the real-time lead-lag structure between two time series. The TOP method consists in searching for a robust noise-averaged optimal path of the distance matrix along which the two time series have the greatest similarity. Here, we generalize the TOP method by introducing a more general definition of distance which takes into account possible regime shifts between positive and negative correlations. This generalization to track possible changes of correlation signs is able to identify possible transitions from one convention (or consensus) to another. Numerical simulations on synthetic time series verify that the new TOP method performs as expected even in the presence of substantial noise. We then apply it to investigate changes of convention in the dependence structure between the historical volatilities of the USA inflation rate and economic growth rate. Several measures show that the new TOP method significantly outperforms standard cross-correlation methods.

Improvement in QEPAS system utilizing a second harmonic based wavelength calibration technique

NASA Astrophysics Data System (ADS)

Zhang, Qinduan; Chang, Jun; Wang, Fupeng; Wang, Zongliang; Xie, Yulei; Gong, Weihua

2018-05-01

A simple laser wavelength calibration technique, based on second harmonic signal, is demonstrated in this paper to improve the performance of quartz enhanced photoacoustic spectroscopy (QEPAS) gas sensing system, e.g. improving the signal to noise ratio (SNR), detection limit and long-term stability. Constant current, corresponding to the gas absorption line, combining f/2 frequency sinusoidal signal are used to drive the laser (constant driving mode), a software based real-time wavelength calibration technique is developed to eliminate the wavelength drift due to ambient fluctuations. Compared to conventional wavelength modulation spectroscopy (WMS), this method allows lower filtering bandwidth and averaging algorithm applied to QEPAS system, improving SNR and detection limit. In addition, the real-time wavelength calibration technique guarantees the laser output is modulated steadily at gas absorption line. Water vapor is chosen as an objective gas to evaluate its performance compared to constant driving mode and conventional WMS system. The water vapor sensor was designed insensitive to the incoherent external acoustic noise by the numerical averaging technique. As a result, the SNR increases 12.87 times in wavelength calibration technique based system compared to conventional WMS system. The new system achieved a better linear response (R2 = 0 . 9995) in concentration range from 300 to 2000 ppmv, and achieved a minimum detection limit (MDL) of 630 ppbv.

Detection of Gastrointestinal Pathogens from Stool Samples on Hemoccult Cards by Multiplex PCR

PubMed Central

Schlenker, Nicklas; Bauer, Malkin; Helfrich, Kerstin; Mengele, Carolin; Löscher, Thomas; Nothdurft, Hans Dieter; Bretzel, Gisela; Beissner, Marcus

2017-01-01

Purpose. Up to 30% of international travelers are affected by travelers' diarrhea (TD). Reliable data on the etiology of TD is lacking. Sufficient laboratory capacity at travel destinations is often unavailable and transporting conventional stool samples to the home country is inconvenient. We evaluated the use of Hemoccult cards for stool sampling combined with a multiplex PCR for the detection of model viral, bacterial, and protozoal TD pathogens. Methods. Following the creation of serial dilutions for each model pathogen, last positive dilution steps (LPDs) and thereof calculated last positive sample concentrations (LPCs) were compared between conventional stool samples and card samples. Furthermore, card samples were tested after a prolonged time interval simulating storage during a travel duration of up to 6 weeks. Results. The LPDs/LPCs were comparable to testing of conventional stool samples. After storage on Hemoccult cards, the recovery rate was 97.6% for C. jejuni, 100% for E. histolytica, 97.6% for norovirus GI, and 100% for GII. Detection of expected pathogens was possible at weekly intervals up to 42 days. Conclusion. Stool samples on Hemoccult cards stored at room temperature can be used in combination with a multiplex PCR as a reliable tool for testing of TD pathogens. PMID:28408937

Laparoscopic Single Site Adrenalectomy Using a Conventional Laparoscope and Instrumentation

PubMed Central

Colon, Modesto J; LeMasters, Patrick; Newell, Phillipa; Divino, Celia; Weber, Kaare J.

2011-01-01

Background and Objectives: We present a case of Laparoendoscopic Single Site Surgery (LESS) left adrenalectomy performed with a conventional laparoscope and instruments. Methods: A 45-year-old male was diagnosed with hyperaldosteronism. Computed tomography detected a left adrenal nodule. Bilateral adrenal vein sampling was consistent with a left-sided source for hyperaldosteronism. Results: Total operative time for LESS left adrenalectomy was 120 minutes. The surgery was performed with conventional instruments, a standard 5-mm laparoscope, and a SILS port, with no additional incisions or trocars needed. No complications occurred, and the patient reported an uneventful recovery. Conclusions: LESS adrenalectomy is a feasible procedure. Although articulating instruments and laparoscopes may offer advantages, LESS adrenalectomy can be done without these. PMID:21902983

Detecting and quantifying stellar magnetic fields. Sparse Stokes profile approximation using orthogonal matching pursuit

NASA Astrophysics Data System (ADS)

Carroll, T. A.; Strassmeier, K. G.

2014-03-01

Context. In recent years, we have seen a rapidly growing number of stellar magnetic field detections for various types of stars. Many of these magnetic fields are estimated from spectropolarimetric observations (Stokes V) by using the so-called center-of-gravity (COG) method. Unfortunately, the accuracy of this method rapidly deteriorates with increasing noise and thus calls for a more robust procedure that combines signal detection and field estimation. Aims: We introduce an estimation method that provides not only the effective or mean longitudinal magnetic field from an observed Stokes V profile but also uses the net absolute polarization of the profile to obtain an estimate of the apparent (i.e., velocity resolved) absolute longitudinal magnetic field. Methods: By combining the COG method with an orthogonal-matching-pursuit (OMP) approach, we were able to decompose observed Stokes profiles with an overcomplete dictionary of wavelet-basis functions to reliably reconstruct the observed Stokes profiles in the presence of noise. The elementary wave functions of the sparse reconstruction process were utilized to estimate the effective longitudinal magnetic field and the apparent absolute longitudinal magnetic field. A multiresolution analysis complements the OMP algorithm to provide a robust detection and estimation method. Results: An extensive Monte-Carlo simulation confirms the reliability and accuracy of the magnetic OMP approach where a mean error of under 2% is found. Its full potential is obtained for heavily noise-corrupted Stokes profiles with signal-to-noise variance ratios down to unity. In this case a conventional COG method yields a mean error for the effective longitudinal magnetic field of up to 50%, whereas the OMP method gives a maximum error of 18%. It is, moreover, shown that even in the case of very small residual noise on a level between 10-3 and 10-5, a regime reached by current multiline reconstruction techniques, the conventional COG method incorrectly interprets a large portion of the residual noise as a magnetic field, with values of up to 100 G. The magnetic OMP method, on the other hand, remains largely unaffected by the noise, regardless of the noise level the maximum error is no greater than 0.7 G.

A comparison study on microwave-assisted extraction of Artemisia sphaerocephala polysaccharides with conventional method: Molecule structure and antioxidant activities evaluation.

PubMed

Wang, Junlong; Zhang, Ji; Wang, Xiaofang; Zhao, Baotang; Wu, Yiqian; Yao, Jian

2009-12-01

The conventional extraction methods for polysaccharides were time-consuming, laborious and energy-consuming. Microwave-assisted extraction (MAE) technique was employed for the extraction of Artemisia sphaerocephala polysaccharides (ASP), which is a traditional Chinese food. The extracting parameters were optimized by Box-Behnken design. In microwave heating process, a decrease in molecular weight (M(w)) was detected in SEC-LLS measurement. A d(f) value of 2.85 indicated ASP using MAE exhibited as a sphere conformation of branched clusters in aqueous solution. Furthermore, it showed stronger antioxidant activities compared with hot water extraction. The data obtained showed that the molecular weights played a more important role in antioxidant activities.

a Single-Exposure Dual-Energy Computed Radiography Technique for Improved Nodule Detection and Classification in Chest Imaging

NASA Astrophysics Data System (ADS)

Zink, Frank Edward

The detection and classification of pulmonary nodules is of great interest in chest radiography. Nodules are often indicative of primary cancer, and their detection is particularly important in asymptomatic patients. The ability to classify nodules as calcified or non-calcified is important because calcification is a positive indicator that the nodule is benign. Dual-energy methods offer the potential to improve both the detection and classification of nodules by allowing the formation of material-selective images. Tissue-selective images can improve detection by virtue of the elimination of obscuring rib structure. Bone -selective images are essentially calcium images, allowing classification of the nodule. A dual-energy technique is introduced which uses a computed radiography system to acquire dual-energy chest radiographs in a single-exposure. All aspects of the dual-energy technique are described, with particular emphasis on scatter-correction, beam-hardening correction, and noise-reduction algorithms. The adaptive noise-reduction algorithm employed improves material-selective signal-to-noise ratio by up to a factor of seven with minimal sacrifice in selectivity. A clinical comparison study is described, undertaken to compare the dual-energy technique to conventional chest radiography for the tasks of nodule detection and classification. Observer performance data were collected using the Free Response Observer Characteristic (FROC) method and the bi-normal Alternative FROC (AFROC) performance model. Results of the comparison study, analyzed using two common multiple observer statistical models, showed that the dual-energy technique was superior to conventional chest radiography for detection of nodules at a statistically significant level (p < .05). Discussion of the comparison study emphasizes the unique combination of data collection and analysis techniques employed, as well as the limitations of comparison techniques in the larger context of technology assessment.

Quick and simple estimation of bacteria using a fluorescent paracetamol dimer-Au nanoparticle composite

NASA Astrophysics Data System (ADS)

Sahoo, Amaresh Kumar; Sharma, Shilpa; Chattopadhyay, Arun; Ghosh, Siddhartha Sankar

2012-02-01

Rapid, simple and sensitive detection of bacterial contamination is critical for safeguarding public health and the environment. Herein, we report an easy method of detection as well as enumeration of the bacterial cell number on the basis of fluorescence quenching of a non-antibacterial fluorescent nanocomposite, consisting of paracetamol dimer (PD) and Au nanoparticles (NPs), in the presence of bacteria. The composite was synthesized by reaction of paracetamol (p-hydroxyacetanilide) with HAuCl4. The Au NPs of the composite were characterized using UV-Vis spectroscopy, transmission electron microscopy (TEM), X-ray diffraction and selected area electron diffraction analysis. The paracetamol dimer in the composite showed emission peak at 435 nm when excited at 320 nm. The method successfully detected six bacterial strains with a sensitivity of 100 CFU mL-1. The Gram-positive and Gram-negative bacteria quenched the fluorescence of the composite differently, making it possible to distinguish between the two. The TEM analysis showed interaction of the composite with bacteria without any apparent damage to the bacteria. The chi-square test established the accuracy of the method. Quick, non-specific and highly sensitive detection of bacteria over a broad range of logarithmic dilutions within a short span of time demonstrates the potential of this method as an alternative to conventional methods.Rapid, simple and sensitive detection of bacterial contamination is critical for safeguarding public health and the environment. Herein, we report an easy method of detection as well as enumeration of the bacterial cell number on the basis of fluorescence quenching of a non-antibacterial fluorescent nanocomposite, consisting of paracetamol dimer (PD) and Au nanoparticles (NPs), in the presence of bacteria. The composite was synthesized by reaction of paracetamol (p-hydroxyacetanilide) with HAuCl4. The Au NPs of the composite were characterized using UV-Vis spectroscopy, transmission electron microscopy (TEM), X-ray diffraction and selected area electron diffraction analysis. The paracetamol dimer in the composite showed emission peak at 435 nm when excited at 320 nm. The method successfully detected six bacterial strains with a sensitivity of 100 CFU mL-1. The Gram-positive and Gram-negative bacteria quenched the fluorescence of the composite differently, making it possible to distinguish between the two. The TEM analysis showed interaction of the composite with bacteria without any apparent damage to the bacteria. The chi-square test established the accuracy of the method. Quick, non-specific and highly sensitive detection of bacteria over a broad range of logarithmic dilutions within a short span of time demonstrates the potential of this method as an alternative to conventional methods. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr11837h

A colloidal gold nanoparticle-based immunochromatographic test strip for rapid and convenient detection of Staphylococcus aureus.

PubMed

Niu, Kaili; Zheng, Xiaoping; Huang, Chusen; Xul, Kuan; Zhi, Yuan; Shen, Hebai; Jia, Nengqin

2014-07-01

An immunochromatographic test strip using gold nanoparticles-staphylococcus aureus monoclonal antibody conjugates was developed for the rapid and convenient detection of staphylococcus aureus based on a double-antibody sandwich format. The detection limit and the detection rate of this test strip is 10(3) CFU /mL and 98.7%, respectively. It could be used for the rapid detection of staphylococcus aureus in food and the results can be visually identified by the naked eye within 10 min. Compared with conventional bacterial detection methods, this developed immunochromatographic assay based test strip has several advantages including simple, fast, low-cost, favorable sensitivity and specificity, exhibiting a great potential for application in food safety control systems and clinical diagnosis.

Optical Method for Detecting Displacements and Strains at Ultra-High Temperatures During Thermo-Mechanical Testing

NASA Technical Reports Server (NTRS)

Roth, Mark C. (Inventor); Smith, Russell W. (Inventor); Sikora, Joseph G. (Inventor); Rivers, H. Kevin (Inventor); Johnston, William M. (Inventor)

2016-01-01

An ultra-high temperature optical method incorporates speckle optics for sensing displacement and strain measurements well above conventional measurement techniques. High temperature pattern materials are used which can endure experimental high temperature environments while simultaneously having a minimum optical aberration. A purge medium is used to reduce or eliminate optical distortions and to reduce, and/or eliminate oxidation of the target specimen.

Prevalence of virulent Yersinia enterocolitica in bulk raw milk and retail cheese in northern-west of Iran.

PubMed

Hanifian, Shahram; Khani, Sajjad

2012-04-02

To determine the prevalence of virulent Yersinia enterocolitica, 554 samples consisting of 354 bulk raw milks and 200 traditional cheeses were collected from different parts of Eastern-Azerbaijan province, during a 23-month period from 2008 to 2010. The occurrence of virulent strains of Y. enterocolitica in samples enriched in peptone sorbitol bile broth (PSBB) was evaluated via the detection of attachment invasion locus (ail) gene by PCR. The viability of virulent Y. enterocolitica in the PCR-positive samples was tested using conventional culture method and the isolates were confirmed by the second-phase ail-PCR. According to the results, 8.66% of total samples including 7.62% of bulk raw milks and 10.5% of raw milk cheeses were found ail-positive by PCR method; subsequently Y. enterocolitica was isolated by the culture method and confirmed by the second phase ail-PCR in 2.88% of total samples including 2.26% of raw milks and 4% of cheese samples. It was concluded that, a sample enrichment followed by ail-PCR was more sensitive and robust to detect and distinguish the virulent strains of Y. enterocolitica compared to the conventional culture method. Copyright © 2012 Elsevier B.V. All rights reserved.

Culture versus PCR for Salmonella Species Identification in Some Dairy Products and Dairy Handlers with Special Concern to Its Zoonotic Importance.

PubMed

Gwida, Mayada M; Al-Ashmawy, Maha A M

2014-01-01

A total of 200 samples of milk and dairy products as well as 120 samples of dairy handlers were randomly collected from different dairy farms and supermarkets in Dakahlia Governorate, Egypt. The conventional cultural and serotyping methods for detection of Salmonella in dairy products were applied and the results were compared with those obtained by molecular screening assay using (ttr sequence). The obtained results revealed that 21% of milk and dairy products (42/200) were positive for Salmonella species using enrichment culture-based PCR method, while 12% of different dairy samples (24/200) were found to be positive for Salmonella species by using the conventional culture methods. Two stool specimens out of 40 apparently healthy dairy handlers were positive by the PCR method. Serotyping of Salmonella isolates revealed that 58.3% (14/24) from different dairy products were contaminated with Salmonella Typhimurium. We conclude that the enrichment culture-based PCR assay has high sensitivity and specificity for detection of Salmonella species in dairy products and handlers. High incidence of Salmonella Typhimurium in the examined dairy samples highlights the important role played by milk and dairy products as a vehicle in disease prevalence. Great effort should be applied for reducing foodborne risk for consumers.

Clinical evaluation of β-tubulin real-time PCR for rapid diagnosis of dermatophytosis, a comparison with mycological methods.

PubMed

Motamedi, Marjan; Mirhendi, Hossein; Zomorodian, Kamiar; Khodadadi, Hossein; Kharazi, Mahboobeh; Ghasemi, Zeinab; Shidfar, Mohammad Reza; Makimura, Koichi

2017-10-01

Following our previous report on evaluation of the beta tubulin real-time PCR for detection of dermatophytosis, this study aimed to compare the real-time PCR assay with conventional methods for the clinical assessment of its diagnostic performance. Samples from a total of 853 patients with suspected dermatophyte lesions were subjected to direct examination (all samples), culture (499 samples) and real-time PCR (all samples). Fungal DNA was extracted directly from clinical samples using a conical steel bullet, followed by purification with a commercial kit and subjected to the Taq-Man probe-based real-time PCR. The study showed that among the 499 specimens for which all three methods were used, 156 (31.2%), 128 (25.6%) and 205 (41.0%) were found to be positive by direct microscopy, culture and real-time PCR respectively. Real-time PCR significantly increased the detection rate of dermatophytes compared with microscopy (288 vs 229) with 87% concordance between the two methods. The sensitivity, specificity, positive predictive value, and negative predictive value of the real-time PCR was 87.5%, 85%, 66.5% and 95.2% respectively. Although real-time PCR performed better on skin than on nail samples, it should not yet fully replace conventional diagnosis. © 2017 Blackwell Verlag GmbH.

Development of sandwich dot-ELISA for specific detection of Ochratoxin A and its application on to contaminated cereal grains originating from India

PubMed Central

Venkataramana, M.; Rashmi, R.; Uppalapati, Siva R.; Chandranayaka, S.; Balakrishna, K.; Radhika, M.; Gupta, Vijai K.; Batra, H. V.

2015-01-01

In the present study, generation and characterization of a highly specific monoclonal antibody (mAb) against Ochratoxin A (OTA) was undertaken. The generated mAb was further used to develop a simple, fast, and sensitive sandwich dot-ELISA (s-dot ELISA) method for detection of OTA from contaminated food grain samples. The limit of detection (LOD) of the developed enzyme-linked immunosorbent assay (ELISA) method was determined as 5.0 ng/mL of OTA. Developed method was more specific toward OTA and no cross reactivity was observed with the other tested mycotoxins such as deoxynivalenol, fumonisin B1, or aflatoxin B1. To assess the utility and reliability of the developed method, several field samples of maize, wheat and rice (n = 195) collected from different geographical regions of southern Karnataka region of India were evaluated for the OTA occurrence. Seventy two out of 195 samples (19 maize, 38 wheat, and 15 rice) were found to be contaminated by OTA by s-dot ELISA. The assay results were further co-evaluated with conventional analytical high-performance liquid chromatography (HPLC) method. Results of the s-dot ELISA are in concordance with HPLC except for three samples that were negative for OTA presence by s-dot ELISA but found positive by HPLC. Although positive by HPLC, the amount of OTA in the three samples was found to be lesser than the accepted levels (>5 μg/kg) of OTA presence in cereals. Therefore, in conclusion, the developed s-dot ELISA is a better alternative for routine cereal based food and feed analysis in diagnostic labs to check the presence of OTA over existing conventional culture based, tedious analytical methods. PMID:26074899

Quantitative real-time PCR as a sensitive protein-protein interaction quantification method and a partial solution for non-accessible autoactivator and false-negative molecule analysis in the yeast two-hybrid system.

PubMed

Maier, Richard H; Maier, Christina J; Hintner, Helmut; Bauer, Johann W; Onder, Kamil

2012-12-01

Many functional proteomic experiments make use of high-throughput technologies such as mass spectrometry combined with two-dimensional polyacrylamide gel electrophoresis and the yeast two-hybrid (Y2H) system. Currently there are even automated versions of the Y2H system available that can be used for proteome-wide research. The Y2H system has the capacity to deliver a profusion of Y2H positive colonies from a single library screen. However, subsequent analysis of these numerous primary candidates with complementary methods can be overwhelming. Therefore, a method to select the most promising candidates with strong interaction properties might be useful to reduce the number of candidates requiring further analysis. The method described here offers a new way of quantifying and rating the performance of positive Y2H candidates. The novelty lies in the detection and measurement of mRNA expression instead of proteins or conventional Y2H genetic reporters. This method correlates well with the direct genetic reporter readouts usually used in the Y2H system, and has greater sensitivity for detecting and quantifying protein-protein interactions (PPIs) than the conventional Y2H system, as demonstrated by detection of the Y2H false-negative PPI of RXR/PPARG. Approximately 20% of all proteins are not suitable for the Y2H system, the so-called autoactivators. A further advantage of this method is the possibility to evaluate molecules that usually cannot be analyzed in the Y2H system, exemplified by a VDR-LXXLL motif peptide interaction. Copyright © 2012 Elsevier Inc. All rights reserved.

Bacteriophage Amplification-Coupled Detection and Identification of Bacterial Pathogens

NASA Astrophysics Data System (ADS)

Cox, Christopher R.; Voorhees, Kent J.

Current methods of species-specific bacterial detection and identification are complex, time-consuming, and often require expensive specialized equipment and highly trained personnel. Numerous biochemical and genotypic identification methods have been applied to bacterial characterization, but all rely on tedious microbiological culturing practices and/or costly sequencing protocols which render them impractical for deployment as rapid, cost-effective point-of-care or field detection and identification methods. With a view towards addressing these shortcomings, we have exploited the evolutionarily conserved interactions between a bacteriophage (phage) and its bacterial host to develop species-specific detection methods. Phage amplification-coupled matrix assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF-MS) was utilized to rapidly detect phage propagation resulting from species-specific in vitro bacterial infection. This novel signal amplification method allowed for bacterial detection and identification in as little as 2 h, and when combined with disulfide bond reduction methods developed in our laboratory to enhance MALDI-TOF-MS resolution, was observed to lower the limit of detection by several orders of magnitude over conventional spectroscopy and phage typing methods. Phage amplification has been combined with lateral flow immunochromatography (LFI) to develop rapid, easy-to-operate, portable, species-specific point-of-care (POC) detection devices. Prototype LFI detectors have been developed and characterized for Yersinia pestis and Bacillus anthracis, the etiologic agents of plague and anthrax, respectively. Comparable sensitivity and rapidity was observed when phage amplification was adapted to a species-specific handheld LFI detector, thus allowing for rapid, simple, POC bacterial detection and identification while eliminating the need for bacterial culturing or DNA isolation and amplification techniques.

Identification of suitable sites for mountain ginseng cultivation using GIS and geo-temperature.

PubMed

Kang, Hag Mo; Choi, Soo Im; Kim, Hyun

2016-01-01

This study was conducted to explore an accurate site identification technique using a geographic information system (GIS) and geo-temperature (gT) for locating suitable sites for growing cultivated mountain ginseng (CMG; Panax ginseng), which is highly sensitive to the environmental conditions in which it grows. The study site was Jinan-gun, South Korea. The spatial resolution for geographic data was set at 10 m × 10 m, and the temperatures for various climatic factors influencing CMG growth were calculated by averaging the 3-year temperatures obtained from the automatic weather stations of the Korea Meteorological Administration. Identification of suitable sites for CMG cultivation was undertaken using both a conventional method and a new method, in which the gT was added as one of the most important factors for crop cultivation. The results yielded by the 2 methods were then compared. When the gT was added as an additional factor (new method), the proportion of suitable sites identified decreased by 0.4 % compared with the conventional method. However, the proportion matching real CMG cultivation sites increased by 3.5 %. Moreover, only 68.2 % corresponded with suitable sites identified using the conventional factors; i.e., 31.8 % were newly detected suitable sites. The accuracy of GIS-based identification of suitable CMG cultivation sites improved by applying the temperature factor (i.e., gT) in addition to the conventionally used factors.

Helical CT scan with 2D and 3D reconstructions and virtual endoscopy versus conventional endoscopy in the assessment of airway disease in neonates, infants and children.

PubMed

Yunus, Mahira

2012-11-01

To study the use of helical computed tomography 2-D and 3-D images, and virtual endoscopy in the evaluation of airway disease in neonates, infants and children and its value in lesion detection, characterisation and extension. Conducted at Al-Noor Hospital, Makkah, Saudi Arabia, from January 1 to June 30, 2006, the study comprised of 40 patients with strider, having various causes of airway obstruction. They were examined by helical CT scan with 2-D and 3-D reconstructions and virtual endoscopy. The level and characterisation of lesions were carried out and results were compared with actual endoscopic findings. Conventional endoscopy was chosen as the gold standard, and the evaluation of endoscopy was done in terms of sensitivity and specificity of the procedure. For statistical purposes, SPSS version 10 was used. All CT methods detected airway stenosis or obstruction. Accuracy was 98% (n=40) for virtual endoscopy, 96% (n=48) for 3-D external rendering, 90% (n=45) for multiplanar reconstructions and 86% (n=43) for axial images. Comparing the results of 3-D internal and external volume rendering images with conventional endoscopy for detection and grading of stenosis were closer than with 2-D minimum intensity multiplanar reconstruction and axial CT slices. Even high-grade stenosis could be evaluated with virtual endoscope through which conventional endoscope cannot be passed. A case of 4-year-old patient with tracheomalacia could not be diagnosed by helical CT scan and virtual bronchoscopy which was diagriosed on conventional endoscopy and needed CT scan in inspiration and expiration. Virtual endoscopy [VE] enabled better assessment of stenosis compared to the reading of 3-D external rendering, 2-D multiplanar reconstruction [MPR] or axial slices. It can replace conventional endoscopy in the assessment of airway disease without any additional risk.