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Sample records for fingerstick blood samples

  1. Whole genome transcript profiling from fingerstick blood samples: a comparison and feasibility study

    PubMed Central

    2009-01-01

    Background Whole genome gene expression profiling has revolutionized research in the past decade especially with the advent of microarrays. Recently, there have been significant improvements in whole blood RNA isolation techniques which, through stabilization of RNA at the time of sample collection, avoid bias and artifacts introduced during sample handling. Despite these improvements, current human whole blood RNA stabilization/isolation kits are limited by the requirement of a venous blood sample of at least 2.5 mL. While fingerstick blood collection has been used for many different assays, there has yet to be a kit developed to isolate high quality RNA for use in gene expression studies from such small human samples. The clinical and field testing advantages of obtaining reliable and reproducible gene expression data from a fingerstick are many; it is less invasive, time saving, more mobile, and eliminates the need of a trained phlebotomist. Furthermore, this method could also be employed in small animal studies, i.e. mice, where larger sample collections often require sacrificing the animal. In this study, we offer a rapid and simple method to extract sufficient amounts of high quality total RNA from approximately 70 μl of whole blood collected via a fingerstick using a modified protocol of the commercially available Qiagen PAXgene RNA Blood Kit. Results From two sets of fingerstick collections, about 70 uL whole blood collected via finger lancet and capillary tube, we recovered an average of 252.6 ng total RNA with an average RIN of 9.3. The post-amplification yields for 50 ng of total RNA averaged at 7.0 ug cDNA. The cDNA hybridized to Affymetrix HG-U133 Plus 2.0 GeneChips had an average % Present call of 52.5%. Both fingerstick collections were highly correlated with r2 values ranging from 0.94 to 0.97. Similarly both fingerstick collections were highly correlated to the venous collection with r2 values ranging from 0.88 to 0.96 for fingerstick collection 1

  2. A novel device for collecting and dispensing fingerstick blood for point of care testing.

    PubMed

    Sauer-Budge, Alexis F; Brookfield, Samuel J; Janzen, Ronald; McGray, Sarah; Boardman, Anna; Wirz, Holger; Pollock, Nira R

    2017-01-01

    The increased world-wide availability of point-of-care (POC) tests utilizing fingerstick blood has led to testing scenarios in which multiple separate fingersticks are performed during a single patient encounter, generating cumulative discomfort and reducing testing efficiency. We have developed a device capable of a) collection of up to 100 μL of fingerstick blood from a single fingerstick by capillary action, and b) dispensing this blood in variable increments set by the user. We tested the prototype device both in a controlled laboratory setting and in a fingerstick study involving naive device users, and found it to have accuracy and precision similar to a conventional pipettor. The users also found the device to be easy to use, and recommended minor ergonomic improvements. Our device would allow performance of multiple POC tests from a single fingerstick blood sample, thus providing a novel functionality that may be of use in many testing settings worldwide.

  3. A novel device for collecting and dispensing fingerstick blood for point of care testing

    PubMed Central

    Janzen, Ronald; McGray, Sarah; Boardman, Anna; Wirz, Holger

    2017-01-01

    The increased world-wide availability of point-of-care (POC) tests utilizing fingerstick blood has led to testing scenarios in which multiple separate fingersticks are performed during a single patient encounter, generating cumulative discomfort and reducing testing efficiency. We have developed a device capable of a) collection of up to 100 μL of fingerstick blood from a single fingerstick by capillary action, and b) dispensing this blood in variable increments set by the user. We tested the prototype device both in a controlled laboratory setting and in a fingerstick study involving naive device users, and found it to have accuracy and precision similar to a conventional pipettor. The users also found the device to be easy to use, and recommended minor ergonomic improvements. Our device would allow performance of multiple POC tests from a single fingerstick blood sample, thus providing a novel functionality that may be of use in many testing settings worldwide. PMID:28837616

  4. Blood cholesterol screening in several environments using a portable, dry-chemistry analyzer and fingerstick blood samples. Lipid Research Clinics Cholesterol Screening Study Group.

    PubMed

    Bradford, R H; Bachorik, P S; Roberts, K; Williams, O D; Gotto, A M

    1990-01-01

    A multicenter study of blood cholesterol screening was performed in several typical environments, such as community sites (shopping malls and a supermarket), health care sites, work sites, a blood bank and a school. Cholesterol was measured with a portable, dry-chemistry analyzer using capillary blood obtained by fingerstick. Data are reported from a total of 13,824 participants, spanning the entire age spectrum. Overall, 25% of screened subjects had blood cholesterol levels above the age-specific cutpoints used in the current study. Although in the aggregate this screening experience very closely approximates the expected level of referrals, the proportion of referred screened subjects differed significantly among the 5 types of screening environments and by gender. Follow-up telephone interviews indicated that 53% of referrals had initiated a physician contact. More than 75% of those who had seen a physician reported that the diagnosis of hypercholesterolemia had been confirmed, and almost 72% had been prescribed a diet. A large proportion of referred screened subjects reported having modified their diet, particularly when recommended to do so by a physician. This study has yielded encouraging evidence that physicians gave referred screened subjects appropriate initial advice for managing hypercholesterolemia. The new technology for blood cholesterol measurement evaluated in the current study has proven to be a feasible and reliable means for measuring blood cholesterol in typical screening settings.

  5. The use of finger-stick blood to assess lactate in critically ill surgical patients.

    PubMed

    Sabat, Joseph; Gould, Scott; Gillego, Ezra; Hariprashad, Anita; Wiest, Christine; Almonte, Shailyn; Lucido, David J; Gave, Asaf; Leitman, I Michael; Eiref, Simon D

    2016-09-01

    Using finger-stick capillary blood to assess lactate from the microcirculation may have utility in treating critically ill patients. Our goals were to determine how finger-stick capillary lactate correlates with arterial lactate levels in patients from the surgical intensive care unit, and to compare how capillary and arterial lactate trend over time in patients undergoing resuscitation for shock. Capillary whole blood specimens were obtained from finger-sticks using a lancet, and assessed for lactate via a handheld point-of-care device as part of an "investigational use only" study. Comparison was made to arterial blood specimens that were assessed for lactate by standard laboratory reference methods. 40 patients (mean age 68, mean APACHEII 18, vasopressor use 62%) were included. The correlation between capillary and arterial lactate levels was 0.94 (p < 0.001). Capillary lactate measured slightly higher on average than paired arterial values, with a mean difference 0.99 mmol/L. In patients being resuscitated for septic and hemorrhagic shock, capillary and arterial lactate trended closely over time: rising, peaking, and falling in tandem. Clearance of capillary and arterial lactate mirrored clinical improvement, normalizing in all patients except two that expired. Finger-stick capillary lactate both correlates and trends closely with arterial lactate in critically ill surgical patients, undergoing resuscitation for shock.

  6. Association of δ¹³C in fingerstick blood with added-sugar and sugar-sweetened beverage intake.

    PubMed

    Davy, Brenda M; Jahren, A Hope; Hedrick, Valisa E; Comber, Dana L

    2011-06-01

    A reliance on self-reported dietary intake measures is a common research limitation, thus the need for dietary biomarkers. Added-sugar intake may play a role in the development and progression of obesity and related comorbidities; common sweeteners include corn and sugar cane derivatives. These plants contain a high amount of ¹³C, a naturally occurring stable carbon isotope. Consumption of these sweeteners, of which sugar-sweetened beverages are the primary dietary source, might be reflected in the δ¹³C value of blood. Fingerstick blood represents an ideal substrate for bioassay because of its ease of acquisition. The objective of this investigation was to determine if the δ¹³C value of fingerstick blood is a potential biomarker of added-sugar and sugar-sweetened beverage intake. Individuals aged 21 years and older (n = 60) were recruited to attend three laboratory visits; assessments completed at each visit depended upon a randomly assigned sequence (sequence one or two). The initial visit included assessment of height, weight, and dietary intake (sequence one: beverage intake questionnaire, sequence two: 4-day food intake record). Sequence one participants completed a food intake record at visit two, and nonfasting blood samples were obtained via routine fingersticks at visits one and three. Sequence two participants completed a beverage intake questionnaire at visit two, and provided fingerstick blood samples at visits two and three. Samples were analyzed for δ¹³C value using natural abundance stable isotope mass spectrometry. δ¹³C value was compared to dietary outcomes in all participants, as well as among those in the highest and lowest tertile of added-sugar intake. Reported mean added-sugar consumption was 66 ± 5 g/day, and sugar-sweetened beverage consumption was 330 ± 53 g/day and 134 ± 25 kcal/day. Mean fingerstick δ¹³C value was -19.94‰ ± 0.10‰, which differed by body mass index status. δ¹³C value was associated (all P < 0

  7. The difference between fingerstick and venous hemoglobin and hematocrit varies by sex and iron stores.

    PubMed

    Cable, Ritchard G; Steele, Whitney R; Melmed, Russell S; Johnson, Bryce; Mast, Alan E; Carey, Patricia M; Kiss, Joseph E; Kleinman, Steven H; Wright, David J

    2012-05-01

    Fingerstick blood samples are used to estimate donor venous hemoglobin (Hb). Fingerstick Hb or hematocrit (Hct) was determined routinely for 2425 selected donors at six blood centers, along with venous Hb. Using sex and measures of iron status including absent iron stores (AIS; ferritin < 12 ng/mL), linear regression models were developed to predict venous Hb from fingerstick. Across all subjects, fingerstick Hb was higher than venous Hb in the higher part of the clinical range, but lower in the lower part of the range. The relationship varied by sex and iron status. Across centers, a female donor had on average a venous Hb result 0.5 to 0.8 g/dL lower than a male donor with the same fingerstick Hb and iron status. Similarly, a donor with AIS had on average a venous Hb result 0.3 to 1.1 g/dL lower than an iron-replete donor with the same fingerstick value and sex. An iron-replete male donor with a fingerstick result at the cutoff (Hb 12.5 g/dL) had an acceptable expected venous Hb (12.8 to 13.8 g/dL). A female donor with AIS with a fingerstick result at the cutoff had an expected venous Hb below 12.5 g/dL (11.7 to 12.4 g/dL). Of females with AIS, 40.2% donated blood when their venous Hb was less than 12.5 g/dL. Fingerstick is considered a useful estimator of venous Hb. However, in some donor groups, particularly female donors with AIS, fingerstick overestimates venous Hb at the donation cutoff. This significant limitation should be considered in setting donor fingerstick Hb or Hct requirements. © 2011 American Association of Blood Banks.

  8. AccuStat whole blood fingerstick test for Helicobacter pylori infection: a reliable screening method.

    PubMed

    Harrison, J R; Bevan, J; Furth, E E; Metz, D C

    1998-07-01

    Helicobacter pylori antibody testing is accurate for diagnosing untreated patients. Rapid serum testing is as accurate as formal enzyme-linked immunosorbent assay (ELISA) testing. As whole blood fingerstick tests may become the diagnostic method of choice if they are of similar accuracy, 51 patients were studied who had not taken antibiotics, bismuth, sucralfate, or proton pump inhibitors. Concordance between C-14 Urea Breath Testing and HM-CAP ELISA testing served as the study standard for H. pylori diagnosis. Rapid antibody testing was performed with the AccuStat whole blood (Boehringer Mannheim, Mannheim, Germany) and FlexSure HP (Smith Kline Diagnostics, San Jose, CA) serum tests. Antral biopsy for CLO testing and histological evaluation with thiazine staining were available for 18 (35.3%) and 20 patients (39.2%), respectively. Nineteen of 50 patients (38%) were infected. (One patient had discordant tests and was excluded.) FlexSure HP and AccuStat were each positive in 18 (36%) and 19 patients (38%) with sensitivity, specificity, and positive and negative predictive values of 89.5% and 89.5%, 96.8% and 93.5%, 94.4% and 89.5%, and 93.8% and 93.5%, respectively. There were two false-negative FlexSure HP and AccuStat tests and three false-positive tests--1 FlexSure and 2 AccuStat results. CLO test and histology concurred in every case tested. We conclude that both rapid antibody tests are accurate and suitable for screening patients not previously treated for H. pylori infection. Since the AccuStat has preserved diagnostic strength, is less costly, takes less time, and is less labor intensive, whole blood testing is the screening test of choice.

  9. The difference between fingerstick and venous hemoglobin and hematocrit varies by sex and iron stores

    PubMed Central

    Cable, Ritchard G.; Steele, Whitney R.; Melmed, Russell S.; Johnson, Bryce; Mast, Alan E.; Carey, Patricia M.; Kiss, Joseph E.; Kleinman, Steven H.; Wright, David J.

    2013-01-01

    BACKGROUND Fingerstick blood samples are used to estimate donor venous hemoglobin (Hb). STUDY DESIGN AND METHODS Fingerstick Hb or hematocrit (Hct) was determined routinely for 2425 selected donors at six blood centers, along with venous Hb. Using sex and measures of iron status including absent iron stores (AIS; ferritin < 12 ng/mL), linear regression models were developed to predict venous Hb from fingerstick. RESULTS Across all subjects, fingerstick Hb was higher than venous Hb in the higher part of the clinical range, but lower in the lower part of the range. The relationship varied by sex and iron status. Across centers, a female donor had on average a venous Hb result 0.5 to 0.8 g/dL lower than a male donor with the same fingerstick Hb and iron status. Similarly, a donor with AIS had on average a venous Hb result 0.3 to 1.1 g/dL lower than an iron-replete donor with the same fingerstick value and sex. An iron-replete male donor with a fingerstick result at the cutoff (Hb 12.5 g/dL) had an acceptable expected venous Hb (12.8 to 13.8 g/dL). A female donor with AIS with a fingerstick result at the cutoff had an expected venous Hb below 12.5 g/dL (11.7 to 12.4 g/dL). Of females with AIS, 40.2% donated blood when their venous Hb was less than 12.5 g/dL. CONCLUSIONS Fingerstick is considered a useful estimator of venous Hb. However, in some donor groups, particularly female donors with AIS, fingerstick overestimates venous Hb at the donation cutoff. This significant limitation should be considered in setting donor fingerstick Hb or Hct requirements. PMID:22014071

  10. Sensitivity of Five Rapid HIV Tests on Oral Fluid or Finger-Stick Whole Blood: A Real-Time Comparison in a Healthcare Setting

    PubMed Central

    Pavie, Juliette; Rachline, Anne; Loze, Bénédicte; Niedbalski, Laurence; Delaugerre, Constance; Laforgerie, Eric; Plantier, Jean-Christophe; Rozenbaum, Willy; Chevret, Sylvie; Molina, Jean-Michel; Simon, François

    2010-01-01

    Background Health authorities in several countries recently recommended the expansion of human immunodeficiency virus (HIV) antibody testing, including the use of rapid tests. Several HIV rapid tests are now licensed in Europe but their sensitivity on total blood and/or oral fluid in routine healthcare settings is not known. Methods and Findings 200 adults with documented HIV-1 (n = 194) or HIV-2 infection (n = 6) were prospectively screened with five HIV rapid tests using either oral fluid (OF) or finger-stick whole blood (FSB). The OraQuick Advance rapid HIV1/2® was first applied to OF and then to FSB, while the other tests were applied to FSB, in the following order: Vikia HIV 1/2®, Determine HIV 1–2®, Determine® HIV-1/2 Ag/Ab Combo® and INSTI HIV-1/HIV-2®. Tests negative on FSB were repeated on paired serum samples. Twenty randomly selected HIV-seronegative subjects served as controls, and the results were read blindly. Most patients had HIV-1 subtype B infection (63.3%) and most were on antiretroviral therapy (68.5%). Sensitivity was 86.5%, 94.5%, 98.5%, 94.9%, 95.8% and 99% respectively, with OraQuick OF, OraQuick FSB, Vikia, Determine, Determine Ag/Ab Combo and INSTI (p<0.0001). OraQuick was less sensitive on OF than on FSB (p = 0.008). Among the six patients with three or more negative tests, two had recent HIV infection and four patients on antiretroviral therapy had undetectable plasma viral load. When patients positive in all the tests were compared with patients who had at least one negative test, only a plasma HIV RNA level <200 cp/ml was significantly associated with a false-negative result (p = 0.009). When the 33 rapid tests negative on FSB were repeated on serum, all but six (5 negative with OraQuick, 1 with INSTI) were positive. The sensitivity of OraQuick, Determine and Determine Ag/Ab Combo was significantly better on serum than on FSB (97.5%, p = 0.04; 100%, p = 0.004; and 100%, p = 0.02, respectively

  11. A comparison of patient acceptance of fingerstick whole blood and oral fluid rapid HIV screening in an emergency department.

    PubMed

    White, Douglas A E; Scribner, Alicia N; Huang, Jennifer V

    2009-09-01

    Although whole blood rapid HIV testing has a greater sensitivity and specificity compared with oral fluid (OF) testing, patients prefer HIV testing using OF specimen collection. Whether patient preference for noninvasive collection methods affects acceptance of HIV screening in clinical practice, however, is unknown. To determine whether patient acceptance of fingerstick whole blood (FWB) and OF rapid HIV screening differs in an emergency department setting. From May 1 to June 30, 2007, triage-based testers offered rapid HIV screening to patients. OF and FWB tests were available on alternate days. Eligible patients were medically stable, > or =15 years of age, and not known to be HIV infected. : Two thousand two hundred one patients were referred for HIV screening: 1089 on OF days and 1112 on FWB screening days. Screening rates with OF and FWB were similar (61.9% vs. 59.1%, P = 0.18). Although most reasons why patients declined screening were similar for the groups, the specimen collection method was the primary reason for refusal by 25 patients who declined FWB screening compared with none of the patients who declined OF screening. Among emergency department patients, the preference for 1 rapid test collection modality over another has a minimal effect on actual screening rates.

  12. Measures of Viral Load using Abbott Real-Time HIV-1 Assay on Venous and Fingerstick Dried Blood Spots from Provider-Collected Specimens in Malawian District Hospitals

    PubMed Central

    Rutstein, Sarah E.; Kamwendo, Deborah; Lugali, Lebah; Thengolose, Isaac; Tegha, Gerald; Fiscus, Susan A.; Nelson, Julie A. E.; Hosseinipour, Mina C.; Sarr, Abdoulaye; Gupta, Sundeep; Chimbwandira, Frank; Mwenda, Reuben; Mataya, Ronald

    2014-01-01

    Background Viral suppression is a key indicator of antiretroviral therapy (ART) response among HIV-infected patients. Dried blood spots (DBS) are an appealing alternative to conventional plasma-based virologic testing, improving access to monitoring in resource-limited settings. However, validity of DBS obtained from fingerstick in field settings remains unknown. Objectives Investigate feasibility and accuracy of DBS vs plasma collected by healthcare workers in real-world settings of remote hospitals in Malawi. Compare venous DBS to fingerstick DBS for identifying treatment failure. Study design We recruited patients from ART clinics at two district hospitals in Malawi, collecting plasma, venous DBS (vDBS), and fingerstick DBS (fsDBS) cards for the first 149 patients, and vDBS and fsDBS only for the subsequent 398 patients. Specimens were tested using Abbott RealTime HIV-1 Assay (lower detection limit 40 copies/ml (plasma) and 550 copies/ml (DBS)). Results 21/149 (14.1%) had detectable viremia (>1.6 log copies/ml), 13 of which were detectable for plasma, vDBS, and fsDBS. Linear regression demonstrated high correlation for plasma vs. DBS (vDBS: β=1.19, R2 0.93 (p<0.0001); fsDBS β=1.20, R2 0.90 (p<0.0001)) and vDBS vs. fsDBS (β=0.88, R2 0.73, (p<0.0001)). Mean difference between plasma and vDBS was 0.51 log copies/ml [SD: 0.33] and plasma and fsDBS 0.46 log copies/ml [SD: 0.30]. At 5000 copies/ml, sensitivity was 100%, and specificity was 98.6% and 97.8% for vDBS and fsDBS, respectively, compared to plasma. Conclusions DBS from venipuncture and fingerstick perform well at the failure threshold of 5000 copies/ml. Fingerstick specimen source may improve access to virologic treatment monitoring in resource-limited settings given task-shifting in high-volume, low-resource facilities. PMID:24906641

  13. A Finger-Stick Whole-Blood HIV Self-Test as an HIV Screening Tool Adapted to the General Public.

    PubMed

    Prazuck, Thierry; Karon, Stephen; Gubavu, Camelia; Andre, Jerome; Legall, Jean Marie; Bouvet, Elisabeth; Kreplak, Georges; Teglas, Jean Paul; Pialoux, Gilles

    2016-01-01

    In 2013, the French Health Authority approved the use of HIV self-tests in pharmacies for the general public. This screening tool will allow an increase in the number of screenings and a reduction in the delay between infection and diagnosis, thus reducing the risk of further infections. We previously compared 5 HIV-self test candidates (4 oral fluid and one whole blood) and demonstrated that the whole blood HIV test exhibited the optimal level of performance (sensitivity/specificity). We studied the practicability of an easy-to-use finger-stick whole blood HIV self-test "autotest VIH®", when used in the general public. This multicenter cross-sectional study involved 411 participants from the Parisian region (AIDES and HF association) between April and July 2014 and was divided into 2 separate studies: one evaluating the capability of participants to obtain an interpretable result using only the information notice, and a second evaluating the interpretation of test results, using a provided chart. A total of 411 consenting participants, 264 in the first study and 147 in the second, were included. All participants were over 18 years of age. In the first study, 99.2% of the 264 participants correctly administered the auto-test, and 21.2% needed, upon their request, telephone assistance. Ninety-two percent of participants responded that the test was easy/very easy to perform, and 93.5% did not find any difficulty obtaining a sufficient good quantity of blood. In the second study, 98.1% of the 147 participants correctly interpreted the results. The reading/interpretation errors concerned the negative (2.1%) or the indeterminate (3.3%) auto-tests. The success rate of handling and interpretation of this self-test is very satisfactory, demonstrating its potential for use by the general public and its utility to increase the number of opportunities to detect HIV patients.

  14. A Finger-Stick Whole-Blood HIV Self-Test as an HIV Screening Tool Adapted to the General Public

    PubMed Central

    Prazuck, Thierry; Karon, Stephen; Gubavu, Camelia; Andre, Jerome; Legall, Jean Marie; Bouvet, Elisabeth; Kreplak, Georges; Teglas, Jean Paul; Pialoux, Gilles

    2016-01-01

    Background In 2013, the French Health Authority approved the use of HIV self-tests in pharmacies for the general public. This screening tool will allow an increase in the number of screenings and a reduction in the delay between infection and diagnosis, thus reducing the risk of further infections. We previously compared 5 HIV-self test candidates (4 oral fluid and one whole blood) and demonstrated that the whole blood HIV test exhibited the optimal level of performance (sensitivity/specificity). We studied the practicability of an easy-to-use finger-stick whole blood HIV self-test “autotest VIH®”, when used in the general public. Methods and Materials This multicenter cross-sectional study involved 411 participants from the Parisian region (AIDES and HF association) between April and July 2014 and was divided into 2 separate studies: one evaluating the capability of participants to obtain an interpretable result using only the information notice, and a second evaluating the interpretation of test results, using a provided chart. Results A total of 411 consenting participants, 264 in the first study and 147 in the second, were included. All participants were over 18 years of age. In the first study, 99.2% of the 264 participants correctly administered the auto-test, and 21.2% needed, upon their request, telephone assistance. Ninety-two percent of participants responded that the test was easy/very easy to perform, and 93.5% did not find any difficulty obtaining a sufficient good quantity of blood. In the second study, 98.1% of the 147 participants correctly interpreted the results. The reading/interpretation errors concerned the negative (2.1%) or the indeterminate (3.3%) auto-tests. Conclusions The success rate of handling and interpretation of this self-test is very satisfactory, demonstrating its potential for use by the general public and its utility to increase the number of opportunities to detect HIV patients. PMID:26882229

  15. A Clinical Evaluation of Routine Blood Sampling Practices in Patients With Diabetes

    PubMed Central

    Pineau, Mitchel; Pynes, Mary Kate; Katz, Laurence B.; Ginsberg, Barry

    2014-01-01

    Background: There is a perception that patients with diabetes struggle to produce sufficient blood to fill glucose test strips, including strips with 1-µL fill requirements. The purpose of this study was to determine the volume of blood expressed when these patients perform routine fingersticks using their own lancing device and sampling technique and to evaluate the relationship between blood volume and pain. Methods: Sixty-four patients (type 1 or type 2 diabetes) performed 8 fingersticks using their own lancing device and preferred depth setting and lancing technique. Eight different commercially available lancing systems were used (8 patients/system). Blood volume and perceived pain were recorded after each fingerstick. Results: The mean blood volume across all patients was 3.1 µL (512 fingersticks), with 97% of patients expressing a mean of ≥1.0 µL of blood. There was no correlation between pain response and the volume of blood expressed. Nearly all patients agreed that they could easily and comfortably obtain a 1-µL blood sample, and most patients actually preferred a larger drop size to ease sampling and avoid wasting strips. Conclusion: These results provide evidence across 8 lancing systems that challenge the current perceptions that patients with diabetes struggle to produce sufficient blood samples to fill most test strips, including those with 1-µL fill requirements, and that obtaining larger volumes of blood is more painful. These results are consistent with the previous literature suggesting that patients derive no real benefits from very low strip volumes and generally prefer a blood drop size that enables them to confidently fill their test strip. PMID:24876439

  16. Immune Blood Sample Draw

    NASA Image and Video Library

    2012-04-26

    ISS030-E-257690 (26 April 2012) --- European Space Agency astronaut Andre Kuipers, Expedition 30 flight engineer, prepares for IMMUNE venous blood sample draws in the Columbus laboratory of the International Space Station. Following the blood draws, the samples were temporarily stowed in the Minus Eighty Laboratory Freezer for ISS 1 (MELFI-1) and later packed together with saliva samples on the Soyuz TMA-22 for return to Earth for analysis.

  17. Comparison of POCT and central laboratory blood glucose results using arterial, capillary, and venous samples from MICU patients on a tight glycemic protocol.

    PubMed

    Petersen, John R; Graves, Donna F; Tacker, Danyel H; Okorodudu, Anthony O; Mohammad, Amin A; Cardenas, Victor J

    2008-10-01

    Point of care (POC) glucose meters are routinely used to monitor glucose levels for patients on tight glycemic control therapy. We determined if glucose values were different for a POC glucose meter as compared to the main clinical laboratory for medical intensive care unit patients on a tight glycemic protocol and whether the site of blood sampling had a significant impact on glucose values. Eighty-four patients (114 paired samples) who were on a tight glycemic protocol in the period November 2005 through August 2006 were enrolled. After simultaneous blood draws, we compared the glucose levels for the glucose meter (arterial/venous/capillary), blood gas (arterial/venous), and central clinical laboratory (serum/plasma from arterial/venous samples). The mean glucose levels of all arterial/venous/fingerstick samples using the glucose meter demonstrated a positive bias of 0.7-0.9 mmol/l (12.6-16.2 mg/dl) (p<0.001) relative to central laboratory venous plasma. There was also a smaller positive (0.1-0.3 mmol/l or 1.8-5.4 mg/dl, p<0.05) bias for arterial/venous blood gas samples and laboratory arterial serum/plasma glucose samples. Using Parkes error grid analysis we were able to show that the bias for arterial or venous POC glucose results would have not impacted clinical care. This was not the case, however, for fingerstick sampling where a high bias could have significantly impacted clinical care. Additionally, in 3 fingerstick samples a severe underestimation (<46% of the central laboratory plasma result) was found. Glucose meters using arterial/venous whole blood may be utilized in the MICU; however, due to the increased variability of results we do not recommend the routine use of capillary blood sampling for monitoring glucose levels in the MICU setting.

  18. Field evaluation of a prototype paper-based point-of-care fingerstick transaminase test.

    PubMed

    Pollock, Nira R; McGray, Sarah; Colby, Donn J; Noubary, Farzad; Nguyen, Huyen; Nguyen, The Anh; Khormaee, Sariah; Jain, Sidhartha; Hawkins, Kenneth; Kumar, Shailendra; Rolland, Jason P; Beattie, Patrick D; Chau, Nguyen V; Quang, Vo M; Barfield, Cori; Tietje, Kathy; Steele, Matt; Weigl, Bernhard H

    2013-01-01

    Monitoring for drug-induced liver injury (DILI) via serial transaminase measurements in patients on potentially hepatotoxic medications (e.g., for HIV and tuberculosis) is routine in resource-rich nations, but often unavailable in resource-limited settings. Towards enabling universal access to affordable point-of-care (POC) screening for DILI, we have performed the first field evaluation of a paper-based, microfluidic fingerstick test for rapid, semi-quantitative, visual measurement of blood alanine aminotransferase (ALT). Our objectives were to assess operational feasibility, inter-operator variability, lot variability, device failure rate, and accuracy, to inform device modification for further field testing. The paper-based ALT test was performed at POC on fingerstick samples from 600 outpatients receiving HIV treatment in Vietnam. Results, read independently by two clinic nurses, were compared with gold-standard automated (Roche Cobas) results from venipuncture samples obtained in parallel. Two device lots were used sequentially. We demonstrated high inter-operator agreement, with 96.3% (95% C.I., 94.3-97.7%) agreement in placing visual results into clinically-defined "bins" (<3x, 3-5x, and >5x upper limit of normal), >90% agreement in validity determination, and intraclass correlation coefficient of 0.89 (95% C.I., 0.87-0.91). Lot variability was observed in % invalids due to hemolysis (21.1% for Lot 1, 1.6% for Lot 2) and correlated with lots of incorporated plasma separation membranes. Invalid rates <1% were observed for all other device controls. Overall bin placement accuracy for the two readers was 84% (84.3%/83.6%). Our findings of extremely high inter-operator agreement for visual reading-obtained in a target clinical environment, as performed by local practitioners-indicate that the device operation and reading process is feasible and reproducible. Bin placement accuracy and lot-to-lot variability data identified specific targets for device

  19. Use of Dried Capillary Blood Sampling for Islet Autoantibody Screening in Relatives: A Feasibility Study.

    PubMed

    Bingley, Polly J; Rafkin, Lisa E; Matheson, Della; Steck, Andrea K; Yu, Liping; Henderson, Courtney; Beam, Craig A; Boulware, David C

    2015-12-01

    Islet autoantibody testing provides the basis for assessment of risk of progression to type 1 diabetes. We set out to determine the feasibility and acceptability of dried capillary blood spot-based screening to identify islet autoantibody-positive relatives potentially eligible for inclusion in prevention trials. Dried blood spot (DBS) and venous samples were collected from 229 relatives participating in the TrialNet Pathway to Prevention Study. Both samples were tested for glutamic acid decarboxylase, islet antigen 2, and zinc transporter 8 autoantibodies, and venous samples were additionally tested for insulin autoantibodies and islet cell antibodies. We defined multiple autoantibody positive as two or more autoantibodies in venous serum and DBS screen positive if one or more autoantibodies were detected. Participant questionnaires compared the sample collection methods. Of 44 relatives who were multiple autoantibody positive in venous samples, 42 (95.5%) were DBS screen positive, and DBS accurately detected 145 of 147 autoantibody-negative relatives (98.6%). Capillary blood sampling was perceived as more painful than venous blood draw, but 60% of participants would prefer initial screening using home fingerstick with clinic visits only required if autoantibodies were found. Capillary blood sampling could facilitate screening for type 1 diabetes prevention studies.

  20. Nutrition: blood sample collection

    NASA Image and Video Library

    2007-03-20

    ISS014-E-17550 (20 March 2007) --- Astronaut Michael E. Lopez-Alegria, Expedition 14 commander and NASA space station science officer, prepares to insert a test sample in the Minus Eighty Degree Laboratory Freezer for ISS (MELFI) as part of the Nutritional Status Assessment (NUTRITION) experiment in the Destiny laboratory of the International Space Station. MELFI is a low temperature freezer facility with nominal operating temperatures of -80, -26 and +4 degrees Celsius that will preserve experiment materials over long periods.

  1. Nutrition: blood sample collection

    NASA Image and Video Library

    2007-03-20

    ISS014-E-17547 (20 March 2007) --- Astronaut Michael E. Lopez-Alegria, Expedition 14 commander and NASA space station science officer, prepares to insert a test sample in the Minus Eighty Degree Laboratory Freezer for ISS (MELFI) as part of the Nutritional Status Assessment (NUTRITION) experiment in the Destiny laboratory of the International Space Station. MELFI is a low temperature freezer facility with nominal operating temperatures of -80, -26 and +4 degrees Celsius that will preserve experiment materials over long periods.

  2. Noninvasive methods for haemoglobin screening in prospective blood donors.

    PubMed

    Belardinelli, A; Benni, M; Tazzari, P L; Pagliaro, P

    2013-08-01

    The haemoglobin level of prospective blood donors is usually performed on blood obtained by from the finger pulp by fingerstick with a lancet and filling a capillary tube with a sample. New noninvasive methods are now available for rapid, noninvasive predonation haemoglobin screening. Prospective blood donors at our blood centre were tested, in two different trials, as follows: by the NBM 200 (OrSense) test (n = 445 donors) and by the Pronto-7 (Masimo) test (n = 463 donors). The haemoglobin values of each trial and the haemoglobin of finger pulp blood obtained by fingerstick with a lancet (HemoCue) were compared with the haemoglobin values obtained from a venous sample on a Cell Counter (Beckman Coulter). Comparison of Beckman Coulter Cell Counter and OrSense and results showed a bias of 0.29 g/dl, the standard deviation of the differences (SDD) of 0.98 and 95% limits of agreement from -1.64 to 2.21, using Bland and Altman statistical methodology. Comparison of Masimo and Beckman Coulter Cell Counter results showed a bias of -0.53 g/dl, SDD of 1.04 and 95% limits of agreement from -2.57 to 1.51. Cumulative analysis of all 908 donors, as tested by the usual fingerstick test showed a bias of 0.83 g/dl, SDD of 0.70 and 95% limits of agreement from -0.54 to 2.20 compared with the Coulter Cell Counter. Compared with the Coulter Counter, the specificity of the methods was 99.5% for fingerstick, 97% for OrSense and 83% for Massimo, and the sensitivity was 99, 98 and 93%, respectively. Analysis of finger pulp blood by either direct sampling by fingerstick and Hemocue, or by noninvasive haemoglobin tests does not replicate the results of cell counter analysis of venous samples. Compared with fingerstick, noninvasive haemoglobin tests eliminate pain and reduce stress, but have a lower level of specificity and sensitivity. © 2013 International Society of Blood Transfusion.

  3. Population-based screening for anemia using first-time blood donors

    PubMed Central

    Mast, Alan E.; Steele, Whitney R.; Johnson, Bryce; Wright, David J.; Cable, Ritchard G.; Carey, Patricia; Gottschall, Jerome L.; Kiss, Joseph E.; Simon, Toby L.; Murphy, Edward L.

    2012-01-01

    Background Anemia is an important public health concern. Data from population-based surveys such as the National Health and Nutrition Examination Survey (NHANES) are the gold standard, but are obtained infrequently and include only small samples from certain minority groups. Objectives We assessed whether readily available databases of blood donor hemoglobin values could be used as a surrogate for population hemoglobin values from NHANES. Design Blood donor venous and fingerstick hemoglobin values were compared to 10,254 NHANES 2005-2008 venous hemoglobin values using demographically stratified analyses and ANOVA. Fingerstick hemoglobins or hematocrits were converted to venous hemoglobin estimates using regression analysis. Results Venous hemoglobin values from 1,609 first time donors correlated extremely well with NHANES data across different age, gender and demographic groups. Cigarette smoking increased hemoglobin by 0.26 to 0.59 g/dL depending on intensity. Converted fingerstick hemoglobin from 36,793 first time donors agreed well with NHANES hemoglobin (weighted mean hemoglobin of 15.53 g/dL for donors and 15.73 g/dL for NHANES) with similar variation in mean hemoglobin by age. However, compared to NHANES, the larger donor dataset showed reduced differences in mean hemoglobin between Blacks and other races/ethnicities. Conclusions Overall, first-time donor fingerstick hemoglobins approximate U.S. population data and represent a readily available public health resource for ongoing anemia surveillance. PMID:22460662

  4. Factors associated with false-positive results from fingerstick OraQuick ADVANCE rapid HIV 1/2 antibody test.

    PubMed

    Rifkin, Samara B; Owens, Lauren E; Greenwald, Jeffrey L

    2012-01-01

    Identify factors associated with false-positive rapid HIV antibody tests. This retrospective cohort study with nested case-controls involved patients tested for HIV by Boston Medical Center (BMC) affiliates. Cases had a reactive fingerstick OraQuick ADVANCE rapid HIV 1/2 antibody test and a negative Western blot. Controls had nonreactive rapid tests. We compared the prevalence of HIV risk factors between cases and the total nonreactive population and the prevalence of other clinical factors between cases and controls. Of the 15 094 tests, 14 937 (98.9%) were negative and 11 (0.07%) were false positives (specificity of 99.9%). Cases were more likely to have had an HIV-infected sex partner and to be tested at certain sites compared to true negatives. More cases than controls had O-negative blood type. O-negative blood type and sex with an HIV-infected person may increase false-positive HIV fingerstick results. More targeted studies should examine these risk factors.

  5. FDA OKs High-Tech Diabetes Device to Help Replace Fingerstick Tests

    MedlinePlus

    ... Fingerstick Tests Dexcom G5 is first continuous glucose monitoring system that can be used for treatment decisions To ... said Tuesday that the Dexcom G5 continuous glucose monitoring system (CGM) can be used to make insulin dosing ...

  6. Capillary versus venous haemoglobin determination in the assessment of healthy blood donors.

    PubMed

    Patel, A J; Wesley, R; Leitman, S F; Bryant, B J

    2013-05-01

    To determine the accuracy of fingerstick haemoglobin assessment in blood donors, the performance of a portable haemoglobinometer (HemoCue Hb 201+) was prospectively compared with that of an automated haematology analyzer (Cell-Dyn 4000). Haemoglobin values obtained by the latter were used as the 'true' result. Capillary fingerstick samples were assayed by HemoCue in 150 donors. Fingerstick samples from two sites, one on each hand, were obtained from a subset of 50 subjects. Concurrent venous samples were tested using both HemoCue and Cell-Dyn devices. Capillary haemoglobin values (HemoCue) were significantly greater than venous haemoglobin values (HemoCue), which in turn were significantly greater than venous haemoglobin values by Cell-Dyn (mean ± SD: 14.05 ± 1.51, 13.89 ± 1.31, 13.62 ± 1.23, respectively; P < 0.01 for all comparisons among groups). Nine donors (6%) passed haemoglobin screening criteria (≥ 12.5 g/dl) by capillary HemoCue, but were deferred by Cell-Dyn values (false-pass). Five donors (3%) were deferred by capillary sampling, but passed by Cell-Dyn (false-fail). Substantial variability in repeated fingerstick HemoCue results was seen (mean haemoglobin 13.72 vs. 13.70 g/dl, absolute mean difference between paired samples 0.76 g/dl). Hand dominance was not a factor. Capillary samples assessed via a portable device yielded higher haemoglobin values than venous samples assessed on an automated analyzer. False-pass and false-fail rates were low and acceptable in the donor screening setting, with 'true' values not differing by a clinically significant degree from threshold values used to assess acceptability for blood donation. © 2013 The Author(s). Vox Sanguinis © 2013 International Society of Blood Transfusion.

  7. Percutaneous umbilical cord blood sampling - series (image)

    MedlinePlus

    ... your doctor may recommend percutaneous umbilical cord blood sampling (PUBS), which is performed at 18 weeks' gestation. ... it connects to the umbilical cord determine which method your doctor uses. If the placenta is attached ...

  8. Detection of progesterone in whole blood samples.

    PubMed

    Ehrentreich-Förster, Eva; Scheller, Frieder W; Bier, Frank F

    2003-04-01

    The progesterone concentration in blood samples can be utilised as a marker for the diagnosis of early pregnancy, endocrinopathy and virilism. Here, we describe a method for progesterone detection and measurement in whole blood samples by a surface sensitive biosensor used in conjunction with an integrated optical grating coupler. This device determines refractive index changes near the biosensor's surface. Hence, biological species bound to a surface layer can be measured in real-time without any label. For the measurements, we have modified the indirect competitive immunoassay principle. The concentration of the progesterone antibody was kept at 1 microg/ml. Progesterone concentration was determined in buffer solution and whole blood in a range between 0.005 and 10 ng/ml. The detection limit was determined to be 3 pM. The relative standard deviation was calculated to be 3.5%.

  9. Evaluation of home testing to improve follow up after gestational diabetes (Fingerstick Assessments of Sugar Two-months postpartum or FAST)

    PubMed Central

    Lewis, Beth G; Pagan, Elvis R; Evers, Martin

    2013-01-01

    Objective Historically the rates of postpartum glucose tolerance testing for women with gestational diabetes (GDM) average a suboptimal 33%. Barriers include the need for new mothers to miss work and/or arrange for childcare in order to engage in a two-hour test at a commercial lab. This pilot study was initiated to test the theory that a home testing regimen would be accepted by patients and increase the rate of postpartum glucose assessments relative to published rates, without requiring additional health-care staff or resources to achieve this goal. Study design Six weeks postpartum, women with GDM from an academic private practice were asked to check fingerstick blood glucose (FAST Protocol) four times a day for two days, and then obtain an oral glucose tolerance test (OGTT). The physician consultants saw the women each month during pregnancy and arranged the postpartum testing. Results Two of 69 refused to be consented. Twelve of the remaining 67(18%) women completed both the FAST regimen and the OGTT, three completed only the OGTT and five completed only the FAST regimen for a final follow-up rate of 20/67 (30%). The demands of caring for a newborn, or the annoyance of fingersticks, were barriers to compliance. Conclusions In spite of intense physician involvement, this home testing regimen was not associated with an increase in the rates of women participating in postpartum glucose assessments. PMID:27708703

  10. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood...

  11. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood...

  12. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood...

  13. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood...

  14. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood...

  15. Manual versus automated blood sampling: impact of repeated blood sampling on stress parameters and behavior in male NMRI mice.

    PubMed

    Teilmann, A C; Kalliokoski, Otto; Sørensen, Dorte B; Hau, Jann; Abelson, Klas S P

    2014-10-01

    Facial vein (cheek blood) and caudal vein (tail blood) phlebotomy are two commonly used techniques for obtaining blood samples from laboratory mice, while automated blood sampling through a permanent catheter is a relatively new technique in mice. The present study compared physiological parameters, glucocorticoid dynamics as well as the behavior of mice sampled repeatedly for 24 h by cheek blood, tail blood or automated blood sampling from the carotid artery. Mice subjected to cheek blood sampling lost significantly more body weight, had elevated levels of plasma corticosterone, excreted more fecal corticosterone metabolites, and expressed more anxious behavior than did the mice of the other groups. Plasma corticosterone levels of mice subjected to tail blood sampling were also elevated, although less significantly. Mice subjected to automated blood sampling were less affected with regard to the parameters measured, and expressed less anxious behavior. We conclude that repeated blood sampling by automated blood sampling and from the tail vein is less stressful than cheek blood sampling. The choice between automated blood sampling and tail blood sampling should be based on the study requirements, the resources of the laboratory and skills of the staff. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  16. Manual versus automated blood sampling: impact of repeated blood sampling on stress parameters and behavior in male NMRI mice

    PubMed Central

    Kalliokoski, Otto; Sørensen, Dorte B; Hau, Jann; Abelson, Klas S P

    2014-01-01

    Facial vein (cheek blood) and caudal vein (tail blood) phlebotomy are two commonly used techniques for obtaining blood samples from laboratory mice, while automated blood sampling through a permanent catheter is a relatively new technique in mice. The present study compared physiological parameters, glucocorticoid dynamics as well as the behavior of mice sampled repeatedly for 24 h by cheek blood, tail blood or automated blood sampling from the carotid artery. Mice subjected to cheek blood sampling lost significantly more body weight, had elevated levels of plasma corticosterone, excreted more fecal corticosterone metabolites, and expressed more anxious behavior than did the mice of the other groups. Plasma corticosterone levels of mice subjected to tail blood sampling were also elevated, although less significantly. Mice subjected to automated blood sampling were less affected with regard to the parameters measured, and expressed less anxious behavior. We conclude that repeated blood sampling by automated blood sampling and from the tail vein is less stressful than cheek blood sampling. The choice between automated blood sampling and tail blood sampling should be based on the study requirements, the resources of the laboratory and skills of the staff. PMID:24958546

  17. Blood donors and factors impacting the blood donation decision: motives for donating blood in Turkish sample.

    PubMed

    Karacan, Eda; Cengiz Seval, Guldane; Aktan, Zeynep; Ayli, Meltem; Palabiyikoglu, Refia

    2013-12-01

    Donations in Turkey are insufficient to cover the high transfusion needs arising from large numbers of thalassemia and sickle cell anemia patients and increasing demands for blood due to advanced surgery and cancer treatment. The most acceptable means to get blood is voluntary blood donation and the blood donor system in Turkey mostly depends on a combination of voluntary and involuntary donors. The main aim of this study is to explore the motivations of Turkish voluntary blood donors toward blood donation and to determine predictors of blood donation motivation. A cross-sectional sample survey of active blood donors in Ankara, Turkey was conducted. The sample consisted of 189 male volunteer blood donor adults. Donors filled in a self-administered questionnaire including the measures of demographic information, empathetic concern, altruism, social responsibility and blood donation motivation questionnaire during donation. Factor analysis of Blood Donation Motivation Measure with varimax rotation revealed a three-factor solution named as "values and moral duty", "positive feelings and esteem" and "self-benefit and external reasons". The results with regression analyses showed that only social responsibility had an significant effect independent of age, income, and education on blood donation motivation. These result reflects that blood donation motivation not only linked to a high degree of altruistic reasons, but also to a combination of some self-regarding motives. Additionally, feelings of empathy or altruism may be less strong at the time the decision to help, other factors may have a larger influence on helping decisions. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. Factors affecting blood sample haemolysis: a cross-sectional study.

    PubMed

    Barnard, Ed B G; Potter, David L; Ayling, Ruth M; Higginson, Ian; Bailey, Andrew G; Smith, Jason E

    2016-04-01

    To determine the effect of blood sampling through an intravenous catheter compared with a needle in Emergency Department blood sampling. We undertook a prospective, cross-sectional study in a UK university teaching hospital Emergency Department. A convenience sample of 985 patients who required blood sampling via venepuncture was collected. A total of 844 complete sets of data were analysed. The median age was 63 years, and 57% of patients were male. The primary outcome measure was the incidence of haemolysis in blood samples obtained via a needle compared with samples obtained via an intravenous catheter. Secondary outcome measures defined the effect on sample haemolysis of the side of the patient the sample was obtained from, the anatomical location of sampling, the perceived difficulty in obtaining the sample, the order of sample tubes collected, estimated tourniquet time and bench time. Data were analysed with logistic regression, and expressed as odds ratios (95% confidence intervals; P-values). Blood samples obtained through an intravenous catheter were more likely to be haemolysed than those obtained via a needle, odds ratio 5.63 (95% confidence interval 2.49-12.73; P<0.001). Blood sampling via an intravenous catheter was significantly associated with an increase in the likelihood of sample haemolysis compared with sampling with a needle. Wherever practicable, blood samples should be obtained via a needle in preference to an intravenous catheter. Future research should include both an economic evaluation, and staff and patient satisfaction of separating blood sampling and intravenous catheter placement.

  19. Guide to capillary heelstick blood sampling in infants.

    PubMed

    Folk, Laura A

    2007-08-01

    Capillary blood sampling is an essential method of blood collection performed by nurses of all skill levels to obtain samples for routine laboratory tests in neonates. Accuracy of results depends on proper heelstick and sample collection technique. Recent advances including development of devices designed specifically for heelstick capillary blood sampling and research into expanded safe heel capillary sampling sites are discussed. A step-by-step guide to capillary blood sampling is outlined along with evidence-based practice incorporating neonatal-appropriate disinfection and nonpharmacological analgesia that contribute to improved infant safety and comfort during and after the procedure.

  20. Hemolysis associated with pneumatic tube system transport for blood samples.

    PubMed

    Kara, Hasan; Bayir, Aysegul; Ak, Ahmet; Degirmenci, Selim; Akinci, Murat; Agacayak, Ahmet; Marcil, Emine; Azap, Melih

    2014-01-01

    The frequency of hemolysis of blood samples may be increased by transport in a pneumatic tube system. The purpose of this study was to evaluate the effect of pneumatic tube system transport on hemolysis of blood samples. Blood samples were transported from the emergency department to the hospital laboratory manually by hospital staff (49 patients) or with a pneumatic tube system (53 patients). The hemolysis index and serum chemistry studies were performed on the blood samples and compared between the different methods of transport. The blood samples that were transported by the pneumatic tube system had a greater frequency of hemolysis and greater mean serum potassium and median creatinine, aspartate aminotransferase, and lactate dehydrogenase levels than samples transported manually. Blood samples transported from the emergency department to the hospital laboratory by a pneumatic tube system may have a greater frequency of hemolysis than samples transported manually. This may necessitate repeat phlebotomy and cause a delay in completing the laboratory analysis.

  1. Combination syringe provides air-free blood samples

    NASA Technical Reports Server (NTRS)

    Pool, S. L.

    1970-01-01

    Standard syringe and spinal needle are combined in unique manner to secure air-free blood samples. Combination syringe obtains air free samples because air bubbles become insignificant when samples greater than 1 cc are drawn.

  2. The Potential for Glycemic Control Monitoring and Screening for Diabetes at Dental Visits Using Oral Blood

    PubMed Central

    Rosedale, Mary T.; Pesce, Michael A.; Rindskopf, David M.; Kaur, Navjot; Juterbock, Caroline M.; Wolff, Mark S.; Malaspina, Dolores; Danoff, Ann

    2015-01-01

    Objectives. We examined the potential for glycemic control monitoring and screening for diabetes in a dental setting among adults (n = 408) with or at risk for diabetes. Methods. In 2013 and 2014, we performed hemoglobin A1c (HbA1c) tests on dried blood samples of gingival crevicular blood and compared these with paired “gold-standard” HbA1c tests with dried finger-stick blood samples in New York City dental clinic patients. We examined differences in sociodemographics and diabetes-related risk and health care characteristics for 3 groups of at-risk patients. Results. About half of the study sample had elevated HbA1c values in the combined prediabetes and diabetes ranges, with approximately one fourth of those in the diabetes range. With a correlation of 0.991 between gingival crevicular and finger-stick blood HbA1c, measures of concurrence between the tests were extremely high for both elevated HbA1c and diabetes-range HbA1c levels. Persons already diagnosed with diabetes and undiagnosed persons aged 45 years or older could especially benefit from HbA1c testing at dental visits. Conclusions. Gingival crevicular blood collected at the dental visit can be used to screen for diabetes and monitor glycemic control for many at-risk patients. PMID:25713975

  3. Whole Genome Amplification from Blood Spot Samples.

    PubMed

    Sørensen, Karina Meden

    2015-01-01

    Whole genome amplification is an invaluable technique when working with DNA extracted from blood spots, as the DNA obtained from this source often is too limited for extensive genetic analysis. Two techniques that amplify the entire genome are common. Here, both are described with focus on the benefits and drawbacks of each system. However, in order to obtain the best possible WGA result the quality of input DNA extracted from the blood spot is essential, but also time consumption, flexibility in format and elution volume and price of the technology are factors influencing system choice. Here, three DNA extraction techniques are described and the above aspects are compared between the systems.

  4. A technique for improved blood sampling during sleep studies.

    PubMed

    Ona, E; Dimsdale, J E; Ancoli-Israel, S; Dillon, E; Watkins, L; Coy, T V; Clausen, J

    1994-11-01

    Research protocols often require that blood samples be drawn during sleep. This study compares the efficacy of obtaining nocturnal blood samples using a standard heparinized intravenous setup versus the same intravenous setup used in conjunction with a small chemical heating pad. The chemical heating pad significantly improved the number of blood samples obtained and the maintenance of intravenous patency. The use of a chemical heating pad is an economical way to resolve the frustration of lost blood samples while maintaining a reasonable environment to monitor sleep.

  5. Payload specialist Reinhard Furrer show evidence of previous blood sampling

    NASA Technical Reports Server (NTRS)

    1985-01-01

    Payload specialist Reinhard Furrer shows evidence of previous blood sampling while Wubbo J. Ockels, Dutch payload specialist (only partially visible), extends his right arm after a sample has been taken. Both men show bruises on their arms.

  6. Blood oxygen saturation determined by transmission spectrophotometry of hemolyzed blood samples

    NASA Technical Reports Server (NTRS)

    Malik, W. M.

    1967-01-01

    Use of the Lambert-Beer Transmission Law determines blood oxygen saturation of hemolyzed blood samples. This simplified method is based on the difference in optical absorption properties of hemoglobin and oxyhemoglobin.

  7. Sampling problems in the micro determination of blood lead.

    PubMed

    Juselius, R E; Lupovich, P; Moriarty, R

    1975-01-01

    Sampling tube and fingertip contamination were found to present potential problems in the collection of samples for micro blood lead analyses. Large differences between micro screening and macro confirming lead levels were frequently observed when the time between collection of the two samples was 1-2 weeks. The magnitude of these differences decreased as macro blood lead concentration increased and were apparently a result of episodic lead ingestion in the population.

  8. Hemolysis associated with pneumatic tube system transport for blood samples

    PubMed Central

    Kara, Hasan; Bayir, Aysegul; Ak, Ahmet; Degirmenci, Selim; Akinci, Murat; Agacayak, Ahmet; Marcil, Emine; Azap, Melih

    2014-01-01

    Objective: The frequency of hemolysis of blood samples may be increased by transport in a pneumatic tube system. The purpose of this study was to evaluate the effect of pneumatic tube system transport on hemolysis of blood samples. Methods: Blood samples were transported from the emergency department to the hospital laboratory manually by hospital staff (49 patients) or with a pneumatic tube system (53 patients). The hemolysis index and serum chemistry studies were performed on the blood samples and compared between the different methods of transport. Results: The blood samples that were transported by the pneumatic tube system had a greater frequency of hemolysis and greater mean serum potassium and median creatinine, aspartate aminotransferase, and lactate dehydrogenase levels than samples transported manually. Conclusion: Blood samples transported from the emergency department to the hospital laboratory by a pneumatic tube system may have a greater frequency of hemolysis than samples transported manually. This may necessitate repeat phlebotomy and cause a delay in completing the laboratory analysis. PMID:24639830

  9. Assessment Of Preanalytical Blood Sampling Errors In Clinical Settings.

    PubMed

    Zehra, Nayyab; Malik, Ahmed Hassaan; Arshad, Qurat; Sarwar, Sumaira; Aslam, Sehar

    2016-01-01

    Blood sampling is one of the common procedures done in every ward for disease diagnosis and prognosis. Daily hundreds of samples are collected from different wards but lack of appropriate knowledge of blood sampling by paramedical staff and accidental errors make the samples inappropriate for testing. Thus the need to avoid these errors for better results still remains. We carried out this research with an aim to determine the common errors during blood sampling; find factors responsible and propose ways to reduce these errors. A cross sectional descriptive study was carried out at the Military and Combined Military Hospital Rawalpindi during February and March 2014. A Venous Blood Sampling questionnaire (VBSQ) was filled by the staff on voluntary basis in front of the researchers. The staff was briefed on the purpose of the survey before filling the questionnaire. Sample size was 228. Results were analysed using SPSS-21. When asked in the questionnaire, around 61.6% of the paramedical staff stated that they cleaned the vein by moving the alcohol swab from inward to outwards while 20.8% of the staff reported that they felt the vein after disinfection. On contrary to WHO guidelines, 89.6% identified that they had a habit of placing blood in the test tube by holding it in the other hand, which should actually be done after inserting it into the stand. Although 86% thought that they had ample knowledge regarding the blood sampling process but they didn't practice it properly. Pre analytical blood sampling errors are common in our setup. Eighty six percent participants though thought that they had adequate knowledge regarding blood sampling, but most of them were not adhering to standard protocols. There is a need of continued education and refresher courses.

  10. Are They Bloody Guilty? Blood Doping with Simulated Samples

    ERIC Educational Resources Information Center

    Stuart, Parker E.; Lees, Kelsey D.; Milanick, Mark A.

    2014-01-01

    In this practice-based lab, students are provided with four Olympic athlete profiles and simulated blood and urine samples to test for illegal substances and blood-doping practices. Throughout the course of the lab, students design and conduct a testing procedure and use their results to determine which athletes won their medals fairly. All of the…

  11. Are They Bloody Guilty? Blood Doping with Simulated Samples

    ERIC Educational Resources Information Center

    Stuart, Parker E.; Lees, Kelsey D.; Milanick, Mark A.

    2014-01-01

    In this practice-based lab, students are provided with four Olympic athlete profiles and simulated blood and urine samples to test for illegal substances and blood-doping practices. Throughout the course of the lab, students design and conduct a testing procedure and use their results to determine which athletes won their medals fairly. All of the…

  12. Non-terminal blood sampling techniques in guinea pigs.

    PubMed

    Birck, Malene M; Tveden-Nyborg, Pernille; Lindblad, Maiken M; Lykkesfeldt, Jens

    2014-10-11

    Guinea pigs possess several biological similarities to humans and are validated experimental animal models(1-3). However, the use of guinea pigs currently represents a relatively narrow area of research and descriptive data on specific methodology is correspondingly scarce. The anatomical features of guinea pigs are slightly different from other rodent models, hence modulation of sampling techniques to accommodate for species-specific differences, e.g., compared to mice and rats, are necessary to obtain sufficient and high quality samples. As both long and short term in vivo studies often require repeated blood sampling the choice of technique should be well considered in order to reduce stress and discomfort in the animals but also to ensure survival as well as compliance with requirements of sample size and accessibility. Venous blood samples can be obtained at a number of sites in guinea pigs e.g., the saphenous and jugular veins, each technique containing both advantages and disadvantages(4,5). Here, we present four different blood sampling techniques for either conscious or anaesthetized guinea pigs. The procedures are all non-terminal procedures provided that sample volumes and number of samples do not exceed guidelines for blood collection in laboratory animals(6). All the described methods have been thoroughly tested and applied for repeated in vivo blood sampling in studies within our research facility.

  13. Clinical evaluation of a novel on-strip calibration method for blood glucose measurement.

    PubMed

    Noble, Michael; Rippeth, John; Edington, David; Rayman, Gerry; Brandon-Jones, Sarah; Hollowood, Zoe; Kew, Simon

    2014-07-01

    This study evaluated a novel technology for improving accuracy of self-monitoring of blood glucose (SMBG). The technology calibrates each and every test by measuring the response from a predetermined amount of glucose present in the sample chamber of each test strip. SMBG test strips were modified to include a lid coated with a fast dissolving formulation containing glucose. These test strips were characterized for hematocrit (Hct) and temperature induced error response to develop a calibration algorithm. The modified test strips were used in a clinical evaluation involving fingerstick blood samples from 160 subjects. Experiments involving Hct and temperature induced errors show that the technology generates a signal characteristic of the error conditions in any particular test, but independent of glucose concentration, allowing a correction algorithm to be derived. The approach substantially reduced Hct and temperature derived errors. Clinical evaluation using fingerstick blood directly applied to prototype strips showed the error (measured as MARD) was reduced from 11.1 to 5.9% by the on-strip correction approach and the number of outliers reduced by approximately 90%. This technology could improve the accuracy and precision of glucose monitoring systems and so reduce decision errors particularly in clinical situations where hematocrit and temperature may be significant confounders.

  14. Astronaut Joseph Kerwin takes blood sample from Astronaut Charles Conrad

    NASA Technical Reports Server (NTRS)

    1973-01-01

    Scientist-Astronaut Joseph P. Kerwin (right), Skylab 2 science pilot and a doctor of medicine, takes a blood sample from Astronaut Charles Conrad Jr., Sylab 2 commander, as seen in this reproduction taken from a color television transmission made by a TV camera aboard the Skylab 1 and 2 space station cluster in Earth orbit. The blood sampling was part of the Skylab Hematology and Immunology Experiment M110 series.

  15. Astronaut Joseph Kerwin takes blood sample from Astronaut Charles Conrad

    NASA Technical Reports Server (NTRS)

    1973-01-01

    Scientist-Astronaut Joseph P. Kerwin (right), Skylab 2 science pilot and a doctor of medicine, takes a blood sample from Astronaut Charles Conrad Jr., Sylab 2 commander, as seen in this reproduction taken from a color television transmission made by a TV camera aboard the Skylab 1 and 2 space station cluster in Earth orbit. The blood sampling was part of the Skylab Hematology and Immunology Experiment M110 series.

  16. Coagulation dynamics of a blood sample by multiple scattering analysis

    NASA Astrophysics Data System (ADS)

    Faivre, Magalie; Peltié, Philippe; Planat-Chrétien, Anne; Cosnier, Marie-Line; Cubizolles, Myriam; Nougier, Christophe; Négrier, Claude; Pouteau, Patrick

    2011-05-01

    We report a new technique to measure coagulation dynamics on whole-blood samples. The method relies on the analysis of the speckle figure resulting from a whole-blood sample mixed with coagulation reagent and introduced in a thin chamber illuminated with a coherent light. A dynamic study of the speckle reveals a typical behavior due to coagulation. We compare our measured coagulation times to a reference method obtained in a medical laboratory.

  17. Evaluating venous pool technique for blood sampling in neonatal ICU.

    PubMed

    Hatler, Carol; Dalton, Beverly; Day, Susan; Sharfner, Andrea; Hauffe, Rhonda

    2013-01-01

    To evaluate venous pool technique (VPT) for obtaining neonatal blood samples as compared with the needlestick technique. An experimental design was used with subjects enrolled in two phases: an equivalence phase (N = 10) and a comparison phase (N = 64). In the equivalence phase, subjects weighing 1,500 g or more had two needlesticks. In the comparison phase, subjects weighing 800 g or more were randomized to receive blood drawn by either needlestick method or VPT. Comparative results suggest that infant and maternal demographic factors, sampling attempts, and sampling failures were similar. However, for the outcome of hematoma development, the standard technique was significantly worse (t = 2.25 ; p = .029). Results suggest that the VPT method is safe and accurate for use in critically ill neonates. This study demonstrated that the VPT process is easily learned and may provide advantages over standard blood sampling methods. Nurses can use this information to evaluate this VPT technique in their institutions.

  18. [Blood sampling using "dried blood spot": a clinical biology revolution underway?].

    PubMed

    Hirtz, Christophe; Lehmann, Sylvain

    2015-01-01

    Blood testing using the dried blood spot (DBS) is used since the 1960s in clinical analysis, mainly within the framework of the neonatal screening (Guthrie test). Since then numerous analytes such as nucleic acids, small molecules or lipids, were successfully measured on the DBS. While this pre-analytical method represents an interesting alternative to classic blood sampling, its use in routine is still limited. We review here the different clinical applications of the blood sampling on DBS and estimate its future place, supported by the new methods of analysis as the LC-MS mass spectrometry.

  19. Measurement and Comparison of Organic Compound Concentrations in Plasma, Whole Blood, and Dried Blood Spot Samples.

    PubMed

    Batterman, Stuart A; Chernyak, Sergey; Su, Feng-Chiao

    2016-01-01

    The preferred sampling medium for measuring human exposures of persistent organic compounds (POPs) is blood, and relevant sample types include whole blood, plasma, and dried blood spots (DBS). Because information regarding the performance and comparability of measurements across these sample types is limited, it is difficult to compare across studies. This study evaluates the performance of POP measurements in plasma, whole blood and DBS, and presents the distribution coefficients needed to convert concentrations among the three sample types. Blood samples were collected from adult volunteers, along with demographic and smoking information, and analyzed by GC/MS for organochlorine pesticides (OCPs), chlorinated hydrocarbons (CHCs), polychlorinated biphenyls (PCBs), and brominated diphenyl ethers (PBDEs). Regression models were used to evaluate the relationships between the sample types and possible effects of personal covariates. Distribution coefficients also were calculated using physically-based models. Across all compounds, concentrations in plasma were consistently the highest; concentrations in whole blood and DBS samples were comparable. Distribution coefficients for plasma to whole blood concentrations ranged from 1.74 to 2.26 for pesticides/CHCs, averaged 1.69 ± 0.06 for the PCBs, and averaged 1.65 ± 0.03 for the PBDEs. Regression models closely fit most chemicals (R (2) > 0.80), and whole blood and DBS samples generally showed very good agreement. Distribution coefficients estimated using biologically-based models were near one and did not explain the observed distribution. Among the study population, median concentrations of several pesticides/CHCs and PBDEs exceeded levels reported in the 2007-2008 National Health and Nutrition Examination Survey, while levels of other OCPs and PBDEs were comparable or lower. Race and smoking status appeared to slightly affect plasma/blood concentration ratios for several POPs. The experimentally

  20. Determination of chlorinated insecticides in blood samples of agricultural workers.

    PubMed

    Rosell, M G; Obiols, J; Berenguer, M J; Guardino, X; López, F; Brosa, J

    1993-11-26

    Lindane, aldrin and p,p'-DDT were determined in blood samples from 71 farmers by means of an analytical method which combines a direct whole-blood extraction with n-hexane and gas chromatography (GC)-electron-capture detection (ECD), using a capillary column, applied to the organic extract. This technique allowed the determination of pesticides at levels varying from 0.1 to 180 micrograms per l of blood, the detection limit for every pesticide being 0.1 microgram/l. GC-mass spectrometry was used to confirm the identity of each pesticide. The advantage of capillary column GC-ECD for pesticide determination is its sensitivity and high resolution, which makes it possible to separate pesticides from a complex n-hexane extract obtained in a very simple pretreatment of the blood sample, which is itself a very complex matrix.

  1. Is supine rest necessary before blood sampling for plasma metanephrines?

    PubMed

    Lenders, Jacques W M; Willemsen, Jacques J; Eisenhofer, Graeme; Ross, H Alec; Pacak, Karel; Timmers, Henri J L M; Sweep, C G J Fred

    2007-02-01

    The impact of blood sampling in sitting vs supine positions on measurements of plasma metanephrines for diagnosis of pheochromocytoma is unknown. We compared plasma concentrations of free metanephrines in samples from patients with primary hypertension obtained after supine rest with those obtained in the sitting position without preceding rest. We also assessed the effects on diagnostic test performance retrospectively in patients with and without pheochromocytoma, and we calculated cost-effectiveness for pheochromocytoma testing. Upper reference limits of plasma free metanephrines were higher in samples obtained from seated patients without preceding rest than from supine patients with preceding rest. Application of these higher upper reference limits to samples from supine patients with pheochromocytoma decreased the diagnostic sensitivity from 99% to 96%. In patients without pheochromocytoma, adjusting the plasma concentration for the effects of sitting while preserving the 99% sensitivity by use of the supine upper reference limits increased the number of false-positive test results from 9% to 25%. To preserve high diagnostic sensitivity we recommend the use of upper reference limits determined from blood samples collected in the supine position. Under these conditions, negative test results for blood samples obtained with patients sitting are as effective for ruling out pheochromocytoma as negative results from samples obtained after supine rest. Repeat testing with samples obtained in the supine position offers a cost-effective approach for dealing with the increased numbers of false-positive results expected after initial sampling in the sitting position.

  2. Determination of blood cell subtype concentrations from frozen whole blood samples using TruCount beads.

    PubMed

    Langenskiöld, Cecilia; Mellgren, Karin; Abrahamsson, Jonas; Bemark, Mats

    2016-06-24

    In many studies it would be advantageous if blood samples could be collected and analyzed using flow cytometry at a later stage. Ideally, sample collection should involve little hands-on time, allow for long-term storage, and minimally influence the samples. Here we establish a flow cytometry antibody panel that can be used to determine granulocytes, monocytes, and lymphocyte subset concentrations in fresh and frozen whole blood using TruCount technology. The panel can be used on fresh whole-blood samples as well as whole-blood samples that have been frozen after mixing with 10% DMSO. Concentrations in frozen and fresh sample is highly correlated both when frozen within 4 h and the day after collection (r ≥ 0.98), and the estimated concentration in frozen samples was between 91 and 94% of that in fresh samples for all cell types. Using this method whole-blood samples can be frozen using a simple preparation method, and stored long-term before accurate determination of cell concentration. This allows for standardized analysis of the samples at a reference laboratory in multi-center studies. © 2016 International Clinical Cytometry Society. © 2016 International Clinical Cytometry Society.

  3. Viscosity measurements on very small capillary blood samples.

    PubMed

    Eugster, M; Häusler, K; Reinhart, W H

    2007-01-01

    Viscosity measurements on very small capillary blood samples could be of considerable clinical interest. We have developed an oscillating viscometer for very small volumes, which consists of a glass capillary containing 7 mul of blood, which is part of an oscillating torsional resonator. The damping of the sinusoidal oscillations depends on the density and viscosity of the fluid, which allows blood viscosity measurements. The instrument was first evaluated in comparison with a standard blood viscometer (Contraves LS 30). Blood from healthy volunteers anticoagulated with EDTA was adjusted to hematocrit levels of 20, 30, 40, 50, and 60%, respectively. A strong correlation was found between hematocrit and oscillating viscosity (y=0.17x-2.05, r=0.969, p<0.0001) and between oscillating and conventional high shear viscosity (y=1.11x-0.62, r=0.971, p<0.0001). Blood viscosity measured in venous or capillary blood of normal subjects was similar (p=0.63). Bedside viscosity measurements on capillary blood drawn from a finger prick during routine blood glucose measurements in patients with diabetes mellitus showed lower blood viscosity than controls (3.62+/-0.87 vs 4.79+/-0.59 mPa.s, p=0.0007), which is in contrast to earlier publications, and may be explained by the lower hematocrit in our diabetic patients (34.7+/-6.0% vs. 43.1+/-1.9%, p<0.0001). Blood viscosity was independent of the actual glucose level (range 3-17 mmol/l). Capillary blood anticoagulated with EDTA was drawn by heel prick from 23 newborns. Blood viscosity was higher (5.66 +/-2.47 mPa.s) than in adult controls (see above), which could be explained by the dependence on the higher hematocrit (46.4 +/-8.6%). We conclude that viscosity measurements can be made on very small samples such as capillary blood from diabetic patients or newborn babies with this new oscillating viscometer. It remains to be determined if such new informations have clinical implications.

  4. Polybrominated diphenyl ethers in maternal and fetal blood samples.

    PubMed Central

    Mazdai, Anita; Dodder, Nathan G; Abernathy, Mary Pell; Hites, Ronald A; Bigsby, Robert M

    2003-01-01

    Polybrominated diphenyl ethers (PBDEs) are widely used as flame retardants in consumer goods, such as plastics, electronics, textiles, and construction material. PBDEs have been found in human milk, fat, and blood samples. Rodent studies indicate that PBDEs may be detrimental to neurodevelopment, possibly by lowering thyroid hormone concentrations in blood. In the present study, we determined concentrations of PBDEs and thyroid hormones in human fetal and maternal serum. Patients presenting in labor to Indiana University and Wishard Memorial County hospitals in Indianapolis, who were older than 18 years, were recruited to participate. Twelve paired samples of maternal and cord blood were obtained and analyzed using gas chromatographic mass spectrometry; thyroid hormone concentrations were determined by radioimmunoassay. Six congeners of PBDE were measured in maternal and fetal serum samples. The concentrations of total PBDEs found in maternal sera ranged from 15 to 580 ng/g lipid, and the concentrations found in fetal samples ranged from 14 to 460 ng/g lipid. Individual fetal blood concentrations did not differ from the corresponding maternal concentrations, indicating that measurement of maternal PBDE blood levels is useful in predicting fetal exposure; similarly, other reports have shown a high correlation between PBDE in mother's milk and fetal exposure. In accord with reports on other biologic samples, the tetrabrominated PBDE congener BDE-47 accounted for 53-64% of total PBDEs in the serum. The concentrations of PBDEs found in maternal and fetal serum samples were 20-106-fold higher than the levels reported previously in a similar population of Swedish mothers and infants. In this small sample, there was no apparent correlation between serum PBDEs and thyroid hormone concentrations. Our study shows that human fetuses in the United States may be exposed to relatively high levels of PBDEs. Further investigation is required to determine if these levels are

  5. Quality of plasma sampled by different methods for multiple blood sampling in mice.

    PubMed

    Christensen, S D; Mikkelsen, L F; Fels, J J; Bodvarsdóttir, T B; Hansen, A K

    2009-01-01

    For oral glucose tolerance test (OGTT) in mice, multiple blood samples need to be taken within a few hours from conscious mice. Today, a number of essential parameters may be analysed on very small amounts of plasma, thus reducing the number of animals to be used. It is, however, crucial to obtain high-quality plasma or serum in order to avoid increased data variation and thereby increased group sizes. The aim of this study was to find the most valid and reproducible method for withdrawal of blood samples when performing OGTT. Four methods, i.e. amputation of the tail tip, lateral tail incision, puncture of the tail tip and periorbital puncture, were selected for testing at 21 degrees C and 30 degrees C after a pilot study. For each method, four blood samples were drawn from C57BL/6 mice at 30 min intervals. The presence of clots was registered, haemolysis was monitored spectrophotometrically at 430 nm, and it was noted whether it was possible to achieve 30-50 microL blood. Furthermore, a small amount of extra blood was sampled before and after the four samplings for testing of whether the sampling induced a blood glucose change over the 90 min test period. All methods resulted in acceptable amounts of plasma. Clots were observed in a sparse number of samples with no significant differences between the methods. Periorbital puncture did not lead to any haemolysed samples at all, and lateral tail incision resulted in only a few haemolysed samples, while puncture or amputation of the tail tip induced haemolysis in a significant number of samples. All methods, except for puncture of the tail tip, influenced blood glucose. Periorbital puncture resulted in a dramatic increase in blood glucose of up to 3.5 mmol/L indicating that it is stressful. Although lateral tail incision also had some impact on blood glucose, it seems to be the method of choice for OGTT, as it is likely to produce a clot-free non-haemolysed sample, while periorbital sampling, although producing a

  6. Effects of post-sampling analysis time, type of blood samples and collection tubes on values of blood gas testing.

    PubMed

    Smajić, Jasmina; Kadić, Damira; Hasić, Sabaheta; Serdarević, Nafija

    2015-08-01

    To investigate effects of post-sampling analysis time, a type of blood samples and collection tubes on blood gas testing. This study included 100 patients at the Clinic for Pulmonary Diseases, Clinical Centre Sarajevo. The partial pressure of oxygen (pO2) and carbon dioxide (pCO2), and the oxygen saturation level of hemoglobin (sO2) were analyzed in the arterial blood samples (ABS) and capillary blood samples (CBS) by a potentiometric method using a blood gas analyzer ABL 555 (Radiometer, Copenhagen, Denmark). Paired measurements of ABS were performed within 15 minutes and after 60 minutes from sampling and compared. The results of CBS obtained within 15 minutes were compared with matching ABS results, as well as the results obtained from CBS within 15 minutes taken into glass and plastic tubes. pO2 and sO2 values were significantly lower after 60 minutes compared to those within 15 minutes in ABS (9.20±1.89 vs. 9.51±1.95 and 91.25±5.03 vs. 92.40±4.5; p<0.01, respectively). Values of pO2 and sO2 in CBS were significantly lower than values obtained in ABS (8.92±2.07 vs. 9.51±1.95 and 91.25±4.86 vs. 92.40±4.50; p<0.01, respectively). Obtained pO2 and sO2 values in CBS in the plastic tubes were higher than those in the glass tubes (8.50±1.98 vs. 7.89±2.0 and 89.66±11.04 vs. 88.23±11.22, p<0.01 respectively). pCO2 blood values were not influenced significantly (p>0.05). The length of post-sampling analysis time, a type of blood samples and collection tubes have significant impact on blood oxygen parameters. Analysis within 15 minutes after blood sampling is considered as appropriate. Copyright© by the Medical Assotiation of Zenica-Doboj Canton.

  7. Improved sample capsule for determination of oxygen in hemolyzed blood

    NASA Technical Reports Server (NTRS)

    Malik, W. M.

    1967-01-01

    Sample capsule for determination of oxygen in hemolyzed blood consists of a measured section of polytetrafluoroethylene tubing equipped at each end with a connector and a stopcock valve. This method eliminates errors from air entrainment or from the use of mercury or syringe lubricant.

  8. Bedside capillary blood glucose measurements in critically ill patients: influence of catecholamine therapy.

    PubMed

    Fekih Hassen, M; Ayed, S; Gharbi, R; Ben Sik Ali, H; Marghli, S; Elatrous, S

    2010-01-01

    To evaluate the effects of catecholamine therapy on the accuracy of capillary glucose measurements in hyperglycemic patients. 43 hyperglycemic patients older than 18 years admitted to the intensive care unit of a tertiary medical center were included from December 2005 to March 2006. This prospective study compares fingerstick and earlobe measurements simultaneously to sampled laboratory venous glucose in patients treated without (group 1) or with (group 2) catecholamine. Three venous samples for serum glucose analysis at three fixed hours and simultaneously two capillary glucose determinations were performed during the two first successive days after inclusion. A difference between the methods of glucose measurements greater than 2.3 mmol/l was considered significant. The mean difference between the two methods was -0.05 mmol/l in group 1 and +0.29 mmol/l in group 2, while the limits of agreement were +4.03 and -4.13 mmol/l and +5.63 and -5.05 mmol/l in groups 1 and 2, respectively. A difference between paired measurements greater than 2.3 mmol/l was observed in 29% in group 1 and in 40% in group 2 (p=0.038). The alternative site did not improve the accuracy. In critically ill patients treated with catecholamine capillary fingerstick blood glucose measurement seems inaccurate. Earlobe sampling does not improve accuracy. Copyright 2009 Elsevier Ireland Ltd. All rights reserved.

  9. Design of Er:YAG laser blood-sampling device

    NASA Astrophysics Data System (ADS)

    Wu, Zhi-chao; Jin, Guang-yong; Tan, Xue-chun; Ling, Ming; Liang, Zhu

    2009-07-01

    Laser blood-sampling device is one of the foremost tasks in medicine domain. It has a lot of merits such as un-touching, avoiding infection, indolence, and fast healing etc. The Er:YAG laser with wavelength of 2.94μm which is just close to the absorbency peak of water can be strongly absorbed by water molecular, so it has very wide application value in clinical medicine. In the paper, based on the mutual action characters of the laser with 2.94μm wave length on biological tissues, such as high absorption, acting on surface, the design of a new type of laser blood-sampling device is introduced. According to the needs of practice, the main component of the blood-sampling device is the laser, which includes optical resonator, optical collector, pumping source, optical guidance and focusing system. All of them are designed in the paper, and the reflection index of output coupling mirror of laser is optimized, the laser threshold is reduced, and pumping efficiency is improved. Moreover, thermal effect of Er:YAG solid-state laser is analyzed and a reasonable cooling method is designed. As a result, an excellent laser blood- sampling is obtained, the maximum output power is about 1J, the optical to optical conversion efficiency is 1.2%. For the better production-grade, the cuprum-based conduction is adopt to eliminate heat, the precision modulation and fixing of the optical resonance is achieved by the special adjusting structure that not only improve the stability and reliability, but also reduce the size of laser bloodsampling device. The size is 110×190×320mm, the weight is about 5.8kg, and the laser blood- sampling efficiency is 100%.

  10. Quantification of multiple elements in dried blood spot samples.

    PubMed

    Pedersen, Lise; Andersen-Ranberg, Karen; Hollergaard, Mads; Nybo, Mads

    2017-08-01

    Dried blood spots (DBS) is a unique matrix that offers advantages compared to conventional blood collection making it increasingly popular in large population studies. We here describe development and validation of a method to determine multiple elements in DBS. Elements were extracted from punches and analyzed using inductively coupled plasma-mass spectrometry (ICP-MS). The method was evaluated with quality controls with defined element concentration and blood spiked with elements to assess accuracy and imprecision. DBS element concentrations were compared with concentrations in venous blood. Samples with different hematocrit were spotted onto filter paper to assess hematocrit effect. The established method was precise and accurate for measurement of most elements in DBS. There was a significant but relatively weak correlation between measurement of the elements Mg, K, Fe, Cu, Zn, As and Se in DBS and venous whole blood. Hematocrit influenced the DBS element measurement, especially for K, Fe and Zn. Trace elements can be measured with high accuracy and low imprecision in DBS, but contribution of signal from the filter paper influences measurement of some elements present at low concentrations. Simultaneous measurement of K and Fe in DBS extracts may be used to estimate sample hematocrit. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  11. [Factors related to haemolysis in the extraction of blood samples].

    PubMed

    Agós, M D; Lizarraga, R; Gambra, D; Marañón, A; Orozco, C; Díaz, E

    2008-01-01

    The choice of a venous access system to provide safe blood collection and reliable analytical results for that sample is of paramount importance in any accident and emergency department. The objective of this study was to identify the factors associated with haemolysis in venous blood samples, where the variables studied were: type of venipuncture (needle and catheter), type of catheter (3 catheters of 3 different materials) and diameter of the catheter. The sample was obtained from all patients who required a blood test in the accident and emergencies department of the Virgen del Camino Hospital over 34 days, collected in 3 different periods (September-November), involving a total of 1.933 procedures. Positive haemolysis determined by laboratory technicians was found in 2% (7/348) of samples obtained by needle compared to 14% (222/1585) obtained by catheter. We observe an 8% (39/475) of haemolysis in the samples taken by protective Teflon catheter, 18% (77/426) by Protectiv plus polyurethane and 15% (106/684) by BD-Nexiva Vialone. The haemolysis index fell with an increase in the size of the catheter, those of 18G showing 13% (115/867) and those of 20G showing 15% (107/708). The combination of catheter type and size maintains the smallest percentages of haemolysis in Teflon catheters and high diameters of 18G with 6% (19/301), less than half the haemolysis of the polyurethane catheters and a third of that for Vialone catheters respectively.

  12. Therapy monitoring in congenital adrenal hyperplasia by dried blood samples.

    PubMed

    Wieacker, Isabelle; Peter, Michael; Borucki, Katrin; Empting, Susann; Roehl, Friedrich-Wilhelm; Mohnike, Klaus

    2015-07-01

    Careful monitoring of the therapy is crucial for patients with congenital adrenal hyperplasia (CAH) in order to prevent the effects of increased androgen production as well as life-threatening salt-wasting crisis. The key metabolite, 17α-hydroxyprogesterone (17-OHP) can be detected in serum, saliva or dried blood. In clinical practice there are challenges due to discomfort of venous blood sampling and complicated retrieval of saliva during infancy. Furthermore, the immunoassay method is limited in its specificity due to cross-reactions. In this observational study we prospectively examined over a period of 5 years, 20 patients with CAH due to 21-hydroxylase deficiency using standard immunoassays for serum samples (radioimmunoassay and enzyme immunoassay) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) in dried blood spots. Bland-Altman plots show goodness of agreement between both the methods for the desirable therapeutic concentration range of 17-OHP. LC-MS/MS is characterized by a high accuracy in the therapeutic concentration range of 17-OHP <100 nmol/L (r=0.91). Dried blood samples are convenient and reliable specimen for 17-OHP measured by LC-MS/MS. This method could be used for home monitoring of hydrocortisone replacement therapy both in salt-waster and simple virilizer CAH.

  13. Malaria PCR detection in Cambodian low-transmission settings: dried blood spots versus venous blood samples.

    PubMed

    Canier, Lydie; Khim, Nimol; Kim, Saorin; Eam, Rotha; Khean, Chanra; Loch, Kaknika; Ken, Malen; Pannus, Pieter; Bosman, Philippe; Stassijns, Jorgen; Nackers, Fabienne; Alipon, SweetC; Char, Meng Chuor; Chea, Nguon; Etienne, William; De Smet, Martin; Kindermans, Jean-Marie; Ménard, Didier

    2015-03-01

    In the context of malaria elimination, novel strategies for detecting very low malaria parasite densities in asymptomatic individuals are needed. One of the major limitations of the malaria parasite detection methods is the volume of blood samples being analyzed. The objective of the study was to compare the diagnostic accuracy of a malaria polymerase chain reaction assay, from dried blood spots (DBS, 5 μL) and different volumes of venous blood (50 μL, 200 μL, and 1 mL). The limit of detection of the polymerase chain reaction assay, using calibrated Plasmodium falciparum blood dilutions, showed that venous blood samples (50 μL, 200 μL, 1 mL) combined with Qiagen extraction methods gave a similar threshold of 100 parasites/mL, ∼100-fold lower than 5 μL DBS/Instagene method. On a set of 521 field samples, collected in two different transmission areas in northern Cambodia, no significant difference in the proportion of parasite carriers, regardless of the methods used was found. The 5 μL DBS method missed 27% of the samples detected by the 1 mL venous blood method, but most of the missed parasites carriers were infected by Plasmodium vivax (84%). The remaining missed P. falciparum parasite carriers (N = 3) were only detected in high-transmission areas.

  14. Malaria PCR Detection in Cambodian Low-Transmission Settings: Dried Blood Spots versus Venous Blood Samples

    PubMed Central

    Canier, Lydie; Khim, Nimol; Kim, Saorin; Eam, Rotha; Khean, Chanra; Loch, Kaknika; Ken, Malen; Pannus, Pieter; Bosman, Philippe; Stassijns, Jorgen; Nackers, Fabienne; Alipon, SweetC; Char, Meng Chuor; Chea, Nguon; Etienne, William; De Smet, Martin; Kindermans, Jean-Marie; Ménard, Didier

    2015-01-01

    In the context of malaria elimination, novel strategies for detecting very low malaria parasite densities in asymptomatic individuals are needed. One of the major limitations of the malaria parasite detection methods is the volume of blood samples being analyzed. The objective of the study was to compare the diagnostic accuracy of a malaria polymerase chain reaction assay, from dried blood spots (DBS, 5 μL) and different volumes of venous blood (50 μL, 200 μL, and 1 mL). The limit of detection of the polymerase chain reaction assay, using calibrated Plasmodium falciparum blood dilutions, showed that venous blood samples (50 μL, 200 μL, 1 mL) combined with Qiagen extraction methods gave a similar threshold of 100 parasites/mL, ∼100-fold lower than 5 μL DBS/Instagene method. On a set of 521 field samples, collected in two different transmission areas in northern Cambodia, no significant difference in the proportion of parasite carriers, regardless of the methods used was found. The 5 μL DBS method missed 27% of the samples detected by the 1 mL venous blood method, but most of the missed parasites carriers were infected by Plasmodium vivax (84%). The remaining missed P. falciparum parasite carriers (N = 3) were only detected in high-transmission areas. PMID:25561570

  15. A study of blood alcohol stability in forensic antemortem blood samples.

    PubMed

    Shan, Xiaoqin; Tiscione, Nicholas B; Alford, Ilene; Yeatman, Dustin Tate

    2011-09-10

    The effect of long-term storage on alcohol stability in preserved forensic antemortem blood samples was investigated. Thirty-two whole blood case samples (each with two tubes of blood) were used for this study. One tube from each case was analyzed for blood alcohol concentration (BAC) for court proceedings of driving under the influence (DUI), and all blood samples were then stored under refrigeration. After the storage time (ranging from 13 to 39 months) both tubes of blood for each case were reanalyzed for BAC and the results were compared to the original analysis. Seven samples originally negative for alcohol analysis remained negative. The comparative data for 25 samples demonstrated various losses in BAC in both tubes. A significant loss with a mean of 0.015g/dL, was observed in previously opened tubes compared to a mean loss of 0.010g/dL in unopened tubes. In order to determine the effect of other storage conditions, the same blood samples were then stored at room temperature for 6 months followed by 38°C for 7 and 28 days and analyzed for BAC at the end of each storage time period. The seven alcohol negative cases remained negative when stored at room temperature or at 38°C. Six months of storage at room temperature decreased BAC further for both tubes of the alcohol positive cases with a mean loss of 0.014g/dL. Further storage at 38°C for 7 days did not cause any significant change in BAC. Storage at 38°C for 28 days caused some loss in BAC which was determined to be significant by statistical analysis.

  16. Vesicle-associated microRNAs are released from blood cells on incubation of blood samples.

    PubMed

    Köberle, Verena; Kakoschky, Bianca; Ibrahim, Ahmed Atef; Schmithals, Christian; Peveling-Oberhag, Jan; Zeuzem, Stefan; Kronenberger, Bernd; Waidmann, Oliver; Pleli, Thomas; Piiper, Albrecht

    2016-03-01

    MicroRNAs (miRNAs) circulating extracellularly in the blood are currently intensively studied as novel disease markers. However, the preanalytical factors influencing the levels of the extracellular miRNAs are still incompletely explored. In particular, it is unknown, whether the incubation of blood samples as occurring in clinical routine can lead to a release of miRNAs from blood cells and thus alter the extracellular miRNA levels before the preparation of serum or plasma from the blood cells. Using a set of marker miRNAs and quantitative RT-PCR, we found that the levels of extracellular miRNA-1, miRNA-16, and miRNA-21 were increased in EDTA and serum collection tubes incubated for 1-3 hours at room temperature and declined thereafter; the levels of the liver-specific miRNA-122 declined monophasically. These events occurred in the absence of significant hemolysis. When the blood was supplemented with Ribonuclease A inhibitor, the levels of miRNA-1, miRNA-16, and miRNA-21 increased substantially during the initial 3 hours of incubation and those of miRNA-122 remained unchanged, indicating that the release of blood cell-derived miRNAs occurred during the initial 3 hours of incubation of the blood tubes, but not at later time points. Separation of 5-hour preincubated blood into vesicle and nonvesicle fractions revealed a selective increase in the portion of vesicle-associated miRNAs. Together, these data indicate that the release of vesicle-associated miRNAs from blood cells can occur in blood samples within the time elapsing in normal clinical practice until their processing without significant hemolysis. This becomes particularly visible on the inhibition of miRNA degradation by Ribonuclease A inhibitor.

  17. Foetal blood sampling. Practical approach to management of foetal distress.

    PubMed

    Coltart, T M; Trickey, N R; Beard, R W

    1969-02-08

    The practical application of foetal blood sampling in the routine management of patients in labour has been reviewed in a six-month survey, during which time 1,668 patients were delivered at Queen Charlotte's Hospital.Foetal acidaemia (pH 7.25 or less) occurred in 45 of the 295 patients who showed clinical signs of foetal distress. Foetal tachycardia was the presenting sign in 33 of these 45 patients, underlining the importance of this physical sign. Foetal acidaemia in association with clinical foetal distress occurred twice as often in patients who had complications of pregnancy and who were therefore regarded as obstetrically "at risk" as it did in patients who were obstetrically "normal" No cases of acidaemia were detected in any of the foetal blood samples performed routinely on "at-risk" patients in the absence of clinical foetal distress.

  18. Comparison of Proteins in Whole Blood and Dried Blood Spot Samples by LC/MS/MS

    NASA Astrophysics Data System (ADS)

    Chambers, Andrew G.; Percy, Andrew J.; Hardie, Darryl B.; Borchers, Christoph H.

    2013-09-01

    Dried blood spot (DBS) sampling methods are desirable for population-wide biomarker screening programs because of their ease of collection, transportation, and storage. Immunoassays are traditionally used to quantify endogenous proteins in these samples but require a separate assay for each protein. Recently, targeted mass spectrometry (MS) has been proposed for generating highly-multiplexed assays for biomarker proteins in DBS samples. In this work, we report the first comparison of proteins in whole blood and DBS samples using an untargeted MS approach. The average number of proteins identified in undepleted whole blood and DBS samples by liquid chromatography (LC)/MS/MS was 223 and 253, respectively. Protein identification repeatability was between 77 %-92 % within replicates and the majority of these repeated proteins (70 %) were observed in both sample formats. Proteins exclusively identified in the liquid or dried fluid spot format were unbiased based on their molecular weight, isoelectric point, aliphatic index, and grand average hydrophobicity. In addition, we extended this comparison to include proteins in matching plasma and serum samples with their dried fluid spot equivalents, dried plasma spot (DPS), and dried serum spot (DSS). This work begins to define the accessibility of endogenous proteins in dried fluid spot samples for analysis by MS and is useful in evaluating the scope of this new approach.

  19. Molecular recognition of HER-1 in whole-blood samples.

    PubMed

    Moldoveanu, Iuliana; Stanciu Gavan, Camelia; Stefan-van Staden, Raluca-Ioana

    2014-11-01

    Multimode sensing was proposed for molecular screening and recognition of HER-1 in whole blood. The tools used for molecular recognition were platforms based on nanostructured materials such as the complex of Mn(III) with meso-tetra (4-carboxyphenyl) porphyrin, and maltodextrin (dextrose equivalence between 4 and 7), immobilized in diamond paste, graphite paste or C60 fullerene paste. The identification of HER-1 in whole-blood samples, at molecular level, is performed using stochastic mode and is followed by the quantification of it using stochastic and differential pulse voltammetry modes. HER-1 can be identified in the concentration range between 280 fg/ml and 4.86 ng/ml using stochastic mode, this making possible the early detection of cancers such as gastrointestinal, pancreatic and lung cancers. The recovery tests performed using whole-blood samples proved that the platforms can be used for identification and quantification of HER-1 with high sensitivity and reliability in such samples, these making them good molecular screening tools. Copyright © 2014 John Wiley & Sons, Ltd.

  20. Characterization of differences between blood sample matrices in untargeted metabolomics.

    PubMed

    Denery, Judith R; Nunes, Ashlee A K; Dickerson, Tobin J

    2011-02-01

    Large-scale proteomic and metabolomic technologies are increasingly gaining attention for their use in the diagnosis of human disease. In order to ensure the statistical power of relevant markers, such analyses must incorporate a large number of representative samples. While in a best-case scenario these samples are collected through a study design that is specifically tailored for the desired analysis, often studies must rely upon the analysis of large numbers of previously banked samples that may or may not have complete and accurate documentation of their associated collection and storage methods. In this study, several human blood matrices were analyzed and compared for the quality of metabolomic output. The sample types that were tested include plasma prepared with a variety of anticoagulants and serum collected by venipuncture and capillary blood collection protocols. Analysis with liquid chromatography-mass spectrometry (LC-MS) revealed only subtle differences between the various plasma preparation methods. Differences between the serum and plasma samples appear to be largely peptide/protein-based and are consistent with the biological distinction of the two matrices. Interestingly, the small molecule lysophosphatidylinositol was found to be in higher abundance in plasma, as a possible consequence of the effect of the intrinsic clotting cascade on adjacent metabolic pathways. Comparison of the small-molecule profiles of the capillary- and venipuncture-collected samples revealed 23 statistically significant compound differences between these sample types. Most of these features can be attributed to surfactants and detergents used to pretreat the skin in order to maintain the sterility of sample collection. However, several have identical mass and molecular formulas as endogenous human metabolites and could be erroneously attributed to actual metabolic perturbations. Understanding the extent of these matrix effects is important for control of systematic bias

  1. Appropriate timing of blood sampling for blood gas analysis in the ventilated rabbit.

    PubMed

    Sei, Kiguna; Fujita, Masanori; Okawa, Shinpei; Hirasawa, Takeshi; Kushibiki, Toshihiro; Sasa, Hidenori; Furuya, Kenichi; Ishihara, Miya

    2016-12-01

    Arterial and venous blood gas analyses (BGAs) are essential to evaluate devices that measure biological oxygenation. The appropriate timing of blood sampling for BGA after respiratory rate (RR) change in animal experiments has not been reported. This study investigated the appropriate timing of blood sampling for BGA in ventilated rabbits and whether venous samples are an alternative to arterial samples. Under general anesthesia, 14 rabbits (body weight, 3.02 ± 0.09 kg) were ventilated and their RR was changed (40/min, 30/min, and 20/min). Blood was sampled through cervical arterial and venous catheters. Experiment 1: in seven rabbits, arterial BGA was measured at 0, 0.5, 1, 2, 3, 5, 10, 15, and 20 min after the RR change. Experiment 2: in seven different rabbits, simultaneous arterial and venous BGA were measured at 0, 2, 5, 10, 15, and 20 min after the RR change. Oxygen partial pressure (PO2) and saturation (SO2) of the arterial blood stabilized 0.5 min after the RR changed. In venous BGA, no index stabilized during observation. The arterial and venous values of the carbon dioxide partial pressure (PCO2) and pH had significant correlations (arterial PCO2 = 0.9316 × venous PCO2-4.4425 [r = 0.9178]; arterial pH = 1.0835 × venous pH-0.5795 [r = 0.9453]). In ventilated rabbits, arterial PO2 and SO2 stabilized in 0.5 min. No venous value stabilized after the RR change. Only the PCO2 and pH of venous samples may be an alternative to arterial samples under the defined formula. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

  2. On-chip sample preparation for complete blood count from raw blood.

    PubMed

    Nguyen, John; Wei, Yuan; Zheng, Yi; Wang, Chen; Sun, Yu

    2015-03-21

    This paper describes a monolithic microfluidic device capable of on-chip sample preparation for both RBC and WBC measurements from whole blood. For the first time, on-chip sample processing (e.g. dilution, lysis, and filtration) and downstream single cell measurement were fully integrated to enable sample preparation and single cell analysis from whole blood on a single device. The device consists of two parallel sub-systems that perform sample processing and electrical measurements for measuring RBC and WBC parameters. The system provides a modular environment capable of handling solutions of various viscosities by adjusting the length of channels and precisely controlling mixing ratios, and features a new 'offset' filter configuration for increased duration of device operation. RBC concentration, mean corpuscular volume (MCV), cell distribution width, WBC concentration and differential are determined by electrical impedance measurement. Experimental characterization of over 100,000 cells from 10 patient blood samples validated the system's capability for performing on-chip raw blood processing and measurement.

  3. Long-Term Blood Alcohol Stability in Forensic Antemortem Whole Blood Samples.

    PubMed

    Tiscione, Nicholas B; Vacha, Ruth E; Alford, Ilene; Yeatman, Dustin Tate; Shan, Xiaoqin

    2015-01-01

    The effect of long-term room temperature storage on the stability of ethanol in whole blood specimens was investigated. One hundred and seventeen preserved whole blood case samples (110 of 117 with two tubes of blood in each case) were used for this study. One tube from each case was initially tested for blood alcohol concentration (BAC) for criminal driving under the influence proceedings. Cases positive for ethanol ranged in BAC from 0.023 to 0.281 g/dL. The second tube, if present, remained sealed. All blood samples were then stored at room temperature. After 5.4-10.3 years, the opened tubes were reanalyzed for BAC by the same laboratory that performed the initial testing using the same method and same instrumentation. After the same storage period, the unopened tubes were sent to a different laboratory, using a different method and different instrumentation, and reanalyzed for BAC after a total of 5.6-10.5 years of room temperature storage. Seven samples initially negative for alcohol remained negative. All samples initially positive for ethanol demonstrated a decrease in BAC over time with a statistically significant difference in loss observed based on blood sample volume and whether or not the tube had been previously opened. The decrease in BAC ranged from 0.005 to 0.234 g/dL. Tubes that were not previously opened and were more than half full demonstrated better BAC stability with 89% of these tubes demonstrating a loss of BAC between 0.01 and 0.05 g/dL.

  4. Guidance for storing blood samples in laboratories performing complete blood count with differential.

    PubMed

    Cornet, E; Behier, C; Troussard, X

    2012-12-01

    The complete blood count (CBC) with differential leukocyte count (DIFF) is an important part of clinical laboratory analyses and provides crucial data for clinicians. Delivery time after blood collection and conditions of storage is known to affect the reliability of results of some hematologic parameters. The aim of this study was to assess the variations of hematologic parameters over time and the influence of storage temperature. Blood samples were randomly selected from hospitalized patients and stored at room temperature and at 4 °C. CBC and DIFF were performed on an automated hematology analyzer and the results between the two groups were compared. Samples stored at room temperature showed an important increase in mean corpuscular volume and hematocrit and a decrease in mean corpuscular hemoglobin concentration. Neutrophil counts tended to increase, whereas monocyte counts tended to decrease. Storing samples at 4 °C improved reproducibility over time of all quantitative and qualitative parameters. We also observed that NEUT-X, a routine parameter useful in detecting myelodysplastic syndrome, became unreliable when analyzed 24 h after sample collection. Our results led us to recommend that samples should be analyzed within 6 h, particularly if samples are transported at room temperature. We also recommend storing samples at 4 °C in case of remote CBC analysis, especially in the context of clinical trials. © 2012 Blackwell Publishing Ltd.

  5. Development of blood extraction system designed by female mosquito's blood sampling mechanism for bio-MEMS

    NASA Astrophysics Data System (ADS)

    Tsuchiya, Kazuyoshi; Nakanishi, Naoyuki; Nakamachi, Eiji

    2005-02-01

    A compact and wearable wristwatch type Bio-MEMS such as a health monitoring system (HMS) to detect blood sugar level for diabetic patient, was newly developed. The HMS consists of (1) a indentation unit with a microneedle to generate the skin penetration force using a shape memory alloy(SMA) actuator, (2) a pumping unit using a bimorph PZT piezoelectric actuator to extract the blood and (3) a gold (Au) electrode as a biosensor immobilized GOx and attached to the gate electrode of MOSFET to detect the amount of Glucose in extracted blood. GOx was immobilized on a self assembled spacer combined with an Au electrode by the cross-link method using BSA as an additional bonding material. The device can extract blood in a few microliter through a painless microneedle with the negative pressure by deflection of the bimorph PZT piezoelectric actuator produced in the blood chamber, by the similar way the female mosquito extracts human blood with muscle motion to flex or relax. The performances of the liquid sampling ability of the pumping unit through a microneedle (3.8mm length, 100μm internal diameter) using the bimorph PZT piezoelectric microactuator were measured. The blood extraction micro device could extract human blood at the speed of 2μl/min, and it is enough volume to measure a glucose level, compared to the amount of commercial based glucose level monitor. The electrode embedded in the blood extraction device chamber could detect electrons generated by the hydrolysis of hydrogen peroxide produced by the reaction between GOx and glucose in a few microliter extracted blood, using the constant electric current measurement system of the MOSFET type hybrid biosensor. The output voltage for the glucose diluted in the chamber was increased lineally with increase of the glucose concentration.

  6. Bacterial and fungal DNA extraction from blood samples: manual protocols.

    PubMed

    Lorenz, Michael G; Mühl, Helge; Disqué, Claudia

    2015-01-01

    A critical point of molecular diagnosis of systemic infections is the method employed for the extraction of microbial DNA from blood. A DNA isolation method has to be able to fulfill several fundamental requirements for optimal performance of diagnostic assays. First of all, low- and high-molecular-weight substances of the blood inhibitory to downstream analytical reactions like PCR amplification have to be removed. This includes human DNA which is a known source of false-positive results and factor decreasing the analytical sensitivity of PCR assays by unspecific primer binding. At the same time, even extremely low amounts of microbial DNA need to be supplied to molecular diagnostic assays in order to detect low pathogen loads in the blood. Further, considering the variety of microbial etiologies of sepsis, a method should be capable of lysing Gram-positive, Gram-negative, and fungal organisms. Last, extraction buffers, reagents, and consumables have to be free of microbial DNA which leads to false-positive results. Here, we describe manual methods which allow the extraction of microbial DNA from small- and large-volume blood samples for the direct molecular analysis of pathogen.

  7. The safety of brachial artery puncture for arterial blood sampling.

    PubMed

    Okeson, G C; Wulbrecht, P H

    1998-09-01

    This study was designed to determine the incidence of complications in a sample of 6,185 brachial artery punctures for arterial blood gas analysis. The study sample was comprised of adult patients who had arterial blood gas analysis ordered in the course of their clinical evaluations in a multispecialty clinic and hospital affiliated with a university school of medicine. Subjects were entered prospectively at the time the procedure was done. The overall incidence of all complications was 2.0%. Immediate limb pain or parenthesias occurred in 1.1%, while the onset of symptoms was delayed up to 24 h in 0.9%. Hematoma formation occurred in only 0.06%. None of the complications was considered to be of major impact, in that none was associated with limb ischemia or other objective abnormalities. Only one subject required analgesic medication to control pain that ultimately subsided spontaneously without deficit. We believe that brachial artery puncture, when properly performed, is a safe and reliable alternative route for obtaining arterial blood for gas analysis.

  8. Nanoliter viscometer for analyzing blood plasma and other liquid samples.

    PubMed

    Srivastava, Nimisha; Davenport, Robertson D; Burns, Mark A

    2005-01-15

    We have developed a microfabricated nanoliter capillary viscometer that quickly, easily, and inexpensively measures the viscosity of liquids. The measurement of viscosity is based on capillary pressure-driven flow inside microfluidic channels (depth approximately 30 microm and width approximately 300 microm). Accurate and precise viscosity measurements can be made in less than 100 s while using only 600 nL of liquid sample. The silicon-glass hybrid device (18 mm by 15 mm) contains on-chip components that measure the driving capillary pressure difference and the relevant geometrical parameters; these components make the nanoliter viscometer completely self-calibrating, robust, and easy to use. Several different microfabricated viscometers were tested using solutions with viscosities ranging from 1 to 5 cP, a range relevant to biological fluids (urine, blood, blood plasma, etc.). Blood plasma samples collected from patients with the symptoms of hyperviscosity syndrome were tested on the nanoliter capillary viscometer to an accuracy of 3%. Such self-calibrating nanoliter viscometers may have widespread applications in chemical, biological, and medical laboratories as well as in personal health care.

  9. Haemolysis in neonatal blood samples: a survey of practice.

    PubMed

    Joshi, S; Vaitkute, R; Jeffery, J; Ayling, R M

    2007-03-01

    To survey how laboratories report, and neonatologists perceive them to report, the presence of haemolysis in neonatal blood samples. A questionnaire was sent to clinical biochemists and neonatologists in 88 hospitals believed to be using Roche methodology. A 49% response rate was obtained. Seventeen hospitals were excluded from analysis owing to incomplete questionnaires or altered methodology. Of the remaining 71 hospitals, 30 (42%) laboratories and 18 (25%) neonatologists responded - where both responded, none gave identical replies. Eighteen laboratories admitted the use of serum indices to make decisions on haemolysed samples. Although recommended interference limits are available from Roche, there was little consensus between laboratories. Laboratories should have clear guidelines on the processing of haemolysed samples from neonates and these should be adequately communicated to those responsible for their direct care.

  10. Experience of fetal scalp blood sampling during labor.

    PubMed

    Liljeström, Lena; Wikström, Anna-Karin; Skalkidou, Alkistis; Akerud, Helena; Jonsson, Maria

    2014-01-01

    Fetal scalp blood sampling (FBS) is often claimed to be painful for women in labor and difficult for obstetricians to perform. Our aim was to assess women's experience of pain during FBS and obstetricians' experience of difficulty in performing the test. At a tertiary center in Sweden, a questionnaire with answers on a 10-point scale was completed by 51 women and the obstetricians performing the test. Women's experience of pain had a median of 3.5. FBS was well tolerated in women who had epidural analgesia but might be associated with pain in women without. Higher maternal body mass index and less cervical dilation were associated with higher pain ratings. Obstetricians did not generally experience scalp sampling as difficult to perform (median score 3.0). However, the sampling procedure can be more complicated in situations with higher maternal body mass index, less cervical dilation, and a higher station of the fetal head.

  11. Kinetics of ethanol degradation in forensic blood samples.

    PubMed

    Ferrari, L A; Triszcz, J M; Giannuzzi, L

    2006-09-12

    To determine ethanol in human post-mortem blood samples is problematic, largely due to the inappropriate and variable methods of preserving and storing, which can cause decomposition and loss of alcohol concentration. In this study, four crucial parameters of sample conservation were studied: temperature (T), percentage of air chamber in container (%CA), ethanol concentration in blood and post-mortem time. Blood samples from post-mortem cases were stored under different conditions (ethanol levels were known in all cases); factorial design variables: (%CA) 0, 5, 20, 35, 65%; storage temperature: 25, 4 and -10 degrees C; in a total of 15 experiments. No preserving agent was used in samples. Quantification of ethanol in blood was carried out by gas chromatography with head-space FID detector. Initial ethanol concentration ranged from 0.50 to 4.30 g/L. The kinetics of degradation observed was pseudo-first-order. The parameter that characterised the kinetics of ethanol degradation (k(0)) ranged from (4 x 10(-4) and 5.0 x 10(-1) day(-1)), depending on storage conditions. A strong dependence between ethanol degradation and the content of the air chamber was observed and this dependence was found to be stronger than that between degradation and temperature; there was an experimental relation between (k(0)) and (%CA). Activation energy for different conditions, i.e. 0, 5, 20, 35 and 65 (%CA), were calculated and contour plots were made. A mathematical equation relating air chamber, temperature and ethanol concentration at a certain time was determined. This equation allowed estimation of initial concentrations of ethanol with minimal error. A good correlation between experimental data and data calculated with the equation was obtained (r(2) = 0.9998). The best storage conditions were: 0% CA and storage at -10 degrees C, obtaining an ethanol degradation of 0.01% after 15 days. However, 33% of ethanol degradation was obtained with 35% CA at 25 degrees C after 15 days. This

  12. Sex identification of polar bears from blood and tissue samples

    USGS Publications Warehouse

    Amstrup, Steven C.; Garner, G.W.; Cronin, M.A.; Patton, J.C.

    1993-01-01

    Polar bears (Ursus maritimus) can be adversely affected by hunting and other human perturbations because of low population densities and low reproduction rates. The sustainable take of adult females may be as low as 1.5% of the population. Females and accompanying young are most vulnerable to hunting, and hunters have not consistently reported the sex composition of the harvest, therefore a method to confirm the sexes of polar bears harvested in Alaska is needed. Evidence of the sex of harvested animals is often not available, but blood or other tissue samples often are. We extracted DNA from tissue and blood samples, and amplified segments of zinc finger (ZFX and ZFY) genes from both X and Y chromosomes with the polymerase chain reaction. Digestion of amplified portions of the X chromosome with the restriction enzyme HaeIII resulted in subdivision of the original amplified segment into four smaller fragments. Digestion with HaeIII did not subdivide the original segment amplified from the Y chromosome. The differing fragment sizes produced patterns in gel electrophoresis that distinguished samples from male and female bears 100% of the time. This technique is applicable to the investigation of many wildlife management and research questions.

  13. Blood transfusion in a random sample of hospitals in France.

    PubMed

    Mathoulin-Pélissier, S; Salmi, L R; Verret, C; Demoures, B

    2000-09-01

    Representative information on blood use is scarce. A large-scale study of blood recipients and blood use in France was conducted. Based on a random sampling, this study was carried out in teaching and other hospitals between March and December 1997. In each hospital, a patient was included if he or she received an allogeneic or an autologous transfusion during the observation period for that hospital. For each recipient, product and patient characteristics for 24 hours after inclusion were collected. From the 175 hospitals that had given a transfusion to at least one patient during the observation period, 3206 patients were included. Most transfusion recipients (57%) were over 65 years old; 42 percent were in teaching hospitals and 53 percent in medical wards. Among the 3044 adults, 91 percent received an allogeneic transfusion. Fifty-three percent of allogeneic units were WBC reduced. The indications most frequently reported for allogeneic transfusion were neoplasms (48%) and those for autologous transfusion were disorders of musculoskeletal (63%) or circulatory (15%) systems. The patients in nonteaching hospitals were more often transfused during surgery and were more likely to be aged and to have a musculoskeletal disorder than were patients in teaching hospitals. General collection of such data, within a system of traceability, could provide relevant denominators from which to interpret adverse-reaction data.

  14. Toxoplasma polymerase chain reaction on experimental blood samples.

    PubMed

    Joss, A W; Chatterton, J M; Evans, R; Ho-Yen, D O

    1993-01-01

    A two-stage polymerase chain reaction (PCR) assay employing oligonucleotide primers from the B1 gene of Toxoplasma gondii was developed and assessed for sensitivity and specificity. It was able to detect T. gondii DNA from as little as one parasite/sample in mock-infected rat or mouse leucocyte preparations. Parasitaemia was also identified in animals at five stages between 16 and 66 h after infection with the virulent RH strain, and at 12 stages between 2 and 38 days after infection with the cyst-forming Beverley strain. In the latter case, PCR was more sensitive than animal culture. No cross-reactions were observed in samples containing various opportunist pathogens which may also be found in the blood of immunocompromised patients.

  15. On the improvement of blood sample collection at clinical laboratories.

    PubMed

    Grasas, Alex; Ramalhinho, Helena; Pessoa, Luciana S; Resende, Mauricio G C; Caballé, Imma; Barba, Nuria

    2014-01-09

    Blood samples are usually collected daily from different collection points, such hospitals and health centers, and transported to a core laboratory for testing. This paper presents a project to improve the collection routes of two of the largest clinical laboratories in Spain. These routes must be designed in a cost-efficient manner while satisfying two important constraints: (i) two-hour time windows between collection and delivery, and (ii) vehicle capacity. A heuristic method based on a genetic algorithm has been designed to solve the problem of blood sample collection. The user enters the following information for each collection point: postal address, average collecting time, and average demand (in thermal containers). After implementing the algorithm using C programming, this is run and, in few seconds, it obtains optimal (or near-optimal) collection routes that specify the collection sequence for each vehicle. Different scenarios using various types of vehicles have been considered. Unless new collection points are added or problem parameters are changed substantially, routes need to be designed only once. The two laboratories in this study previously planned routes manually for 43 and 74 collection points, respectively. These routes were covered by an external carrier company. With the implementation of this algorithm, the number of routes could be reduced from ten to seven in one laboratory and from twelve to nine in the other, which represents significant annual savings in transportation costs. The algorithm presented can be easily implemented in other laboratories that face this type of problem, and it is particularly interesting and useful as the number of collection points increases. The method designs blood collection routes with reduced costs that meet the time and capacity constraints of the problem.

  16. On the improvement of blood sample collection at clinical laboratories

    PubMed Central

    2014-01-01

    Background Blood samples are usually collected daily from different collection points, such hospitals and health centers, and transported to a core laboratory for testing. This paper presents a project to improve the collection routes of two of the largest clinical laboratories in Spain. These routes must be designed in a cost-efficient manner while satisfying two important constraints: (i) two-hour time windows between collection and delivery, and (ii) vehicle capacity. Methods A heuristic method based on a genetic algorithm has been designed to solve the problem of blood sample collection. The user enters the following information for each collection point: postal address, average collecting time, and average demand (in thermal containers). After implementing the algorithm using C programming, this is run and, in few seconds, it obtains optimal (or near-optimal) collection routes that specify the collection sequence for each vehicle. Different scenarios using various types of vehicles have been considered. Unless new collection points are added or problem parameters are changed substantially, routes need to be designed only once. Results The two laboratories in this study previously planned routes manually for 43 and 74 collection points, respectively. These routes were covered by an external carrier company. With the implementation of this algorithm, the number of routes could be reduced from ten to seven in one laboratory and from twelve to nine in the other, which represents significant annual savings in transportation costs. Conclusions The algorithm presented can be easily implemented in other laboratories that face this type of problem, and it is particularly interesting and useful as the number of collection points increases. The method designs blood collection routes with reduced costs that meet the time and capacity constraints of the problem. PMID:24406140

  17. Stunting is associated with blood lead concentration among Bangladeshi children aged 2-3 years.

    PubMed

    Gleason, Kelsey M; Valeri, Linda; Shankar, A H; Hasan, Md Omar Sharif Ibne; Quamruzzaman, Quazi; Rodrigues, Ema G; Christiani, David C; Wright, Robert O; Bellinger, David C; Mazumdar, Maitreyi

    2016-11-04

    Lead toxicity is of particular public health concern given its near ubiquitous distribution in nature and established neurotoxicant properties. Similar in its ubiquity and ability to inhibit neurodevelopment, early childhood stunting affects an estimated 34 % of children under 5 in low- and middle-income countries. Both lead and stunting have been shown to be associated with decreased neurodevelopment, although the relationship between these childhood burdens is underexplored. The association between lead exposure and stunting has been previously established, yet limited data are available on susceptibility windows. Whole blood lead samples were collected from rural Bangladeshi children at delivery (umbilical cord blood) and at age 20-40 months (fingerstick blood). Stunting was determined using the Child Growth Standards developed from the World Health Organization Multicentre Growth Reference Study. Children with height for age < -2 z-scores below the median of the WHO Child Growth Standards were classified as stunted in all analyses. Median (IQR) umbilical cord and fingerstick blood lead levels were 3.1 (1.6-6.3) μg/dl and 4.2 (1.7-7.6) μg/dl, respectively. In adjusted multivariable regression models, the odds of stunting at 20-40 months increased by 1.12 per μg/dl increase in blood lead level (OR = 1.12, 95 % CI: 1.02-1.22). No association was found between cord blood lead level and risk of stunting (OR = 0.97, 95 % CI: 0.94-1.00). There is a significant association between stunting and concurrent lead exposure at age 20-40 months. This association is slightly attenuated after controlling for study clinic site. Additional research including more precise timing of lead exposure during these critical 20-40 months is needed.

  18. Prevention of hemolysis in blood samples collected from intravenous catheters.

    PubMed

    Lippi, Giuseppe; Avanzini, Paola; Cervellin, Gianfranco

    2013-05-01

    Samples drawn through intravenous catheters are frequently hemolyzed. We planned a prospective, randomized study to establish whether hemolysis in samples drawn from intravenous catheters may be reduced using S-Monovette® tubes collected by manual aspiration as compared with standard vacuum tubes. We studied 52 consecutive patients admitted to the ED. Blood was drawn through a 20-gauge intravenous catheter. A 5.0mL, Becton Dickinson Vacutainer® SST II Plus serum tube was collected and discarded. In the odd group of patients (i.e., n. 1, 3, 5, etc.), a second SST II tube was drawn with vacuum ("BD-V"), followed by a 5.5mL S-Monovette® serum tube collected with manual aspiration ("SD-A") and an identical S-Monovette collected by vacuum ("SD-V"). In the pair group of patients (i.e., n. 2, 4, 6, etc.), the sequence was modified to "SD-A", "SD-V" and "BD-V". Serum was separated and tested for lactate dehydrogenase (LDH), potassium and cell-free hemoglobin. The mean concentration of potassium (+2.7% in BD-V and +1.7% in SD-V, respectively), LDH (+15% in BD-V and +7% in SD-V, respectively) and cell-free hemoglobin was significantly increased when samples were collected with vacuum tubes as compared with manual aspiration. No significant differences were observed between SD-V and BD-V. The frequency of hemolyzed samples was higher when blood was collected with the vacuum as compared with SD-A (i.e., 2%), but did not differ between BD-V and SD-V (i.e., 29 versus 31%; p=0.70). S-Monovette can be used with vacuum or aspiration collection. This latter approach allows blood drawing with limited shear stress and less likelihood of generating spuriously hemolysis. Copyright © 2013 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  19. Perfluoroalkyl acids in blood serum samples from children in Taiwan.

    PubMed

    Bao, Jia; Lee, Yungling Leo; Chen, Pau-Chung; Jin, Yi-He; Dong, Guang-Hui

    2014-06-01

    Severe perfluoroalkyl acid (PFAA) contamination resulting from the fast-growing semiconductor, electrochemical, and optoelectronic industries has been determined in the river water in the vicinity of the Taipei area, Taiwan, during recent years. However, little is known about body burdens of the PFAA contaminations in local residents, especially children living in the Taipei area recently. In this study, ten target PFAA analytes consisted of three perfluorosulfonates (PFSAs) and seven perfluorocarboxylates (PFCAs) in the blood serum samples, collected from 225 healthy children with an average age of 13.6 years in the Taipei area from 2009 to 2010, were analyzed via high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). As the dominant PFAA contaminant in the blood serum samples from Taiwanese children, perfluorooctane sulfonate (PFOS) contributed 86% of all the target PFAA analytes, while the other nine analytes contributed less than 5% individually. PFOS showed the highest median up to 29 ng/mL, ranging from 0.03 to 148 ng/mL, which was higher than that observed in the serum samples collected from Taiwanese children between 2006 and 2008. Statistically, serum concentrations of perfluorobutane sulfonate (PFBS), perfluorohexane sulfonate (PFHxS), and perfluorooctanoic acid (PFOA) had significantly positive correlations with ages of children (p < 0.05). Furthermore, serum PFBS, PFHxS, and PFOA concentrations in the male children were considerably higher than those in the female children (p = 0.049, p = 0.000, p = 0.000).

  20. Flow cytometric determination of residual white blood cell levels in preserved samples from leukoreduced blood products.

    PubMed

    Palmer, Douglas S; Birch, Paul; O'Toole, Joan; Henderson, Deborah; Scalia, Vito

    2008-01-01

    In preparation for a proposed consolidated testing service, Canadian Blood Services undertook the evaluation of a commercial test kit for the enumeration by flow cytometry of residual white blood cells (rWBCs) present in preserved samples recovered from leukoreduced (LR) blood and platelet products. The stability of preserved WBCs, the equivalency of WBCs used for spiking, test method precision, specificity, reliability, accuracy, and sensitivity were investigated. For comparative purposes, WBC counts were also determined by Nageotte as well as by flow cytometry. WBCs were stable up to 4 weeks at room temperature for all components by either method. Within methods, no differences were observed due to the source of WBC used for spiking purposes. By either method, test precision was acceptable (<20% coefficient of variation) and of similar reliability at a target value of 10 +/- 5 WBCs per microL. The flow cytometric method was shown to be more specific and accurate than the Nageotte method. Sensitivity by either method was 0.1 WBCs per microL. On average, Nageotte counts were lower than those observed by flow cytometry. These results demonstrate that WBCs in WBC stabilizing solution-treated samples from LR blood components were stabilized up to 4 weeks at room temperature and that rWBC determinations made with a WBC enumeration kit by flow cytometry have the required precision, specificity, reliability, and accuracy in the relevant test range. This validated WBC stabilization and flow cytometric counting method is considered acceptable as part of a quality control program for leukoreduced blood products.

  1. Sampling and storage conditions influencing the measurement of parathyroid hormone in blood samples: a systematic review.

    PubMed

    Hanon, Elodie A; Sturgeon, Catharine M; Lamb, Edmund J

    2013-10-01

    Parathyroid hormone (PTH) is relatively unstable: optimisation of pre-analytical conditions, including specimen type, sampling time and storage conditions, is essential. We have undertaken a systematic review of these pre-analytical conditions. An electronic search of the PubMed, Embase, Cochrane, Centre for Research and Dissemination and Bandolier databases was undertaken. Of 5511 papers identified, 96 underwent full text review, of which 83 were finally included. At room temperature PTH was stable in ethylenediaminetetraacetic acid (EDTA) preserved whole blood for at least 24 h and in EDTA plasma for at least 48 h after venepuncture. Losses were observed in clotted blood samples after 3 h and in serum after 2 h. At 4°C PTH was more stable in EDTA plasma (at least 72 h) than serum (at least 24 h). Central venous PTH concentrations were higher than peripheral venous concentrations. In the northern hemisphere, PTH concentrations were higher in winter than summer. PTH has a circadian rhythm characterised by a nocturnal acrophase and mid-morning nadir. Data related to frozen storage of PTH (-20°C and -80°C) were limited and contradictory. We recommend that blood samples for PTH measurement should be taken into tubes containing EDTA, ideally between 10:00 and 16:00, and plasma separated within 24 h of venepuncture. Plasma samples should be stored at 4°C and analysed within 72 h of venepuncture. Particular regard must be paid to the venepuncture site when interpreting PTH concentration. Further research is required to clarify the suitability of freezing samples prior to PTH measurement.

  2. Natural antioxidants improve red blood cell "survival" in non-leukoreduced blood samples.

    PubMed

    Kucherenko, Yuliya V; Bernhardt, Ingolf

    2015-01-01

    Blood collected in an anticoagulant can be kept refrigerated in an unmodified state within 5 - 6 weeks. Oxidative damage is considered to be a one of the major factors contributing to the development of storage lesions. Lipid and membrane proteins oxidation results in changes in cation gradients that affect the cell survival. In the present study we used the natural antioxidants and ion channels blockers (L-carnosine, spermine, phloretin and their mixtures) to prolong "survival" of red blood cells (RBCs), measured as the lack of PS exposure and cell hemolysis, in the Alsever's preservative solution upon hypothermic storage. We show that the mixture of carnosine (20 mM), spermine (20 µM) and phloretin (100 µM) effectively blunted phosphatidylserine (PS) exposure, Ca(2+) accumulation and RBCs hemolysis in non-leukoreduced low (∼ 2%) hematocrit samples after 36 days of storage as well as after 1 day of post-storage incubation of the stored cells in physiological saline solution. In addition, a slight but significant decrease in PS exposure was observed in non-leukoreduced high (∼ 20%) hematocrit samples after 36 days of storage with the mixture of substances. We conclude that the use of the mixture of natural antioxidants (carnosine, spermine, and phloretin) as an additive to blood preservative solution provides better RBCs storage and "survival". © 2015 S. Karger AG, Basel.

  3. A modified serial blood sampling technique and utility of dried-blood spot technique in estimation of blood concentration: application in mouse pharmacokinetics.

    PubMed

    Kurawattimath, Vishwanath; Pocha, Krishna; Mariappan, T Thanga; Trivedi, Ravi Kumar; Mandlekar, Sandhya

    2012-03-01

    Pharmacokinetic (PK) studies in mice usually require discrete and parallel blood sampling owing to a restriction on the volume of blood that can be withdrawn. This results in dosing large number of animals and generating composite PK profile. To reduce the number of animals and generate individual animal PK profiles, we developed a serial sampling technique via tail vein bleeding in mice, in which only 20-30 μL blood was withdrawn per time point. Due to the small blood volume, a dried-blood spot (DBS) technique was applied for sample processing. The utility of this technique was demonstrated using three test compounds (amodiaquine, chloroquine and chlorthalidone), with varying degrees of blood-to-plasma partition ratios. The PK studies were carried out in male Balb/c mouse weighing 25-30 g. The compounds were administered intravenously via the saphenous vein. Blood was collected by composite (retro-orbital plexus) or serial (tail vein bleeding) sampling techniques at different time points. Blood samples were processed as blood lysate or DBS. Blood or plasma samples were analyzed by sensitive and rapid UPLC-MS/MS methods. The blood concentrations (both from blood lysate and DBS) obtained from serial sampling matched with those from composite sampling. The ratio of blood AUC to plasma AUC correlated well with the in vitro blood-to-plasma partition ratio of the compounds. The systemic clearance and volume of distribution at steady state calculated from blood or plasma AUCs were in proportion to the respective AUCs. Our results indicated that the serial sampling technique would reduce the number of animals and also compound usage, as well as improve the quality of pharmacokinetic data. Also, the serial sampling technique does not require the use of anesthesia and allows estimation of inter-animal variability in PK. A small volume serial sampling is possible due to the availability of the DBS technique.

  4. Interaction of two optically coupled whole blood samples during respiratory burst

    NASA Astrophysics Data System (ADS)

    Voeikov, Vladimir L.; Novikov, Cyril N.

    1997-05-01

    To study a possibility of interaction of two optically, but not chemically coupled samples of whole human blood the following experimental setup was used. A quartz cuvette with either nondiluted blood or saline was placed inside a glass vial. Saline diluted whole blood was poured into the vial and respiratory burst (RB) was initiated in it with phorbol ether or zymosan. Luminol-dependent chemiluminescence (LCL) was registered using liquid scintillation counter (coincident circuit off). Effect of blood placed in the cuvette upon photon emission from blood placed in the vial was evaluated. It was shown that blood of some donors consistently attenuated photon emission from the sample in which RB was induced. Blood of another group of donors enhanced photon emission from the `partner' sample. Some properties of blood taken from the cuvette after being in the contact with the sample in which RB was induced changed in comparison with the same blood that was contacting with the non-stimulated sample. Exposed blood has lost the ability to attenuate light emission from the fresh portion of blood in which RB was induced. Its own LCL in response to addition of zymoscan was different from that of the parallel sample of same blood not exposed to sample undergoing RB. These results suggest that two chemically separated but optically coupled samples of blood can interact.

  5. Air bubbles and hemolysis of blood samples during transport by pneumatic tube systems.

    PubMed

    Mullins, Garrett R; Bruns, David E

    2017-08-10

    Transport of blood samples through pneumatic tube systems (PTSs) generates air bubbles in transported blood samples and, with increasing duration of transport, the appearance of hemolysis. We investigated the role of air-bubble formation in PTS-induced hemolysis. Air was introduced into blood samples for 0, 1, 3 or 5min to form air bubbles. Hemolysis in the blood was assessed by (H)-index, lactate dehydrogenase (LD) and potassium in plasma. In an effort to prevent PTS-induced hemolysis, blood sample tubes were completely filled, to prevent air bubble formation, and compared with partially filled samples after PTS transport. We also compared hemolysis in anticoagulated vs clotted blood subjected to PTS transport. As with transport through PTSs, the duration of air bubble formation in blood by a gentle stream of air predicted the extent of hemolysis as measured by H-index (p<0.01), LD (p<0.01), and potassium (p<0.02) in plasma. Removing air space in a blood sample prevented bubble formation and fully protected the blood from PTS-induced hemolysis (p<0.02 vs conventionally filled collection tube). Clotted blood developed less foaming during PTS transport and was partially protected from hemolysis vs anticoagulated blood as indicated by lower LD (p<0.03) in serum than in plasma after PTS sample transport. Prevention of air bubble formation in blood samples during PTS transport protects samples from hemolysis. Copyright © 2017. Published by Elsevier B.V.

  6. Diagnosis of Carrion’s Disease by Direct Blood PCR in Thin Blood Smear Negative Samples

    PubMed Central

    Tinco Valdez, Carmen; Pons, Maria J.; del Valle, Luis J.; Oré, Verónica Casabona; Michelena, Denisse Champin; Mayra, Jorge Bazán; Gavidea, Víctor Zavaleta; Vargas, Martha; Ruiz, Joaquim

    2014-01-01

    Bartonella bacilliformis is the etiologic agent of Carrion's disease. This disease has two well established phases, the most relevant being the so called Oroya Fever, in which B. bacilliformis infect the erythrocytes resulting in severe anemia and transient immunosuppression, with a high lethality in the absence of adequate antibiotic treatment. The presence of B. bacilliformis was studied in 113 blood samples suspected of Carrion’s disease based on clinical criteria, despite the absence of a positive thin blood smear, by two different PCR techniques (using Bartonella-specific and universal 16S rRNA gene primers), and by bacterial culture. The specific 16S rRNA gene primers revealed the presence of 21 B. bacilliformis and 1 Bartonella elizabethae, while universal primers showed both the presence of 3 coinfections in which a concomitant pathogen was detected plus Bartonella, in addition to the presence of infections by other microorganisms such as Agrobacterium or Bacillus firmus. These data support the need to implement molecular tools to diagnose Carrion’s disease. PMID:24651298

  7. Diagnosis of Carrion's disease by direct blood PCR in thin blood smear negative samples.

    PubMed

    del Valle Mendoza, Juana; Silva Caso, Wilmer; Tinco Valdez, Carmen; Pons, Maria J; del Valle, Luis J; Oré, Verónica Casabona; Michelena, Denisse Champin; Mayra, Jorge Bazán; Gavidea, Víctor Zavaleta; Vargas, Martha; Ruiz, Joaquim

    2014-01-01

    Bartonella bacilliformis is the etiologic agent of Carrion's disease. This disease has two well established phases, the most relevant being the so called Oroya Fever, in which B. bacilliformis infect the erythrocytes resulting in severe anemia and transient immunosuppression, with a high lethality in the absence of adequate antibiotic treatment. The presence of B. bacilliformis was studied in 113 blood samples suspected of Carrion's disease based on clinical criteria, despite the absence of a positive thin blood smear, by two different PCR techniques (using Bartonella-specific and universal 16S rRNA gene primers), and by bacterial culture. The specific 16S rRNA gene primers revealed the presence of 21 B. bacilliformis and 1 Bartonella elizabethae, while universal primers showed both the presence of 3 coinfections in which a concomitant pathogen was detected plus Bartonella, in addition to the presence of infections by other microorganisms such as Agrobacterium or Bacillus firmus. These data support the need to implement molecular tools to diagnose Carrion's disease.

  8. Effect of Sample Storage Temperature and Time Delay on Blood Gases, Bicarbonate and pH in Human Arterial Blood Samples

    PubMed Central

    Mohammadhoseini, Elham; Safavi, Enayat; Seifi, Sepideh; Seifirad, Soroush; Firoozbakhsh, Shahram; Peiman, Soheil

    2015-01-01

    Background: Results of arterial blood gas analysis can be biased by pre-analytical factors, such as time interval before analysis, temperature during storage and syringe type. Objectives: To investigate the effects of samples storage temperature and time delay on blood gases, bicarbonate and PH results in human arterial blood samples. Patients and Methods: 2.5 mL arterial blood samples were drawn from 45 patients via an indwelling Intraarterial catheter. Each sample was divided into five equal samples and stored in multipurpose tuberculin plastic syringes. Blood gas analysis was performed on one of five samples as soon as possible. Four other samples were divided into two groups stored at 22°C and 0°C. Blood gas analyses were repeated at 30 and 60 minutes after sampling. Results: PaO2 of the samples stored at 0°C was increased significantly after 60 minutes (P = 0.007). The PaCO2 of the samples kept for 30 and 60 minutes at 22°C was significantly higher than primary result (P = 0.04, P < 0.001). In samples stored at 22°C, pH decreased significantly after 30 and 60 minutes (P = 0.017, P = 0.001). There were no significant differences in other results of samples stored at 0°C or 22°C after 30 or 60 minutes. Conclusions: In samples stored in plastic syringes, overestimation of PaO2 levels should be noted if samples cooled before analysis. In samples stored in plastic syringes, it is not necessary to store samples in iced water when analysis delayed up to one hour. PMID:26019892

  9. Comparisons of polybrominated diphenyl ethers levels in paired South Korean cord blood, maternal blood, and breast milk samples.

    PubMed

    Kim, Tae Hyung; Bang, Du Yeon; Lim, Hyun Jung; Won, A Jin; Ahn, Mee Young; Patra, Nabanita; Chung, Ki Kyung; Kwack, Seung Jun; Park, Kui Lea; Han, Soon Young; Choi, Wahn Soo; Han, Jung Yeol; Lee, Byung Mu; Oh, Jeong-Eun; Yoon, Jeong-Hyun; Lee, Jaewon; Kim, Hyung Sik

    2012-03-01

    Polybrominated diphenyl ethers (PBDEs), commonly used flame retardants, have been reported as potential endocrine disruptor and neurodevelopmental toxicants, thus giving rise to the public health concern. The goal of this study was to investigate the relationship between umbilical cord blood, maternal blood, and breast milk concentrations of PBDEs in South Korean. We assessed PBDE levels in paired samples of umbilical cord blood, maternal blood, and breast milk. The levels of seven PBDE congeners were measured in 21 paired samples collected from the Cheil Woman's Hospital (Seoul, Korea) in 2008. We also measured thyroid hormones levels in maternal and cord blood to assess the association between PBDEs exposure and thyroid hormone levels. However, there was no correlation between serum thyroxin (T4) and total PBDEs concentrations. The total PBDEs concentrations in the umbilical cord blood, maternal blood, and breast milk were 10.7±5.1 ng g(-1) lipid, 7.7±4.2 ng g(-1) lipid, and 3.0±1.8 ng g(-1) lipid, respectively. The ranges of total PBDE concentrations observed were 2.28-30.94 ng g(-1) lipid in umbilical cord blood, 1.8-17.66 ng g(-1) lipid in maternal blood, and 1.08-8.66 ng g(-1) lipid in breast milk. BDE-47 (45-73% of total PBDEs) was observed to be present dominantly in all samples, followed by BDE-153. A strong correlation was found for major BDE-congeners between breast milk and cord blood or maternal blood and cord blood samples. The measurement of PBDEs concentrations in maternal blood or breast milk may help to determine the concentration of PBDEs in infant.

  10. Family History Fails to Detect the Majority of Children with High Capillary Blood Total Cholesterol.

    ERIC Educational Resources Information Center

    Davidson, Dennis M.; And Others

    1991-01-01

    To examine the predictive value of family history in detecting children with high blood cholesterol, finger-stick screening was done in children (n=1,118) ages 9-10 with parental and grandparental history of cardiovascular disease and risk factors. Findings showed that screening only children with positive family histories will leave most problems…

  11. Family History Fails to Detect the Majority of Children with High Capillary Blood Total Cholesterol.

    ERIC Educational Resources Information Center

    Davidson, Dennis M.; And Others

    1991-01-01

    To examine the predictive value of family history in detecting children with high blood cholesterol, finger-stick screening was done in children (n=1,118) ages 9-10 with parental and grandparental history of cardiovascular disease and risk factors. Findings showed that screening only children with positive family histories will leave most problems…

  12. Cost Evaluation of Dried Blood Spot Home Sampling as Compared to Conventional Sampling for Therapeutic Drug Monitoring in Children.

    PubMed

    Martial, Lisa C; Aarnoutse, Rob E; Schreuder, Michiel F; Henriet, Stefanie S; Brüggemann, Roger J M; Joore, Manuela A

    2016-01-01

    Dried blood spot (DBS) sampling for the purpose of therapeutic drug monitoring can be an attractive alternative for conventional blood sampling, especially in children. This study aimed to compare all costs involved in conventional sampling versus DBS home sampling in two pediatric populations: renal transplant patients and hemato-oncology patients. Total costs were computed from a societal perspective by adding up healthcare cost, patient related costs and costs related to loss of productivity of the caregiver. Switching to DBS home sampling was associated with a cost reduction of 43% for hemato-oncology patients (€277 to €158) and 61% for nephrology patients (€259 to €102) from a societal perspective (total costs) per blood draw. From a healthcare perspective, costs reduced with 7% for hemato-oncology patients and with 21% for nephrology patients. Total savings depend on the number of hospital visits that can be avoided by using home sampling instead of conventional sampling.

  13. Cost Evaluation of Dried Blood Spot Home Sampling as Compared to Conventional Sampling for Therapeutic Drug Monitoring in Children

    PubMed Central

    Martial, Lisa C.; Aarnoutse, Rob E.; Schreuder, Michiel F.; Henriet, Stefanie S.; Brüggemann, Roger J. M.; Joore, Manuela A.

    2016-01-01

    Dried blood spot (DBS) sampling for the purpose of therapeutic drug monitoring can be an attractive alternative for conventional blood sampling, especially in children. This study aimed to compare all costs involved in conventional sampling versus DBS home sampling in two pediatric populations: renal transplant patients and hemato-oncology patients. Total costs were computed from a societal perspective by adding up healthcare cost, patient related costs and costs related to loss of productivity of the caregiver. Switching to DBS home sampling was associated with a cost reduction of 43% for hemato-oncology patients (€277 to €158) and 61% for nephrology patients (€259 to €102) from a societal perspective (total costs) per blood draw. From a healthcare perspective, costs reduced with 7% for hemato-oncology patients and with 21% for nephrology patients. Total savings depend on the number of hospital visits that can be avoided by using home sampling instead of conventional sampling. PMID:27941974

  14. Observational study to determine factors associated with blood sample haemolysis in the emergency department.

    PubMed

    Ong, Marcus E H; Chan, Yiong Huak; Lim, Chin Siah

    2008-09-01

    Haemolysis of blood samples is a common problem encountered in the Emergency department (ED). It leads to inaccurate blood results and has cost implications as blood samples very often have to be retaken. The purpose of our study was to determine which factors in blood sampling were associated with higher rates of haemolysis. An observational convenience sample of all patients presenting to the ED requiring blood urea and electrolyte (UE) analysis were eligible for our study. Questionnaires were distributed to the doctors and nurses conducting blood sampling to determine the method used and outcome data were collected after the samples were processed. Out of 227 UE samples analysed, 45 (19.8%) were haemolysed. Various factors, including method (IV cannulation or venepuncture), system (syringe or vacutainer), operator, rate of blood flow, difficulty of cannulation/venepuncture and source of blood (arterial or venous), were analysed, but their effects on haemolysis were not statistically significant (P >0.05). However, the use of the vacutainer system was associated with the highest rates of haemolysis [adjusted odds ratio (OR), 6.0; 95% confidence interval (CI), 2.3 to 15.1]. We found blood sampling with the vacutainer system to have increased rates of haemolysis. This could potentially change attitudes towards equipment used for blood sampling in the ED.

  15. Microcontroller-based system for collecting anaerobic blood samples from a running greyhound.

    PubMed

    Schmalzried, R T; Toll, P W; Devore, J J; Fedde, M R

    1992-04-01

    Many physiological variables change rapidly in the blood during sprint exercise. To characterize the dynamics and extent of these changes, blood samples must be obtained during exercise. We describe herein a portable, microcontroller-based system used to automatically obtain repeated, anaerobic, arterial blood samples from greyhounds before, during, and following a race on a track. In addition, the system also records the blood temperature in the pulmonary artery each time a blood sample is taken. The system has been tested for more than 2 years and has proven to be reliable and effective.

  16. Pediatric blood sample collection from a pre-existing peripheral intravenous (PIV) catheter.

    PubMed

    Braniff, Heather; DeCarlo, Ann; Haskamp, Amy Corey; Broome, Marion E

    2014-01-01

    Aiming to minimize pain in a hospitalized child, the purpose of this observational study was to describe characteristics of blood samples collected from pre-existing peripheral intravenous (PIV) catheters in pediatric patients. One hundred and fifty blood samples were reviewed for number of unusable samples requiring a specimen to be re-drawn. Success of the blood draw and prevalence of the loss of the PIV following blood collection was also measured. Findings included one clotted specimen, success rate of 91.3%, and 1.3% of PIVs becoming non-functional after collection. Obtaining blood specimens from a pre-existing PIV should be considered in a pediatric patient.

  17. A micro blood sampling system for catheterized neonates and pediatrics in intensive care unit.

    PubMed

    Jung, Wooseok; Ahn, Chong H

    2013-04-01

    A new micro blood sampling system has been designed, fabricated, and characterized to reduce iatrogenic blood loss from the catheterized neonates and pediatrics in intensive care unit by providing micro-volume of blood to analytical biomedical microdevices which can do point-of-care testing for their critical care. The system can not only save enormous iatrogenic blood loss through 1 to 10 μL of blood sampling and re-infusion of 1 to 5 mL of discard blood but also reduce the infection risk through the closed structure while satisfying the key criteria of the blood sampler. The sampled blood preserved its quality without rupturing of red blood cells verified by blood potassium concentrations of 3.86 ± 0.07 mM on the sampled blood which is similar to 3.81 ± 0.04 mM measured from the blood which did not go through the system. The sampling volume among the sampling channels showed consistency with the relative standard deviation of 1.41 %. In addition to the micro blood sampling capability, the sampling system showed negligible sample cross-contamination. The analyte-free samples collected after aspirating 7,500 times higher signal sample showed the same output signal as blank. The system was also demonstrated not to cause air-embolism by having no bubble generation during flushing procedure and the system was verified as leak-free since there was no fluid leakage under 30 times higher pressure than central venous pressure for 24 h.

  18. A technique for extracting blood samples from mice in fire toxicity tests

    NASA Technical Reports Server (NTRS)

    Bucci, T. J.; Hilado, C. J.; Lopez, M. T.

    1976-01-01

    The extraction of adequate blood samples from moribund and dead mice has been a problem because of the small quantity of blood in each animal and the short time available between the animals' death and coagulation of the blood. These difficulties are particularly critical in fire toxicity tests because removal of the test animals while observing proper safety precautions for personnel is time-consuming. Techniques for extracting blood samples from mice were evaluated, and a technique was developed to obtain up to 0.8 ml of blood from a single mouse after death. The technique involves rapid exposure and cutting of the posterior vena cava and accumulation of blood in the peritoneal space. Blood samples of 0.5 ml or more from individual mice have been consistently obtained as much as 16 minutes after apparent death. Results of carboxyhemoglobin analyses of blood appeared reproducible and consistent with carbon monoxide concentrations in the exposure chamber.

  19. Detection of Epstein-Barr virus and human cytomegalovirus in blood and oral samples: comparison of three sampling methods.

    PubMed

    Imbronito, Ana V; Grande, Sabrina R; Freitas, Nivea M de; Okuda, Osmar; Lotufo, Roberto F M; Nunes, Fabio D

    2008-03-01

    The purpose of this study was to identify and compare the presence of HCMV and EBV-1 in subgingival plaque, unstimulated saliva and peripheral blood of patients with chronic periodontitis. Forty patients diagnosed with chronic periodontitis (mean age, 41.7 years) were recruited. Unstimulated saliva, subgingival plaque and peripheral blood were collected from each patient and the DNA of each sample was isolated. The viruses were detected using the nested PCR technique. The detection frequency of EBV-1 in subgingival plaque, saliva and peripheral blood was 45%, 37.5% and 25%, respectively. HCMV was detected in 82.5% of subgingival plaque samples and peripheral blood and in 75% of salivary samples. The sensitivity for detecting EBV-1 in saliva and peripheral blood when EBV-1 was detected in subgingival plaque samples was low (22% and 27.7%, respectively) and the sensitivity for detecting HCMV in saliva and peripheral blood when compared to subgingival plaque was high (81.8% and 87.8%, respectively). There is a high agreement among the three sampling methods in detection of HCMV, but the detection of EBV-1 would require a combination of saliva and subgingival plaque sampling to avoid false negative results.

  20. Blood doping: potential of blood and urine sampling to detect autologous transfusion.

    PubMed

    Segura, J; Lundby, C

    2014-05-01

    The collection of blood, its storage as red blood cell (RBC) concentrates and its reinjection is prohibited; until now, the practice cannot be reliably detected. A recent innovation-the haematological module of the athlete's biological passport-can provide authorities with indications towards autologous blood transfusion. In situations where a given athlete has been exposed to altitude, heat stress, sickness, etc, additional evidence may be needed to establish beyond any reasonable doubt that a blood transfusion may actually have occurred. Additional evidence may be obtained from at least three different approaches using parameters related to blood and urine matrices.Genomics applied to mRNA or miRNA is one of the most promising analytical tools. Proteomics of changes associated with RBC membranes may reveal the presence of cells stored for some time, as can an abnormal pattern of size distribution of aged cells. In urine, high concentrations of metabolites of plasticisers originating from the blood storing bags strongly suggest a recent blood transfusion. We emphasise the usefulness of simultaneously obtaining and then analysing blood and urine for complementary evidence of autologous blood transfusion ('blood doping').

  1. Saliva samples are a viable alternative to blood samples as a source of DNA for high throughput genotyping

    PubMed Central

    2012-01-01

    Background The increasing trend for incorporation of biological sample collection within clinical trials requires sample collection procedures which are convenient and acceptable for both patients and clinicians. This study investigated the feasibility of using saliva-extracted DNA in comparison to blood-derived DNA, across two genotyping platforms: Applied Biosystems TaqmanTM and Illumina BeadchipTM genome-wide arrays. Method Patients were recruited from the Pharmacogenetics of Breast Cancer Chemotherapy (PGSNPS) study. Paired blood and saliva samples were collected from 79 study participants. The Oragene DNA Self-Collection kit (DNAgenotek®) was used to collect and extract DNA from saliva. DNA from EDTA blood samples (median volume 8 ml) was extracted by Gen-Probe, Livingstone, UK. DNA yields, standard measures of DNA quality, genotype call rates and genotype concordance between paired, duplicated samples were assessed. Results Total DNA yields were lower from saliva (mean 24 μg, range 0.2–52 μg) than from blood (mean 210 μg, range 58–577 μg) and a 2-fold difference remained after adjusting for the volume of biological material collected. Protein contamination and DNA fragmentation measures were greater in saliva DNA. 78/79 saliva samples yielded sufficient DNA for use on Illumina Beadchip arrays and using Taqman assays. Four samples were randomly selected for genotyping in duplicate on the Illumina Beadchip arrays. All samples were genotyped using Taqman assays. DNA quality, as assessed by genotype call rates and genotype concordance between matched pairs of DNA was high (>97%) for each measure in both blood and saliva-derived DNA. Conclusion We conclude that DNA from saliva and blood samples is comparable when genotyping using either Taqman assays or genome-wide chip arrays. Saliva sampling has the potential to increase participant recruitment within clinical trials, as well as reducing the resources and organisation required for multicentre sample

  2. Measurement of hematological, clinical chemistry, and infection parameters from hirudinized blood collected in universal blood sampling tubes.

    PubMed

    Menssen, H D; Brandt, N; Leben, R; Müller, F; Thiel, E; Melber, K

    2001-08-01

    Hirudin, the anticoagulatory polypeptide of the leech Hirudo medicinalis, strongly inhibits thrombus formation by specifically interacting with thrombin. For diagnostic purposes, hirudin should be superior to other anticlotting compounds because it only minimally alters the mineral, protein, and cellular blood constituents. To test this hypothesis, hirudinized and routinely processed venous blood from 80 healthy volunteers and patients was subjected to a variety of automated blood tests. A strong correlation was found between the results of automated complete blood counts obtained from K(2)-ethylenediaminetetraacetic acid (EDTA) anticoagulated and hirudinized blood (1000 antithrombin units [ATU] hirudin/ml). In addition, clinical chemistry and serological infection parameters (asparlat amintransferase [ASAT], lactate dehydrogenase [LDH], sodium, and so on, and antibodies against hepatitis B and C and human immunodeficiency virus [HIV]1/2, respectively) correlated well when measured in serum as compared with hirudinized plasma. Contrary to single clotting factors, global coagulation parameters (activated partial thromboplastin time [aPTT], prothrombin time [PT]) could not be measured in hirudinized blood. Recombinant hirudin neither interfered with immunophenotyping of mononuclear cells using FACScan analysis, nor did it alter the detection of Wilms' tumor gene expression by RT-PCR technology even at high doses (5000 ATU hirudin). Thus, a hirudin-containing blood sampling tube can be designed as a universal blood sampling tube (UBT) for testing the majority of diagnostic blood parameters.

  3. Calculation of Blood Dose in Patients Treated With 131I Using MIRD, Imaging, and Blood Sampling Methods.

    PubMed

    Piruzan, Elham; Haghighatafshar, Mahdi; Faghihi, Reza; Entezarmahdi, Seyed Mohammad

    2016-03-01

    Radioiodine therapy is known as the most effective treatment of differentiated thyroid carcinoma (DTC) to ablate remnant thyroid tissue after surgery. In patients with DTC treated with radioiodine, internal radiation dosimetry of radioiodine is useful for radiation risk assessment. The aim of this study is to describe a method to estimate the absorbed dose to the blood using medical internal radiation dosimetry methods. In this study, 23 patients with DTC with different administrated activities, 3.7, 4.62, and 5.55 GBq after thyroidectomy, were randomly selected. Blood dosimetry of treated patients was performed with external whole body counting using a dual-head gamma camera imaging device and also with blood sample activity measurements using a dose calibrator. Absorbed dose to the blood was measured at 2, 6, 12, 24, 48, and 96 hours after the administration of radioiodine with the 2 methods. Based on the results of whole body counting and blood sample activity dose rate measurements, 96 hours after administration of 3.7, 4.62, and 5.55 GBq of radioiodine, absorbed doses to patients' blood were 0.65 ± 0.20, 0.67 ± 0.18, 0.79 ± 0.51 Gy, respectively. Increasing radioiodine activity from 3.7 to 5.55 GBq increased blood dose significantly, while there was no significant difference in blood dose between radioiodine dosages of 3.7 and 4.62 GBq. Our results revealed a significant correlation between the blood absorbed dose and blood sample activity and between the blood absorbed dose and whole body counts 24 to 48 hours after the administration of radioiodine.

  4. Whole blood is the sample matrix of choice for monitoring systemic triclocarban levels.

    PubMed

    Schebb, Nils Helge; Ahn, Ki Chang; Dong, Hua; Gee, Shirley J; Hammock, Bruce D

    2012-05-01

    The antibacterial triclocarban (TCC) concentrates in the cellular fraction of blood. Consequently, plasma levels are at least two-fold lower than the TCC amount present in blood. Utilizing whole blood sampling, a low but significant absorption of TCC from soap during showering is demonstrated for a small group of human subjects.

  5. A femoral arteriovenous shunt facilitates arterial whole blood sampling in animals.

    PubMed

    Weber, Bruno; Burger, Cyrill; Biro, Peter; Buck, Alfred

    2002-03-01

    In this study we evaluated on-line continuous blood sampling in a femoral arteriovenous (a-v) shunt for use in quantitative tracer studies using gamma-emitting radionuclides in animals. The shunt consisted of 40 cm polyethylene tubing (PE-50) guided through a coincidence probe. Two three-way valves allowed blood pressure measurements and tracer injection. Blood flow in the shunt and the impulse response function (IRF) were assessed using heparinized human blood mixed with fluorine-18 fluorodeoxyglucose (FDG). In vivo experiments were performed in eight male rats (300-350 g) anaesthetized with halothane. In three rats, manual blood sampling was performed in parallel with on-line sampling. In another five animals, the arterial whole blood activity was recorded on-line for 40 min. For the experiments 150-180 MBq FDG was injected over 35 s. Blood flow in the shunt was 23.6, 29.2 and 42.8 ml/h at 100, 120 and 160 mmHg, respectively. The IRF was characterized by minimal dispersion (1-2 s FWHM). Deconvolution of the measured arterial input curves with the IRF changed the measured curve only minimally. Whole blood radioactivity concentration derived from manual and on-line sampling were in excellent agreement. The curves derived from on-line sampling were of high statistical quality. In conclusion, a femoral a-v shunt allows multiple manipulations such as measurement of the arterial whole blood activity, continuous blood pressure monitoring, injection of the tracer and collection of blood samples if necessary. It is not associated with blood loss if the collection of blood samples is not required. It is more convenient to use than manual sampling, the peak of the input curve is never missed and the input curves are of high statistical quality.

  6. Novel blood sampling method of an artificial endocrine pancreas via the cardiopulmonary bypass circuit.

    PubMed

    Kawahito, Shinji; Higuchi, Seiichi; Mita, Naoji; Kitagawa, Tetsuya; Kitahata, Hiroshi

    2013-12-01

    We tried to perform continuous blood glucose monitoring during cardiovascular surgery involving cardiopulmonary bypass using an artificial endocrine pancreas (STG-22 or -55; Nikkiso, Tokyo, Japan); however, we often encountered problems during these procedures because insufficient blood was obtained for monitoring. Thus, we started performing the blood sampling via the venous side of the cardiopulmonary bypass circuit. As a result, continuous blood glucose monitoring using an artificial endocrine pancreas was proven to be stable and reliable during cardiovascular surgery involving cardiopulmonary bypass.

  7. Quantitative photoacoustic blood oxygenation measurement of whole porcine blood samples using a multi-wavelength semiconductor laser system

    NASA Astrophysics Data System (ADS)

    Friedrich, Claus-Stefan; Mienkina, Martin P.; Brenner, Carsten; Gerhardt, Nils C.; Jörger, Manfred; Strauß, Andreas; Beckmann, Martin F.; Schmitz, Georg; Hofmann, Martin R.

    2011-07-01

    We present a photoacoustic measurement system based on semiconductor lasers for blood oxygenation measurements. It permits to use four different optical wavelengths (650nm, 808nm, 850nm, 905nm) to generate photoacoustic signals. As the optical extinction coefficient of oxygenated hemoglobin and deoxygenated hemoglobin is different at specific wavelengths, a blood oxygenation measurement by a multi-wavelength photoacoustic laser system is feasible. Especially at 650nm, the clear difference between the extinction coefficients of the two hemoglobin derivates permits to determine the blood oxygenation in combination with other near infrared wavelengths. A linear model based on tabulated values of extinction coefficients for fully oxygenated and fully deoxygenated hemoglobin is presented. We used heparin stabilized whole porcine blood samples to model the optical behavior of human blood, as the optical absorption behavior of porcine hemoglobin does not differ significantly from human hemoglobin. To determine the real oxygen saturation values of the blood samples, we measured the partial oxygen pressure with an IRMA Trupoint Blood Analysis System. The oxygen saturation values were calculated from a dissociation curve for porcine blood. The results of the photoacoustic measurement are in qualitatively good agreement with the predicted linear model. Further, we analyze the abilities and the limitations of quantitative oxygenation measurements.

  8. Sampling errors in pH and blood gas analysis--an evaluation of three new arterial blood samplers.

    PubMed

    Hutchison, A S; Dryburgh, F J; Ralston, S H

    1986-05-01

    We have tested the accuracy, acceptability and general performance of three recently-marketed samplers for arterial blood gas measurement (the Corning Arterial Blood Sampler, the Concord 'Pulsator' and the Sarstedt 'Monovette'). All three greatly reduce or eliminate the error of venous sampling, and the Corning and Sarstedt samplers eliminate the risk of dilution of the sample by excess heparin solution. A positive bias in pO2 measurement, more marked at higher levels, was demonstrated with the Concord and Sarstedt samplers, and the latter carry a slightly increased risk of cross-infection. None of the samplers completely overcame potential sampling errors.

  9. Nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia.

    PubMed

    Krleza, Jasna Lenicek

    2014-01-01

    Capillary sampling is increasingly used to obtain blood for laboratory tests in volumes as small as necessary and as non-invasively as possible. Whether capillary blood sampling is also frequent in Croatia, and whether it is performed according to international laboratory standards is unclear. All medical laboratories that participate in the Croatian National External Quality Assessment Program (N = 204) were surveyed on-line to collect information about the laboratory's parent institution, patient population, types and frequencies of laboratory tests based on capillary blood samples, choice of reference intervals, and policies and procedures specifically related to capillary sampling. Sampling practices were compared with guidelines from the Clinical and Laboratory Standards Institute (CLSI) and the World Health Organization (WHO). Of the 204 laboratories surveyed, 174 (85%) responded with complete questionnaires. Among the 174 respondents, 155 (89%) reported that they routinely perform capillary sampling, which is carried out by laboratory staff in 118 laboratories (76%). Nearly half of respondent laboratories (48%) do not have a written protocol including order of draw for multiple sampling. A single puncture site is used to provide capillary blood for up to two samples at 43% of laboratories that occasionally or regularly perform such sampling. Most respondents (88%) never perform arterialisation prior to capillary blood sampling. Capillary blood sampling is highly prevalent in Croatia across different types of clinical facilities and patient populations. Capillary sampling procedures are not standardised in the country, and the rate of laboratory compliance with CLSI and WHO guidelines is low.

  10. Hematological assessment in pet guinea pigs (Cavia porcellus): blood sample collection and blood cell identification.

    PubMed

    Zimmerman, Kurt; Moore, David M; Smith, Stephen A

    2015-01-01

    Pet guinea pigs are presented to veterinary clinics for routine care and treatment of clinical diseases. In addition to obtaining clinical history and exam findings, diagnostic testing may be required, including hematological assessments. This article describes common blood collection methods, including venipuncture sites, the volume of blood that can be safely collected, and handling of the blood. Hematological parameters for normal guinea pigs are provided for comparison with in-house or commercial test results. A description of the morphology of guinea pig leukocytes is provided to assist in performing a differential count.

  11. Hematological assessment in pet rabbits: blood sample collection and blood cell identification.

    PubMed

    Moore, David M; Zimmerman, Kurt; Smith, Stephen A

    2015-01-01

    Pet rabbits are presented to veterinary clinics for routine care and treatment of clinical diseases. In addition to obtaining clinical history, additional diagnostic testing may be required, including hematological assessments. This article describes common blood collection methods, including venipuncture sites, volume of blood that can be safely collected, and handling of the blood. Hematological parameters for normal rabbits are provided for comparison with in-house or commercial test results. A description of the morphology of rabbit leukocytes is provided to assist in performing a differential count. Differential diagnoses are provided for abnormal values identified in the hemogram.

  12. Hematological Assessment in Pet Guinea Pigs (Cavia porcellus): Blood Sample Collection and Blood Cell Identification.

    PubMed

    Zimmerman, Kurt; Moore, David M; Smith, Stephen A

    2015-09-01

    Pet guinea pigs are presented to veterinary clinics for routine care and treatment of clinical diseases. In addition to obtaining clinical history and exam findings, diagnostic testing may be required, including hematological assessments. This article describes common blood collection methods, including venipuncture sites, the volume of blood that can be safely collected, and handling of the blood. Hematological parameters for normal guinea pigs are provided for comparison with in-house or commercial test results. A description of the morphology of guinea pig leukocytes is provided to assist in performing a differential count.

  13. Hematologic Assessment in Pet Rats, Mice, Hamsters, and Gerbils: Blood Sample Collection and Blood Cell Identification.

    PubMed

    Lindstrom, Nicole M; Moore, David M; Zimmerman, Kurt; Smith, Stephen A

    2015-09-01

    Hamsters, gerbils, rats, and mice are presented to veterinary clinics and hospitals for prophylactic care and treatment of clinical signs of disease. Physical examination, history, and husbandry practice information can be supplemented greatly by assessment of hematologic parameters. As a resource for veterinarians and their technicians, this article describes the methods for collection of blood, identification of blood cells, and interpretation of the hemogram in mice, rats, gerbils, and hamsters.

  14. Hematologic assessment in pet rats, mice, hamsters, and gerbils: blood sample collection and blood cell identification.

    PubMed

    Lindstrom, Nicole M; Moore, David M; Zimmerman, Kurt; Smith, Stephen A

    2015-01-01

    Hamsters, gerbils, rats, and mice are presented to veterinary clinics and hospitals for prophylactic care and treatment of clinical signs of disease. Physical examination, history, and husbandry practice information can be supplemented greatly by assessment of hematologic parameters. As a resource for veterinarians and their technicians, this article describes the methods for collection of blood, identification of blood cells, and interpretation of the hemogram in mice, rats, gerbils, and hamsters.

  15. Effect of red blood cell aggregation and sedimentation on optical coherence tomography signals from blood samples

    NASA Astrophysics Data System (ADS)

    Kirillin, M. Yu; Priezzhev, A. V.; Tuchin, V. V.; Wang, R. K.; Myllylä, R.

    2005-08-01

    In this work, Monte Carlo simulation is used to obtain model optical coherence tomography (OCT) signals from a horizontally orientated blood layer at different stages of red blood cell (RBC) aggregation and sedimentation processes. The parameters for aggregating and sedimenting blood cells were chosen based on the data available from the literature and our earlier experimental studies. We consider two different cases: a suspension of washed RBCs in physiological solution (where aggregation does not take place) and RBCs in blood plasma (which provides necessary conditions for aggregation). Good agreement of the simulation results with the available experimental data shows that the chosen optical parameters are reasonable. The dependence of the numbers of photons contributing to the OCT signal on the number of experienced scattering events was analysed for each simulated signal. It was shown that the maxima of these dependences correspond to the peaks in the OCT signals related to the interfaces between the layers of blood plasma and blood cells. Their positions can be calculated from the optical thicknesses of the layers, and the absorption and scattering coefficients of the media.

  16. A field-deployable device for the rapid detection of cyanide poisoning in whole blood

    NASA Astrophysics Data System (ADS)

    Boehringer, Hans; Tong, Winnie; Chung, Roy; Boss, Gerry; O'Farrell, Brendan

    2012-06-01

    Feasibility of a field-deployable device for the rapid and early diagnosis of cyanide poisoning in whole blood using the spectral shift of the vitamin B12 precursor cobinamide upon binding with cyanide as an indicator is being assessed. Cyanide is an extremely potent and rapid acting poison with as little as 50 mg fatal to humans. Cyanide poisoning has been recognized as a threat from smoke inhalation and potentially through weapons of mass destruction. Currently, no portable rapid tests for the detection of cyanide in whole blood are available. Cobinamide has an extremely high affinity for cyanide and captures hemoglobin associated cyanide from red blood cells. Upon binding of cyanide, cobinamide undergoes a spectral shift that can be measured with a spectrophotometer. We have combined the unique cyanide-binding properties of cobinamide with blood separation technology, sample transport and a detection system, and are developing a rapid, field deployable, disposable device which will deliver an intuitive result to a first responder, allowing for rapid response to exposure events. Feasibility of the cobinamide-Cyanide chemistry in a rapid test using a whole blood sample from a finger-stick has been demonstrated with an assay time from sample collection to a valid result of under 5 minutes. Data showing the efficacy of the diagnostic method and initial device design concepts will be shown.

  17. Effect of blood sampling schedule on the ability to discriminate between postprandial glycemic responses.

    PubMed

    Lui, Janice L; Lan-Pidhainy, Xiaomiao; Brummer, Yolanda; Tosh, Susan M; Wood, Peter J; Wolever, Thomas M S

    2009-10-01

    The blood glucose responses elicited by foods are often determined using blood samples taken at 15-min intervals. Our objective was to see whether taking blood samples at 10-min intervals affected the results. Overnight-fasted healthy subjects (n=11) were studied on nine different occasions with seven different test meals. Blood samples were obtained at fasting and at 10, 15, 20, 30, 40, 45, 50, 60, 90, and 120 min after starting to eat. Peak rise, incremental area under the curve, and relative glycemic response were calculated using the 10- and 15-min sampling schedules. With 10-min intervals, peak rise was 4% greater than with 15-min intervals (P<0.001), but sampling interval did not significantly affect mean incremental area under the curve or relative glycemic response. The 10-min blood sampling schedule had a slightly greater ability to discriminate between foods and between subjects for peak rise and relative glycemic response. We conclude that the blood sampling schedule used may influence the accuracy and precision of measurements of glycemic response; however, the difference between taking blood samples at 10-min and 15-min intervals is quite small.

  18. Glycosylated haemoglobin: comparison of five different methods, including measurement on capillary blood samples.

    PubMed

    Moore, J C; Bown, E; Outlaw, M C; Jelfs, R; Holman, R R; Turner, R C

    1986-01-01

    Glycosylated haemoglobin was measured in venous blood samples and in blood collected in 'Unistep' bottles by isoelectric focusing (IEF), as the reference method, and by electroendosmosis (EEO), the thiobarbituric acid method (TBA), ion-exchange chromatography (IEC) and affinity chromatography (AC). Isoelectric focusing, electroendosmosis and thiobarbituric acid gave similar results. Affinity chromatography gave lower results than isoelectric focusing for normal values but similar results for diabetics. Ion-exchange chromatography gave 24% lower results than isoelectric focusing across the range. Using Unistep collected blood samples and comparing multiple samples from the same patient, electroendosmosis gave the best results (coefficient of variation 4%) and thiobarbituric acid gave slightly less good precision that other methods. Re-use of affinity chromatography columns gave less good precision. Collection of blood samples into a Unistep bottle gave similar results to venous sample results. Storage of venous capillary blood samples in Unistep bottles over 1 week at 21 degrees C gave similar results to immediate assay. Electroendosmosis of blood samples in Unistep bottles gave stable results over 2 weeks. Home collection by a patient of a capillary blood sample into a Unistep bottle allows glycosylated haemoglobin results to be available when seen in the clinic.

  19. Lipid emulsion solution: A novel cause of hemolysis in serum and plasma blood samples.

    PubMed

    Jaben, Elizabeth A; Koch, Christopher D; Karon, Brad S

    2011-02-01

    After several hemolyzed blood samples were received in the laboratory, we investigated lipid emulsion/TPN as a novel cause of hemolysis. Whole blood was spiked with lipid emulsion and TPN. Hemolysis was proportional to the amount of lipid emulsion present in whole blood, with less hemolysis occurring in blood gas syringes compared to vacutainer tubes. Collection of specimens in blood gas syringes may prevent hemolysis in patients on lipid emulsion. Copyright © 2010 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  20. Use of filter paper blood samples for rabies antibody detection in foxes and raccoon dogs.

    PubMed

    Wasniewski, Marine; Barrat, Jacques; Combes, Benoit; Guiot, Anne Laure; Cliquet, Florence

    2014-08-01

    The effectiveness of oral rabies vaccination in wildlife is usually evaluated by the detection of rabies antibodies. However, the assessment of rabies antibodies has several technical difficulties in the field, such as the collection, storage, transport and titration of blood samples, often of poor quality. The objective of this study was to assess the feasibility of collecting blood on a filter paper (FP) coupled with enzyme-linked immunosorbent assay (ELISA) titration of rabies antibodies in raccoon dogs and red foxes. The FP blood sampling method was found highly specific and repeatable in both species. Overall, results obtained with the FP sampling method were highly concordant with the conventional (venipuncture) sampling methods. Blood eluates from FP samples from foxes and raccoon dogs tested using ELISA showed concordance values of 92% and 95%, respectively, with serum samples tested using the seroneutralisation test and values of 95% and 91%, respectively, when the ELISA was used on both types of sample. The use of FP blood sampling coupled with the titration of rabies antibodies by ELISA provides a reliable alternative to conventional blood sampling and serum testing by seroneutralisation. This simple procedure is particularly attractive and cost-effective for assessing the effectiveness of oral rabies vaccination in field conditions.

  1. Impact of blood sample collection and processing methods on glucose levels in community outreach studies.

    PubMed

    Turchiano, Michael; Nguyen, Cuong; Fierman, Arthur; Lifshitz, Mark; Convit, Antonio

    2013-01-01

    Glucose obtained from unprocessed blood samples can decrease by 5%-7% per hour due to glycolysis. This study compared the impact of glucose degradation on measured glucose values by examining two different collection methods. For the first method, blood samples were collected in tubes containing sodium fluoride (NaF), a glycolysis inhibitor. For the second method, blood samples were collected in tubes containing a clot activator and serum gel separator and were centrifuged to separate the serum and plasma 20 minutes after sample collection. The samples used in the two methods were collected during the same blood draw and were assayed by the clinical laboratory 2-4 hours after the samples were obtained. A total of 256 pairs of samples were analyzed. The average glucose reading for the centrifuged tubes was significantly higher than the NaF tubes by 0.196 ± 0.159 mmol/L (P < 0.01) or 4.2%. This study demonstrates the important role collection methods play in accurately assessing glucose levels of blood samples collected in the field, where working environment may be suboptimal. Therefore, blood samples collected in the field should be promptly centrifuged before being transported to clinical labs to ensure accurate glucose level measurements.

  2. DNA methylome profiling using neonatal dried blood spot samples: a proof-of-principle study.

    PubMed

    Hollegaard, Mads Vilhelm; Grauholm, Jonas; Nørgaard-Pedersen, Bent; Hougaard, David Michael

    2013-04-01

    DNA methylation is the most common DNA modification and perhaps the best described epigenetic modification. It is believed to be important for genomic imprinting and gene regulation and has been associated with the development of diseases such as schizophrenia and some types of cancer. Neonatal dried blood spot samples, commonly known as Guthrie cards, are routinely collected worldwide to screen newborns for diseases. Some countries, including Denmark, have been storing the excess neonatal dried blood spot samples in biobanks for decades. Representing a high percentage of the population under a certain age, the neonatal dried blood spot samples are a potential alternative to collecting new samples to study diseases. As such, neonatal dried blood spot samples have previously been used for DNA genotyping studies with excellent results. However, the amount of material available for research is often limited, challenging researchers to generate the most data from a limited quantity of material. In this proof-of-principle study, we address whether two 3.2mm disks punched from a neonatal dried blood spot sample contain enough DNA for genome-wide methylome profiling, measuring 27,578 loci at the same time. We selected two subjects and carried out the following with each: 1) collected an adult whole-blood sample as reference, 2) spotted a fraction of the whole-blood sample onto a similar type of filter paper as used in the newborn screening and stored it for 3years to serve as a dried blood spot reference, and 3) identified the archived neonatal dried blood spot samples, stored for 26-28years, in the Danish Newborn Screening Biobank as a representative of the archived samples. For comparison, we used two different kits for DNA extraction. The DNA, extracted using the Extract-N-Amp Blood PCR kit, was analyzed, and no statistically significant differences were observed (P<0.001) when we compared the methylation profile of the reference whole-blood samples to the dried blood

  3. Algorithm-based arterial blood sampling recognition increasing safety in point-of-care diagnostics.

    PubMed

    Peter, Jörg; Klingert, Wilfried; Klingert, Kathrin; Thiel, Karolin; Wulff, Daniel; Königsrainer, Alfred; Rosenstiel, Wolfgang; Schenk, Martin

    2017-08-04

    To detect blood withdrawal for patients with arterial blood pressure monitoring to increase patient safety and provide better sample dating. Blood pressure information obtained from a patient monitor was fed as a real-time data stream to an experimental medical framework. This framework was connected to an analytical application which observes changes in systolic, diastolic and mean pressure to determine anomalies in the continuous data stream. Detection was based on an increased mean blood pressure caused by the closing of the withdrawal three-way tap and an absence of systolic and diastolic measurements during this manipulation. For evaluation of the proposed algorithm, measured data from animal studies in healthy pigs were used. Using this novel approach for processing real-time measurement data of arterial pressure monitoring, the exact time of blood withdrawal could be successfully detected retrospectively and in real-time. The algorithm was able to detect 422 of 434 (97%) blood withdrawals for blood gas analysis in the retrospective analysis of 7 study trials. Additionally, 64 sampling events for other procedures like laboratory and activated clotting time analyses were detected. The proposed algorithm achieved a sensitivity of 0.97, a precision of 0.96 and an F1 score of 0.97. Arterial blood pressure monitoring data can be used to perform an accurate identification of individual blood samplings in order to reduce sample mix-ups and thereby increase patient safety.

  4. Algorithm-based arterial blood sampling recognition increasing safety in point-of-care diagnostics

    PubMed Central

    Peter, Jörg; Klingert, Wilfried; Klingert, Kathrin; Thiel, Karolin; Wulff, Daniel; Königsrainer, Alfred; Rosenstiel, Wolfgang; Schenk, Martin

    2017-01-01

    AIM To detect blood withdrawal for patients with arterial blood pressure monitoring to increase patient safety and provide better sample dating. METHODS Blood pressure information obtained from a patient monitor was fed as a real-time data stream to an experimental medical framework. This framework was connected to an analytical application which observes changes in systolic, diastolic and mean pressure to determine anomalies in the continuous data stream. Detection was based on an increased mean blood pressure caused by the closing of the withdrawal three-way tap and an absence of systolic and diastolic measurements during this manipulation. For evaluation of the proposed algorithm, measured data from animal studies in healthy pigs were used. RESULTS Using this novel approach for processing real-time measurement data of arterial pressure monitoring, the exact time of blood withdrawal could be successfully detected retrospectively and in real-time. The algorithm was able to detect 422 of 434 (97%) blood withdrawals for blood gas analysis in the retrospective analysis of 7 study trials. Additionally, 64 sampling events for other procedures like laboratory and activated clotting time analyses were detected. The proposed algorithm achieved a sensitivity of 0.97, a precision of 0.96 and an F1 score of 0.97. CONCLUSION Arterial blood pressure monitoring data can be used to perform an accurate identification of individual blood samplings in order to reduce sample mix-ups and thereby increase patient safety. PMID:28828302

  5. Effects of sample storage time, temperature and syringe type on blood gas tensions in samples with high oxygen partial pressures.

    PubMed Central

    Pretto, J. J.; Rochford, P. D.

    1994-01-01

    BACKGROUND--Although plastic arterial sampling syringes are now commonly used, the effects of sample storage time and temperature on blood gas tensions are poorly described for samples with a high oxygen partial pressure (PaO2) taken with these high density polypropylene syringes. METHODS--Two ml samples of tonometered whole blood (PaO2 86.7 kPa, PaCO2 4.27 kPa) were placed in glass syringes and in three brands of plastic blood gas syringes. The syringes were placed either at room temperature or in iced water and blood gas analysis was performed at baseline and after 5, 10, 20, 40, 60, 90, and 120 minutes. RESULTS--In the first 10 minutes measured PaO2 in plastic syringes at room temperature fell by an average of 1.21 kPa/min; placing the sample on ice reduced the rate of PaO2 decline to 0.19 kPa/min. The rate of fall of PaO2 in glass at room temperature was 0.49 kPa/min. The changes in PaCO2 were less dramatic and at room temperature averaged increases of 0.47 kPa for plastic syringes and 0.71 kPa for glass syringes over the entire two hour period. These changes in gas tension for plastic syringes would lead to an overestimation of pulmonary shunt measured by the 100% oxygen technique of 0.6% for each minute left at room temperature before analysis. CONCLUSIONS--Glass syringes are superior to plastic syringes in preserving samples with a high PaO2, and prompt and adequate cooling of such samples is essential for accurate blood gas analysis. PMID:8016801

  6. Comparison of blood chemistry values for samples collected from juvenile chinook salmon by three methods

    USGS Publications Warehouse

    Congleton, J.L.; LaVoie, W.J.

    2001-01-01

    Thirteen blood chemistry indices were compared for samples collected by three commonly used methods: caudal transection, heart puncture, and caudal vessel puncture. Apparent biases in blood chemistry values for samples obtained by caudal transection were consistent with dilution with tissue fluids: alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), creatine kinase (CK), triglyceride, and K+ were increased and Na+ and Cl- were decreased relative to values for samples obtained by caudal vessel puncture. Some enzyme activities (ALT, AST, LDH) and K+ concentrations were also greater in samples taken by heart puncture than in samples taken by caudal vessel puncture. Of the methods tested, caudal vessel puncture had the least effect on blood chemistry values and should be preferred for blood chemistry studies on juvenile salmonids.

  7. Comparison of three methods of sampling trout blood for measurements of hematocrit

    USGS Publications Warehouse

    Steucke, Erwin W.; Schoettger, Richard A.

    1967-01-01

    Trout blood is frequently collected for hematocrit measurements by excising the caudal fin (Snieszko, 1960), but this technique is impractical if valuable fish are to be sampled or if repeated observations are desired. Schiffman (1959) and Snieszko (1960) collected blood from the dorsal aorta and the heart, but these methods are relatively slow and require the preparation of needles and syringes. The use of pointed capillary tubes for cardiac punctures increases the speed of sampling, but body fluids may dilute the blood (Perkins, 1957; Larsen and Snieszko, 1961; and Normandau, 1962). There is need for methods of sampling which are rapid and which neither influence hematological determinations nor harm the fish.

  8. Long-term preservation of blood samples for diagnosis of Trypanosoma cruzi infection.

    PubMed

    Pérez, A C; Cura, E; Subías, E; Lansetti, J C; Segura, E L

    1990-03-01

    Feasability and suitability for field research of a whole-blood preservation method was evaluated through the screening of anti-Trypanosoma cruzi antibodies in 1209 samples under different conditions. Antibody reactivity of paired samples from preserved capillary blood (CBP) and sera from venous blood (VBS) were studied by specific techniques. Over 96% concordance was found on indoor studies carried out with samples without storage or after 15 or 30 days preservation of CBP at 37 degrees C and VBS at -20 degrees C. Outdoor studies performed at field conditions, achieved a 92.1% concordance.

  9. Microwave Blood Thawing: Biochemical Analysis of Small Samples of Thawed Red Blood Cells.

    DTIC Science & Technology

    1984-01-01

    lactate + NAD+ ( Lehninger , 1977) The large increase in pyruvate observed at 6 hours post-wash was most likely due to the large lactate concentrations at...Storage of Blood. London: Academic Press. Lehninger , A.L. 1977. Biochemistry. New York: Worth Publishers, Inc. Lewis, G.P. 1965. Method using o-tolidine

  10. Characterization at the individual cell level and in whole blood samples of shear stress preventing red blood cells aggregation.

    PubMed

    Lee, K; Kinnunen, M; Danilina, A V; Ustinov, V D; Shin, S; Meglinski, I; Priezzhev, A V

    2016-05-03

    The aggregation of red blood cells (RBC) is an intrinsic feature of blood that has a strong impact on its microcirculation. For a number of years it has been attracting a great attention in basic research and clinical studies. Here, we study a relationship between the RBC aggregation parameters measured at the individual cell level and in a whole blood sample. The home made optical tweezers were used to measure the aggregating and disaggregating forces for a pair of interacting RBCs, at the individual cell level, in order to evaluate the corresponding shear stresses. The RheoScan aggregometer was used for the measurements of critical shear stress (CSS) in whole blood samples. The correlation between CSS and the shear stress required to stop an RBC pair from aggregating was found. The shear stress required to disaggregate a pair of RBCs using the double channel optical tweezers appeared to be about 10 times higher than CSS. The correlation between shear stresses required to prevent RBCs from aggregation at the individual cell level and in whole blood samples was estimated and assessed quantitatively. The experimental approach developed has a high potential for advancing hemorheological studies.

  11. Evaluation of a new handheld point-of-care blood gas analyser using 100 equine blood samples.

    PubMed

    Bardell, David; West, Eleanor; Senior, J Mark

    2016-07-07

    To determine whether the Enterprise point-of-care blood analysis system (EPOC) produces results in agreement with two other blood gas analysers in regular clinical use (i-STAT and Radiometer ABL77) and to investigate the precision of the new machine when used with equine whole blood. Prospective, randomized, non-blinded, comparative laboratory analyser study. Horses admitted to a university teaching hospital requiring arterial or venous blood gas analysis as part of their routine clinical management. One hundred equine blood samples were run immediately, consecutively and in randomized order on three blood gas analysers. Results of variables common to all three analysers were tested for agreement and compared with guidelines used in human medicine. These require 80% of results from the test analyser to fall within a defined range or percentage of results from the comparator devices to achieve acceptability. Additionally, 21 samples were run twice in quick succession on the EPOC analyser to investigate precision. Agreement targets were not met for haematocrit, haemoglobin and base excess for either i-STAT or ABL77 analysers. EPOC precision targets were not met for partial pressure of carbon dioxide, ionized calcium, haematocrit and haemoglobin. Overall comparative performance of the EPOC was good to excellent for pH, oxygen tension, potassium, bicarbonate and oxygen saturation of haemoglobin, but marginal to poor for other parameters. The EPOC may be useful in performing analysis of equine whole blood, but trend analysis of carbon dioxide tension, ionized calcium, haematocrit and haemoglobin should be interpreted with caution. The EPOC should not be used interchangeably with other blood gas analysers. © 2016 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia.

  12. Evaluation of a new handheld point-of-care blood gas analyser using 100 equine blood samples.

    PubMed

    Bardell, David; West, Eleanor; Mark Senior, J

    2017-02-22

    To determine whether the Enterprise point-of-care blood analysis system (EPOC) produces results in agreement with two other blood gas analysers in regular clinical use (i-STAT and Radiometer ABL77) and to investigate the precision of the new machine when used with equine whole blood. Prospective, randomized, non-blinded, comparative laboratory analyser study. Horses admitted to a university teaching hospital requiring arterial or venous blood gas analysis as part of their routine clinical management. One hundred equine blood samples were run immediately, consecutively and in randomized order on three blood gas analysers. Results of variables common to all three analysers were tested for agreement and compared with guidelines used in human medicine. These require 80% of results from the test analyser to fall within a defined range or percentage of results from the comparator devices to achieve acceptability. Additionally, 21 samples were run twice in quick succession on the EPOC analyser to investigate precision. Agreement targets were not met for haematocrit, haemoglobin and base excess for either i-STAT or ABL77 analysers. EPOC precision targets were not met for partial pressure of carbon dioxide, ionized calcium, haematocrit and haemoglobin. Overall comparative performance of the EPOC was good to excellent for pH, oxygen tension, potassium, bicarbonate and oxygen saturation of haemoglobin, but marginal to poor for other parameters. The EPOC may be useful in performing analysis of equine whole blood, but trend analysis of carbon dioxide tension, ionized calcium, haematocrit and haemoglobin should be interpreted with caution. The EPOC should not be used interchangeably with other blood gas analysers. Copyright © 2016 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.

  13. The Effect of Pneumatic Tube Systems on the Hemolysis of Biochemistry Blood Samples.

    PubMed

    Cakirca, Gokhan; Erdal, Huseyin

    2017-05-01

    Pneumatic tube systems (PTSs) are widely used in many hospitals because they lead to reduced turnaround times and cost efficiency. However, PTSs may affect the quality of the blood samples transported to the laboratory. The aim of this study was to investigate the effect of the PTS used in our hospital on the hemolysis of the biochemical blood samples transported to the laboratory. A total of 148 samples were manually transported to the laboratory by hospital staff, 148 samples were transported with the PTS, and 113 were transported with the PTS without use of sponge-rubber inserts (PTSws). Hemolysis rates and the levels of biochemical analytes for the different transportation methods were compared. No significant difference was found between the samples transported manually and with the PTS with regard to hemolysis rate and the levels of biochemical analytes. However, the samples transported with the PTSws showed a significant difference compared with the samples transported manually and with the PTS with regard to hemolysis rate and potassium and lactate dehydrogenase levels. The percentages of the samples that exceeded the permissible threshold for the hemolysis among the samples transported manually, with the PTS, and with the PTSws were 10%, 8%, and 47%, respectively. A PTS can be used safely for transporting biochemistry blood samples to the laboratory. However, a sponge-rubber insert that holds sample tubes must be used with the PTS to prevent the hemolysis of blood samples. Copyright © 2016 Emergency Nurses Association. Published by Elsevier Inc. All rights reserved.

  14. Blood cells in rainbow trout Oncorhynchus mykiss milt: relation to milt collection method and sampling period.

    PubMed

    Ciereszko, A; Wlasow, T; Dobosz, S; Goryczko, K; Glogowski, J

    2004-10-01

    The presence of blood cells in milt of rainbow trout (Oncorhynchus mykiss) collected every week between the middle at the end of the spawning season, either by stripping or by catheterization was investigated. Basic sperm biological and biochemical characteristics were also evaluated. Because milt often becomes contaminated with blood during collection, we also studied the influence of experimental blood contamination on sperm motility and biochemical parameters of seminal plasma. We demonstrated the presence of blood cells (erythrocytes, lymphoid, and phagocytes) in rainbow trout milt collected by both methods. Both sampling period and collection method influenced sperm characteristics, however the relationship between these characteristics and blood cells are not clear at present. A high number of blood cells in milt was found in some samples, possibly due to inflammation, because at the same time we observed bacteria and elevated levels of protein and antiproteinase activity in contaminated samples. Experimental contamination of milt with blood did not influence sperm motility, protein concentration and LDH activity of the 5-day-stored semen. Our study demonstrated that blood cells were present in rainbow trout milt. Blood cells may also appear in milt as a result of bleeding and their elevated levels are present during inflammation.

  15. The effects of storage temperature and duration of blood samples on DNA and RNA qualities.

    PubMed

    Huang, Lien-Hung; Lin, Pei-Hsien; Tsai, Kuo-Wang; Wang, Liang-Jen; Huang, Ying-Hsien; Kuo, Ho-Chang; Li, Sung-Chou

    2017-01-01

    DNA and RNA samples from blood are the common examination target for non-invasive physical tests and/or biomedical studies. Since high-quality DNA and RNA samples guarantee the correctness of these tests and/or studies, we investigated the effects of storage temperature and storage duration of whole blood on DNA and RNA qualities. Subjects were enrolled to donate blood samples which were stored for different durations and at different temperatures, followed by the examinations on RNA quality, qPCR, DNA quality and DNA methylation. For RNA, we observed obvious quality decline with storage duration longer than 24 hours. Storage at low temperature does not keep RNA samples from degradation. And, storing whole blood samples in freezer dramatically damage RNA. For DNA, quality decline was not observed even with storage duration for 15 days. However, DNA methylation significantly altered with storage duration longer than three days. Storage duration within 24 hours is critical for collecting high-quality RNA samples for next-generation sequencing (NGS) assays (RIN≧8). If microarray assays are expected (RIN≧7), storage duration within 32 hours is acceptable. Although DNA is resistant within 15 days when kept in whole blood, DNA quantity dramatically decreases owing to WBC lysis. In addition, duration for more than three days significantly alter DNA methylation status, globally and locally. Our result provides a reference for dealing with blood samples.

  16. A nonlethal sampling method to obtain, generate and assemble whole blood transcriptomes from small, wild mammals.

    PubMed

    Huang, Zixia; Gallot, Aurore; Lao, Nga T; Puechmaille, Sébastien J; Foley, Nicole M; Jebb, David; Bekaert, Michaël; Teeling, Emma C

    2016-01-01

    The acquisition of tissue samples from wild populations is a constant challenge in conservation biology, especially for endangered species and protected species where nonlethal sampling is the only option. Whole blood has been suggested as a nonlethal sample type that contains a high percentage of bodywide and genomewide transcripts and therefore can be used to assess the transcriptional status of an individual, and to infer a high percentage of the genome. However, only limited quantities of blood can be nonlethally sampled from small species and it is not known if enough genetic material is contained in only a few drops of blood, which represents the upper limit of sample collection for some small species. In this study, we developed a nonlethal sampling method, the laboratory protocols and a bioinformatic pipeline to sequence and assemble the whole blood transcriptome, using Illumina RNA-Seq, from wild greater mouse-eared bats (Myotis myotis). For optimal results, both ribosomal and globin RNAs must be removed before library construction. Treatment of DNase is recommended but not required enabling the use of smaller amounts of starting RNA. A large proportion of protein-coding genes (61%) in the genome were expressed in the blood transcriptome, comparable to brain (65%), kidney (63%) and liver (58%) transcriptomes, and up to 99% of the mitogenome (excluding D-loop) was recovered in the RNA-Seq data. In conclusion, this nonlethal blood sampling method provides an opportunity for a genomewide transcriptomic study of small, endangered or critically protected species, without sacrificing any individuals.

  17. Quality standards in Biobanking: authentication by genetic profiling of blood spots from donor's original sample

    PubMed Central

    Cardoso, Sergio; Valverde, Laura; Odriozola, Adrian; Elcoroaristizabal, Xabier; de Pancorbo, Marian M

    2010-01-01

    The field of Biobanking requires extensive work to maintain traceability of samples. However, sometimes the necessity to authenticate a sample may arise. To address these circumstances, we herein present a method for authenticating derivatives by using a blood spot from each donor, attached to a sample authentication form, by means of genetic profiling. Blood spots are collected at the time a blood sample is donated at a health centre and before processing the blood sample at the biobank. To test the validity of our approach over time, we analyzed 26 blood spots stored at room temperature in our facilities for more than 15 years. DNA was successfully extracted from the three storage materials tested in this study and 15 STR markers plus amelogenin were subsequently analyzed. The storage of a small blood spot attached to a sample authentication form proved to be efficient for genetic profiling and, therefore, may constitute a long-lasting (at least 15 years), cost-effective and effortless approach for genetic authentication of samples in biobanks. PMID:20234395

  18. Automated Blood Sample Preparation Unit (ABSPU) for Portable Microfluidic Flow Cytometry.

    PubMed

    Chaturvedi, Akhil; Gorthi, Sai Siva

    2017-02-01

    Portable microfluidic diagnostic devices, including flow cytometers, are being developed for point-of-care settings, especially in conjunction with inexpensive imaging devices such as mobile phone cameras. However, two pervasive drawbacks of these have been the lack of automated sample preparation processes and cells settling out of sample suspensions, leading to inaccurate results. We report an automated blood sample preparation unit (ABSPU) to prevent blood samples from settling in a reservoir during loading of samples in flow cytometers. This apparatus automates the preanalytical steps of dilution and staining of blood cells prior to microfluidic loading. It employs an assembly with a miniature vibration motor to drive turbulence in a sample reservoir. To validate performance of this system, we present experimental evidence demonstrating prevention of blood cell settling, cell integrity, and staining of cells prior to flow cytometric analysis. This setup is further integrated with a microfluidic imaging flow cytometer to investigate cell count variability. With no need for prior sample preparation, a drop of whole blood can be directly introduced to the setup without premixing with buffers manually. Our results show that integration of this assembly with microfluidic analysis provides a competent automation tool for low-cost point-of-care blood-based diagnostics.

  19. Accuracy of cyclosporin measurements made in capillary blood samples obtained by skin puncture.

    PubMed

    Merton, G; Jones, K; Lee, M; Johnston, A; Holt, D W

    2000-10-01

    International consensus guidelines suggest that cyclosporin should be measured in whole blood. In some instances it may be advantageous to collect capillary blood, by a finger or ear prick method. However, drug concentrations in skin-puncture blood may not necessarily correlate with those measured in venous blood. This study compared cyclosporin concentrations in blood collected from the fingertip or earlobe with blood collected by standard venipuncture. Patient preference for each of the blood collection methods was also assessed. Specimens were obtained from organ transplant patients receiving cyclosporin, using each of the three methods: venipuncture, finger prick, and earlobe prick. The samples were assayed using a specific radioimmunoassay and the results were compared. In the 102 sets of samples collected, the mean difference (+/- standard deviation) in cyclosporin concentration between finger prick and venipuncture and ear prick and venipuncture was 2.6% (+/- 9.5%) and 2.7% (+/- 12.1%), respectively, while the comparable median (IQR) differences were 1.9% (-3.4% to +6.6%) and -1.1% (-2.8% to +7.2%), respectively. A high degree of correlation was observed between finger prick and venipuncture or ear prick and venipuncture or ear prick and finger prick (r2 > 0.86). Of the three methods of blood collection, finger prick was the patients' preferred method (P < 0.01). These data suggest that capillary blood collected by skin puncture is suitable for use in cyclosporin blood monitoring and acceptable to patients.

  20. Umbilical Arterial Blood Sampling Alters Cerebral Tissue Oxygenation in Very Low Birth Weight Neonates.

    PubMed

    Mintzer, Jonathan P; Parvez, Boriana; La Gamma, Edmund F

    2015-11-01

    To evaluate the magnitude, consistency, and natural history of reductions in cerebral regional tissue oxygenation (CrSO2) during umbilical arterial (UA) blood sampling in very low birth weight neonates. Data were collected during a prospective observational near-infrared spectroscopy survey conducted on a convenience sample of 500-1250 g neonates during the first 10 postnatal days. A before-after analysis of UA blood sampling effects on CrSO2 absolute values and variability was performed. The present analysis was not designed a priori and was conducted following the bedside observation of CrSO2 decrements contiguous with UA blood draws. Fifteen very low birth weight neonates had 201 UA blood draws. Baseline CrSO2 (mean ± SEM) decreased following UA blood sampling, from 70 ± 1% to a nadir of 63 ± 1% (P < .001) occurring 4 ± 3 (range 2-24) minutes following blood draws. CrSO2 subsequently increased to 70 ± 1% (P < .001 compared with nadir) at 10 ± 4 (range 4-28) minutes following UA blood sampling. Coefficients of variation (mean ± SEM) increased from 0.02 ± 0.001 at baseline to 0.05 ± 0.004 (P < .001), followed by a decrease to 0.03 ± 0.003 (P < .001 for all comparisons), thus denoting increased CrSO2 variability following UA blood sampling. UA blood sampling is associated with significant CrSO2 decrements with increased variability over clinically significant intervals. Whether these changes impact complications of prematurity, including intraventricular hemorrhage and periventricular leukomalacia, remain unknown. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Device and method for automated separation of a sample of whole blood into aliquots

    DOEpatents

    Burtis, Carl A.; Johnson, Wayne F.

    1989-01-01

    A device and a method for automated processing and separation of an unmeasured sample of whole blood into multiple aliquots of plasma. Capillaries are radially oriented on a rotor, with the rotor defining a sample chamber, transfer channels, overflow chamber, overflow channel, vent channel, cell chambers, and processing chambers. A sample of whole blood is placed in the sample chamber, and when the rotor is rotated, the blood moves outward through the transfer channels to the processing chambers where the blood is centrifugally separated into a solid cellular component and a liquid plasma component. When the rotor speed is decreased, the plasma component backfills the capillaries resulting in uniform aliquots of plasma which may be used for subsequent analytical procedures.

  2. How You Can Help Medical Research: Donating Your Blood, Tissue, and Other Samples

    MedlinePlus

    National Cancer Institute How You Can Help Medical Research U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES National Institutes of Health Donating Your Blood, Tissue, and Other Samples You have the choice to ...

  3. Influence of an intervention targeting a reduction in sugary beverage intake on the δ13C sugar intake biomarker in a predominantly obese, health-disparate sample.

    PubMed

    Davy, Brenda M; Jahren, A Hope; Hedrick, Valisa E; You, Wen; Zoellner, Jamie M

    2017-01-01

    Controversy exists surrounding the health effects of added sugar (AS) and sugar-sweetened beverage (SSB) intakes, primarily due to a reliance on self-reported dietary intake. The purpose of the current investigation was to determine if a 6-month intervention targeting reduced SSB intake would impact δ13C AS intake biomarker values. A randomized controlled intervention trial. At baseline and at 6 months, participants underwent assessments of anthropometrics and dietary intake. Fasting fingerstick blood samples were obtained and analysed for δ13C value using natural abundance stable isotope MS. Statistical analysis included descriptive statistics, correlational analyses and multilevel mixed-effects linear regression analysis using an intention-to-treat approach. Rural Southwest Virginia, USA. Adults aged ≥18 years who consumed ≥200 kcal SSB/d (≥837 kJ/d) were randomly assigned to either the intervention (n 155) or a matched-contact group (n 146). Participants (mean age 42·1 (sd 13·4) years) were primarily female and overweight (21·5 %) or obese (57·0 %). A significant group by time difference in δ13C value was detected (P<0·001), with mean (sd) δ13C value decreasing in the intervention group (pre: -18·92 (0·65) ‰, post: -18·97 (0·65) ‰) and no change in the comparison group (pre: -18·94 (0·72) ‰, post: -18·92 (0·73) ‰). Significant group differences in weight and BMI change were also detected. Changes in biomarker δ13C values were consistent with changes in self-reported AS and SSB intakes. The δ13C sugar intake biomarker assessed using fingerstick blood samples shows promise as an objective indicator of AS and SSB intakes which could be feasibly included in community-based research trials.

  4. A method for estimating radioactive cesium concentrations in cattle blood using urine samples.

    PubMed

    Sato, Itaru; Yamagishi, Ryoma; Sasaki, Jun; Satoh, Hiroshi; Miura, Kiyoshi; Kikuchi, Kaoru; Otani, Kumiko; Okada, Keiji

    2017-08-04

    In the region contaminated by the Fukushima nuclear accident, radioactive contamination of live cattle should be checked before slaughter. In this study, we establish a precise method for estimating radioactive cesium concentrations in cattle blood using urine samples. Blood and urine samples were collected from a total of 71 cattle on two farms in the 'difficult-to-return zone'. Urine (137) Cs, specific gravity, electrical conductivity, pH, sodium, potassium, calcium, and creatinine were measured and various estimation methods for blood (137) Cs were tested. The average error rate of the estimation was 54.2% without correction. Correcting for urine creatinine, specific gravity, electrical conductivity, or potassium improved the precision of the estimation. Correcting for specific gravity using the following formula gave the most precise estimate (average error rate = 16.9%): [blood (137) Cs] = [urinary (137) Cs]/([specific gravity] - 1)/329. Urine samples are faster to measure than blood samples because urine can be obtained in larger quantities and has a higher (137) Cs concentration than blood. These advantages of urine and the estimation precision demonstrated in our study, indicate that estimation of blood (137) Cs using urine samples is a practical means of monitoring radioactive contamination in live cattle. © 2017 Japanese Society of Animal Science.

  5. Platelet-rich fibrin prepared from stored whole-blood samples.

    PubMed

    Isobe, Kazushige; Suzuki, Masashi; Watanabe, Taisuke; Kitamura, Yutaka; Suzuki, Taiji; Kawabata, Hideo; Nakamura, Masayuki; Okudera, Toshimitsu; Okudera, Hajime; Uematsu, Kohya; Nakata, Koh; Tanaka, Takaaki; Kawase, Tomoyuki

    2017-12-01

    In regenerative therapy, self-clotted platelet concentrates, such as platelet-rich fibrin (PRF), are generally prepared on-site and are immediately used for treatment. If blood samples or prepared clots can be preserved for several days, their clinical applicability will expand. Here, we prepared PRF from stored whole-blood samples and examined their characteristics. Blood samples were collected from non-smoking, healthy male donors (aged 27-67 years, N = 6), and PRF clots were prepared immediately or after storage for 1-2 days. Fibrin fiber was examined by scanning electron microscopy. Bioactivity was evaluated by means of a bioassay system involving human periosteal cells, whereas PDGF-BB concentrations were determined by an enzyme-linked immunosorbent assay. Addition of optimal amounts of a 10% CaCl2 solution restored the coagulative ability of whole-blood samples that contained an anticoagulant (acid citrate dextrose) and were stored for up to 2 days at ambient temperature. In PRF clots prepared from the stored whole-blood samples, the thickness and cross-links of fibrin fibers were almost identical to those of freshly prepared PRF clots. PDGF-BB concentrations in the PRF extract were significantly lower in stored whole-blood samples than in fresh samples; however, both extracts had similar stimulatory effects on periosteal-cell proliferation. Quality of PRF clots prepared from stored whole-blood samples is not reduced significantly and can be ensured for use in regenerative therapy. Therefore, the proposed method enables a more flexible treatment schedule and choice of a more suitable platelet concentrate immediately before treatment, not after blood collection.

  6. Characteristics of a new automated blood sampling system for positron emission tomography

    SciTech Connect

    Eriksson, L.; Ingvar, M.; Rosenqvist, G.; Ekdahl, T.; Kappel, P.

    1995-08-01

    A new commercially available automated blood sampling system (ABSS) for positron emission tomography has been evaluated. The system uses a single BGO crystal and detects with high efficiency the annihilation radiation from tracers, labelled with positron emitting isotopes, in arterial blood. In addition the possibilities to use the ABSS as a detector in the analysis of the plasma samples with liquid chromatography techniques under flow conditions has been explored.

  7. Nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia

    PubMed Central

    Krleza, Jasna Lenicek

    2014-01-01

    Introduction: Capillary sampling is increasingly used to obtain blood for laboratory tests in volumes as small as necessary and as non-invasively as possible. Whether capillary blood sampling is also frequent in Croatia, and whether it is performed according to international laboratory standards is unclear. Materials and methods: All medical laboratories that participate in the Croatian National External Quality Assessment Program (N = 204) were surveyed on-line to collect information about the laboratory’s parent institution, patient population, types and frequencies of laboratory tests based on capillary blood samples, choice of reference intervals, and policies and procedures specifically related to capillary sampling. Sampling practices were compared with guidelines from the Clinical and Laboratory Standards Institute (CLSI) and the World Health Organization (WHO). Results: Of the 204 laboratories surveyed, 174 (85%) responded with complete questionnaires. Among the 174 respondents, 155 (89%) reported that they routinely perform capillary sampling, which is carried out by laboratory staff in 118 laboratories (76%). Nearly half of respondent laboratories (48%) do not have a written protocol including order of draw for multiple sampling. A single puncture site is used to provide capillary blood for up to two samples at 43% of laboratories that occasionally or regularly perform such sampling. Most respondents (88%) never perform arterialisation prior to capillary blood sampling. Conclusions: Capillary blood sampling is highly prevalent in Croatia across different types of clinical facilities and patient populations. Capillary sampling procedures are not standardised in the country, and the rate of laboratory compliance with CLSI and WHO guidelines is low. PMID:25351353

  8. Cerebral Oxygenation and Pain of Heel Blood Sampling Using Manual and Automatic Lancets in Premature Infants.

    PubMed

    Hwang, Mi-Jung; Seol, Geun Hee

    2015-01-01

    Heel blood sampling is a common but painful procedure for neonates. Automatic lancets have been shown to be more effective, with reduced pain and tissue damage, than manual lancets, but the effects of lancet type on cortical activation have not yet been compared. The study aimed to compare the effects of manual and automatic lancets on cerebral oxygenation and pain of heel blood sampling in 24 premature infants with respiratory distress syndrome. Effectiveness was measured by assessing numbers of pricks and squeezes and duration of heel blood sampling. Pain responses were measured using the premature infant pain profile score, heart rate, and oxygen saturation (SpO2). Regional cerebral oxygen saturation (rScO2) was measured using near-infrared spectroscopy, and cerebral fractional tissue oxygen extraction was calculated from SpO2 and rScO. Measures of effectiveness were significantly better with automatic than with manual lancing, including fewer heel punctures (P = .009) and squeezes (P < .001) and shorter duration of heel blood sampling (P = .002). rScO2 was significantly higher (P = .013) and cerebral fractional tissue oxygen extraction after puncture significantly lower (P = .040) with automatic lancing. Premature infant pain profile scores during (P = .004) and after (P = .048) puncture were significantly lower in the automatic than in the manual lancet group. Automatic lancets for heel blood sampling in neonates with respiratory distress syndrome significantly reduced pain and enhanced cerebral oxygenation, suggesting that heel blood should be sampled routinely using an automatic lancet.

  9. Gingival crevicular blood for assessment of blood glucose in diabetic patients.

    PubMed

    Parker, R C; Rapley, J W; Isley, W; Spencer, P; Killoy, W J

    1993-07-01

    In this study 50 patients with diabetes mellitus had gingival crevicular blood from periodontal probing collected in small plastic pipettes. The pipettes transferred the crevicular blood to a non-wipe glucose self-monitoring instrument. At the same time, finger-stick capillary blood measurements were analyzed in the same glucose self-monitoring instrument, and venous blood was collected for measurement in a laboratory glucose analyzer. Each laboratory measurement was corrected from a serum glucose value to a whole blood glucose value by a function of the patient's hematocrit. This corrected glucose value allowed direct comparison of the laboratory measurement to the intraoral and finger-stick whole blood measurements. The patient blood glucose concentrations ranged from 59 mg/dl to 366 mg/dl. The gingival crevicular blood exhibited a correlation of r = 0.975 (P < .0001) to the corrected laboratory standard measurement, with a mean prediction error (bias) of -4.11 mg/dl and a root mean square error (precision) of 17.43 mg/dl. The finger-stick blood had a correlation of r = 0.983 (P < .0001) to the corrected laboratory standard, with a mean prediction error of 4.65 mg/dl and a root mean square error of 14.48 mg/dl. The American Diabetic Association recommends that the prediction error of blood glucose monitoring devices fall within 15% of the laboratory standard. Using this criterion 92% of the gingival crevicular measurements and 90% of the finger-puncture measurements fell within 15% of the laboratory value.(ABSTRACT TRUNCATED AT 250 WORDS)

  10. Study for the improvement of umbilical cord blood sampling using a new trial apparatus.

    PubMed

    Masaoka, Naoki; Morooka, Masako; Nakajima, Yoshiyuki; Ogata, Hajime; Kodo, Hideki; Kato, Shunichi

    2014-02-01

    The aim of this study was to evaluate the usefulness of the trial umbilical cord blood sampling bag for unrelated cord blood transplantation. Data were obtained from 100 vaginal deliveries. In 50 cases, umbilical cord blood (UCB) was taken with the traditional Kawasumi type UCB sampling bag. In another 50 cases, UCB were taken with trial UCB sampling bag offered by NIPRO Co. We compared the sampling volume between the two groups. Furthermore, 10 cases in each group were matched by sampling volume; we examined the quality of UCB on the number and concentration of nucleated cells, mononuclear cells, CD34+ cells and colony-forming unit granulocyte macrophage and the numbers tested positive for bacteria. Whereas there were no significant differences in gestational weeks at sampling, the ratio of primipara women to multipara women, maternal age, and neonatal weight between the two groups, the sampling UCB volumes with the trial sampling bag were significantly higher than those with traditional sampling bags (P < 0.05). In addition, this phenomenon was more significant in the latter part of the study period (P < 0.05). On the other hand, there were no significant differences in the quality of UCB between the two groups. Once clinicians have become accustomed to the trial UBC sampling bag, this method might be a useful method for collecting UCB for unrelated cord blood transplantation. © 2013 The Authors. Journal of Obstetrics and Gynaecology Research © 2013 Japan Society of Obstetrics and Gynecology.

  11. Sample taking during orthopedic surgery: sensitivity and specificity using the BACTEC blood culture system.

    PubMed

    Podleska, L E; Lendemans, S; Schmid, E; Hussmann, B; Nast-Kolb, D; Taeger, G

    2012-02-01

    The use of blood culture systems for sterile body fluids other than blood has proven to be superior to routine culture methods. This study was conducted in order to assess the performance of the BACTEC blood culture system compared to swab/tissue sample collection for the detection of infection from intraoperative samples taken during surgical procedures. Sensitivity was determined by taking samples (BACTEC and swab/tissue samples) from patients with clinically evident infection (Infection group). Specificity was tested by taking the same sample sets from patients who had aseptic operations with no history of infection (Control group). The sensitivity was found to be much higher for the BACTEC group (50 isolates from 56 samples, sensitivity: 89%) compared to the swab/tissue samples (29 isolates out of 56 samples, sensitivity: 52%). The specificity was lower in the BACTEC group (32 isolates out of 44 samples, specificity: 27%) compared to the swab/tissue samples (1 isolate out of 44 samples, specificity: 98%). We conclude that BACTEC is useful for intraoperative sample collection in cases of low-grade infection. However, it is less specific and there is always the possibility for contamination. Therefore, it is advisable to use this technique in combination with regular tissue samples.

  12. Elimination of heparin interference during microarray processing of fresh and biobank-archived blood samples.

    PubMed

    Hebels, Dennie G A J; van Herwijnen, Marcel H M; Brauers, Karen J J; de Kok, Theo M C M; Chalkiadaki, Georgia; Kyrtopoulos, Soterios A; Kleinjans, Jos C S

    2014-07-01

    In the context of environmental health research, biobank blood samples have recently been identified as suitable for high-throughput omics analyses enabling the identification of new biomarkers of exposure and disease. However, blood samples containing the anti-coagulant heparin could complicate transcriptomic analysis because heparin may inhibit RNA polymerase causing inefficient cRNA synthesis and fluorophore labelling. We investigated the inhibitory effect of heparin and the influence of storage conditions (0 or 3 hr bench times, storage at room temperature or -80°C) on fluorophore labelling in heparinized fresh human buffy coat and whole blood biobank samples during the mRNA work-up protocol for microarray analysis. Subsequently, we removed heparin by lithium chloride (LiCl) treatment and performed a quality control analysis of LiCl-treated biobank sample microarrays to prove their suitability for downstream data analysis. Both fresh and biobank samples experienced varying degrees of heparin-induced inhibition of fluorophore labelling, making most samples unusable for microarray analysis. RNA derived from EDTA and citrate blood was not inhibited. No effect of bench time was observed but room temperature storage gave slightly better results. Strong correlations were observed between original blood sample RNA yield and the amount of synthesized cRNA. LiCl treatment restored sample quality to normal standards in both fresh and biobank samples and the previously identified correlations disappeared. Microarrays hybridized with LiCl-treated biobank samples were of excellent quality with no identifiable influence of heparin. We conclude that, to obtain high quality results, in most cases heparin removal is essential in blood-derived RNA samples intended for microarray analysis. Copyright © 2014 Wiley Periodicals, Inc.

  13. Investigation of prothrombin time in human whole-blood samples with a quartz crystal biosensor.

    PubMed

    Müller, Lothar; Sinn, Stefan; Drechsel, Hartmut; Ziegler, Christiane; Wendel, Hans-Peter; Northoff, Hinnak; Gehring, Frank K

    2010-01-15

    Monitoring of blood coagulation and fibrinolysis is an important issue in treatment of patients with cardiovascular problems and in surgery when blood gets into contact with artificial surfaces. In this work a new method for measuring the coagulation time (prothrombin time, PT) of human whole-blood samples based on a quartz crystal microbalance (QCM) biosensor is presented. The 10 MHz sensors used in this work respond with a frequency shift to changes in viscosity during blood clot formation. For driving and for readout of the quartz, both a network analyzer and an oscillator circuit were utilized. The sensor surfaces were specifically coated with a thin polyethylene layer. We found that both frequency analysis methods are suitable to measure exact prothrombin times in a very good conformity with a mechanical coagulometer as a reference. The anticoagulant effect of heparin on the prothrombin time was exemplarily shown as well as the reverse effect of the heparin antagonist polybrene. The change of the viscoelastic properties during blood coagulation, reflected by the ratio of frequency and dissipation shifts, is discussed for different dilutions of the whole-blood samples. In conclusion, QCM is a distinguished biosensor technique to determine prothrombin time and to monitor heparin therapy in whole-blood samples. Due to the excellent potential of miniaturization and the availability of direct digital signals, the method is predestinated for incorporation and integration into other devices and is thus opening the field of application for inline coagulation diagnostic in extracorporeal blood circuits.

  14. Postmortem measurement of caffeine in bone marrow: influence of sample location and correlation with blood concentration.

    PubMed

    Cartiser, N; Bévalot, F; Chatenay, C; Le Meur, C; Gaillard, Y; Malicier, D; Guitton, J; Fanton, L

    2011-07-15

    Bone marrow (BM) analysis is of forensic interest in postmortem toxicological investigation in case of limited, unavailable or unusable blood samples. However, it remains difficult to determine whether a drug BM concentration is therapeutic or represents overdose, due to the lack of studies on this alternative matrix. Given the variations in BM composition in the body, sample location was suggested to be a relevant factor in assessing BM concentration. The aim of the present study was to compare postmortem caffeine concentrations in various BM sample locations and secondly to consider the correlation between BM and blood concentrations. Six BM samples (right and left side: proximal and medial femur and 5th rib) and a blood sample were collected from 21 forensic autopsies. Gas chromatography coupled to tandem mass spectrometry was performed. Blood caffeine concentrations ranged from 60 to 7591ng/mL. Femoral and rib BM concentrations ranged from 51 to 6171ng/g and 66 to 7280ng/g, respectively. Blood concentrations were always higher than BM concentrations. As a good correlation was demonstrated between blood and rib BM and between blood and the average of the four femoral BM concentrations, blood caffeine concentrations could be correctly extrapolated from BM concentrations. BM caffeine concentration was found to depend on sample location. Rib BM caffeine concentrations appeared to be systematically greater than averaged femur values and concentrations were much more variable between the 4 femur BM samples than between the 2 ribs. From a practical point of view, for caffeine analysis, rib BM appeared more relevant than femoral BM, which requires multisampling to overcome the concentration variability problem.

  15. Comparative infectious serology testing of pre- and post-mortem blood samples from cornea donors.

    PubMed

    Wilkemeyer, I; Pruss, A; Kalus, U; Schroeter, J

    2012-08-01

    Defined serological blood tests of deceased cornea donors are required to minimize the risk of viral infections of a transplant recipient as much as possible. Haemolysis, autolysis and bacterial contamination, may produce significant changes of post-mortem blood samples, which may lead to false serological test results. Pre- and post-mortem findings from the same cornea donors of the University Tissue Bank of the Charité in the years 2004-2009 (n = 487) were retrospectively analyzed and compared. The test results from pre-mortem blood samples were defined as the reference for the post-mortem blood test. Of 487 cornea donors, there were a total of 21 cases (4.3%) with discrepancies between serological test results from pre- and post-mortem blood samples. Of these, 7 values referred to the HBsAg-testing, 3 to the anti-HBs-, 1 to the anti-HBcIgG + IgM-, 1 to the anti-HCV-, 4 to the anti-HIV 1/2- and 5 to the TPLA-findings. False negative results within post-mortem serology occurred in 4 of 487 cases (0.8%). False positive results within the post-mortem blood samples occurred at a much more frequent rate, with 17 of 487 cases (3.5%). Discrepancies between serological pre- and post-mortem blood tests occur mainly due to the use of non-validated test systems. Therefore, it seems reasonable to test pre- and post-mortem blood samples serologically, whenever possible, at the same time, regardless of the sample age. Positive results, regardless of the sample type, should always be retested with validated confirmation tests (e.g. NAT), in order to differentiate between false and true positive results.

  16. Hemolysis rates in blood samples: differences between blood collected by clinicians and nurses and the effect of phlebotomy training.

    PubMed

    Cadamuro, Janne; von Meyer, Alexander; Wiedemann, Helmut; Klaus Felder, Thomas; Moser, Franziska; Kipman, Ulrike; Haschke-Becher, Elisabeth; Mrazek, Cornelia; Simundic, Ana-Maria

    2016-12-01

    Hemolytic samples are one of the most challenging preanalytical issues in laboratory medicine. Even causes leading to hemolytic specimen are various, including phlebotomy practices. Respective educational interventions as well as the reduction of the number of people involved in blood collections are claimed to influence the sample quality for the better. In our hospital 70 junior doctors were in charge of routine phlebotomy until 2012, when this task was shifted to 874 nurses, including a preceding training in phlebotomy and preanalytics. Our aim was to evaluate the impact of this training effect and the increase of people involved on sample quality. The hemolysis index (HI) of 43,875 samples was measured before (n=21,512) and after (n=22,363) the switch of blood collection responsibilities. Differences in overall hemolysis rates and the amount of plasma samples with a concentration of free hemoglobin (fHb) above 0.5 g/L and 1 g/L were calculated. Overall HI as well as the percentage of samples with an fHb concentration >0.5 g/L decreased after the responsibility for phlebotomy changed. The rate of samples with an fHb concentration >1 g/L remained unchanged. Hemolysis rates were reduced upon passing phlebotomy tasks from untrained physicians on to a trained nursing staff. We therefore conclude that the number of people performing phlebotomy seems to play a minor role, compared to the effect of a standardized training. However, whether a reduction in the number of people involved in blood collection could lead to further improvement of sample quality, remains to be investigated in future studies.

  17. Effect of delayed processing of blood samples on performance of cytomegalovirus antigenemia assay.

    PubMed Central

    Niubò, J; Pérez, J L; Carvajal, A; Ardanuy, C; Martín, R

    1994-01-01

    This prospective, parallel, and blind study of 120 blood samples from immunocompromised patients examined the influence of delayed sample processing on cytomegalovirus antigenemia assay. Cytomegalovirus was detected in 49 samples (40.8%): 41 were processed within 2 to 4 h, and 40 were reprocessed the following day. Results were discrepant in 17 of the samples with the lowest positive cell counts. Differences between the two days did not reach statistical significance. PMID:8027328

  18. Heat stabilization of blood spot samples for determination of metabolically unstable drug compounds

    PubMed Central

    Blessborn, Daniel; Sköld, Karl; Zeeberg, David; Kaewkhao, Karnrawee; Sköld, Olof; Ahnoff, Martin

    2014-01-01

    Background Sample stability is critical for accurate analysis of drug compounds in biosamples. The use of additives to eradicate the enzymatic activity causing loss of these analytes has its limitations. Results A novel technique for sample stabilization by rapid, high-temperature heating was used. The stability of six commercial drugs in blood and blood spots was investigated under various conditions with or without heat stabilization at 95°C. Oseltamivir, cefotaxime and ribavirin were successfully stabilized by heating whereas significant losses were seen in unheated samples. Amodiaquine was stable with and without heating. Artemether and dihydroartemisinin were found to be very heat sensitive and began to decompose even at 60°C. Conclusion Heat stabilization is a viable technique to maintain analytes in blood spot samples, without the use of chemical additives, by stopping the enzymatic activity that causes sample degradation. PMID:23256470

  19. Limited blood sampling for pharmacokinetic dose tailoring of FVIII in the prophylactic treatment of haemophilia A.

    PubMed

    Björkman, S

    2010-07-01

    The aim of this study was to evaluate the use of limited blood sampling and Bayesian analysis to estimate the pharmacokinetics (PK) and tailor the dose of factor VIII (FVIII) in an individual patient. In a Bayesian analysis, PK parameters are estimated from only a few plasma concentration measurements, using a previously established PK model. First the necessary model was created using intense blood sampling FVIII data from 10 patients. Then FVIII data from another 21 patients were used for 'clinical' evaluation. Three scenarios were created retrospectively by reduction of the original 7-sample data set; blood sampling at 4, 24 and 48 h, at 8 and 30 h and at 24 h after the infusion. PK parameters were estimated for each individual using Bayesian analysis and compared with those obtained using conventional methods from the full data. The accuracy of predictions of FVIII levels during prophylactic treatment 5-17 months later and implications for dose tailoring were also investigated. Blood sampling at 4, 24 and 48 h was found to give practically the same PK information as a full, conventional (7-10-sample) study. Even a single 24-h FVIII level provided adequate data for initial dose tailoring and gave predictions of FVIII levels 5-17 months later that were not appreciably worse than predictions based on the full PK analysis. By contrast, dose tailoring based on body weight failed completely. In conclusion, PK-based dose tailoring of FVIII can be performed using limited blood sampling during prophylactic treatment.

  20. Human T lymphocyte differentiation antigens: effects of blood sample storage on Leu antibody binding

    SciTech Connect

    Hensleigh, P.A.; Waters, V.B.; Herzenberg, L.A.

    1983-05-01

    Current studies of human T lymphocytes and their subsets often use quantitative immunofluorescence analysis with monoclonal antibodies against cell surface antigens. With storage of whole blood or separated mononuclear cells for more than a few hours we have found marked changes in lymphocyte analysis using a fluorescence activated cell sorter (FACS). Experiments were done to determine if these lymphocyte changes were influenced by storage temperature and if lymphocytes could be made more stable by addition of culture media RPMI 1640 to whole blood. Optimal conditions found for blood storage were with with addition of 50% RPMI 1640 and with samples held at room temperature (22 degrees C). With these storage conditions, delay on FACS analysis up to 24 hours did not result in spurious results. When blood samples are collected in places remote from the laboratory or when batch analysis of serially collected samples is desirable, excessive storage times should be avoided.

  1. Detection of Cervical Cancer Analyzing Blood Samples with Raman Spectroscopy and Multivariate Analysis

    NASA Astrophysics Data System (ADS)

    González-Solís, J. L.; Rodríguez-López, J.; Martínez-Espinosa, J. C.; Frausto-Reyes, C.; Jave-Suárez, L. F.; Aguilar-Lemarroy, A. C.; Vargas-Rodríguez, H.; Martínez-Cano, E.

    2010-05-01

    The use of Raman spectroscopy to analyze blood biochemistry and hence distinguish between normal and abnormal blood was investigated. The blood samples were obtained from 20 patients who were clinically diagnosed with cervical cancer and 10 healthy volunteer. The imprint was put under the Olympus microscope and several points were chosen for Raman measurement. All spectra were collected at a Jobin-Yvon LabRAM HR800 Raman Spectrometer with NIR 830 nm laser. It is shown that the serum samples from patients with cervical cancer and from the control group can be discriminated when the multivariate statistical methods of Principal Component Analysis (PCA) and Linear Discriminated Analysis (LDA) is applied to their Raman spectra. The ratios of some band intensities were analyzed and some band ratios were significant and corresponded to proteins, phospholipids, and polysaccharides. The preliminary results suggest that Raman spectroscopy could be a new technique for the detection using just blood samples.

  2. Perfluoroalkyl Acid Concentrations in Blood Samples Subjected to Transportation and Processing Delay.

    PubMed

    Bach, Cathrine Carlsen; Henriksen, Tine Brink; Bossi, Rossana; Bech, Bodil Hammer; Fuglsang, Jens; Olsen, Jørn; Nohr, Ellen Aagaard

    2015-01-01

    In studies of perfluoroalkyl acids, the validity and comparability of measured concentrations may be affected by differences in the handling of biospecimens. We aimed to investigate whether measured plasma levels of perfluoroalkyl acids differed between blood samples subjected to delay and transportation prior to processing and samples with immediate processing and freezing. Pregnant women recruited at Aarhus University Hospital, Denmark, (n = 88) provided paired blood samples. For each pair of samples, one was immediately processed and plasma was frozen, and the other was delayed and transported as whole blood before processing and freezing of plasma (similar to the Danish National Birth Cohort). We measured 12 perfluoroalkyl acids and present results for compounds with more than 50% of samples above the lower limit of quantification. For samples taken in the winter, relative differences between the paired samples ranged between -77 and +38% for individual perfluoroalkyl acids. In most cases concentrations were lower in the delayed and transported samples, e.g. the relative difference was -29% (95% confidence interval -30; -27) for perfluorooctane sulfonate. For perfluorooctanoate there was no difference between the two setups [corresponding estimate 1% (0, 3)]. Differences were negligible in the summer for all compounds. Transport of blood samples and processing delay, similar to conditions applied in some large, population-based studies, may affect measured perfluoroalkyl acid concentrations, mainly when outdoor temperatures are low. Attention to processing conditions is needed in studies of perfluoroalkyl acid exposure in humans.

  3. Perfluoroalkyl Acid Concentrations in Blood Samples Subjected to Transportation and Processing Delay

    PubMed Central

    Bach, Cathrine Carlsen; Henriksen, Tine Brink; Bossi, Rossana; Bech, Bodil Hammer; Fuglsang, Jens; Olsen, Jørn; Nohr, Ellen Aagaard

    2015-01-01

    Background In studies of perfluoroalkyl acids, the validity and comparability of measured concentrations may be affected by differences in the handling of biospecimens. We aimed to investigate whether measured plasma levels of perfluoroalkyl acids differed between blood samples subjected to delay and transportation prior to processing and samples with immediate processing and freezing. Methods Pregnant women recruited at Aarhus University Hospital, Denmark, (n = 88) provided paired blood samples. For each pair of samples, one was immediately processed and plasma was frozen, and the other was delayed and transported as whole blood before processing and freezing of plasma (similar to the Danish National Birth Cohort). We measured 12 perfluoroalkyl acids and present results for compounds with more than 50% of samples above the lower limit of quantification. Results For samples taken in the winter, relative differences between the paired samples ranged between -77 and +38% for individual perfluoroalkyl acids. In most cases concentrations were lower in the delayed and transported samples, e.g. the relative difference was -29% (95% confidence interval -30; -27) for perfluorooctane sulfonate. For perfluorooctanoate there was no difference between the two setups [corresponding estimate 1% (0, 3)]. Differences were negligible in the summer for all compounds. Conclusions Transport of blood samples and processing delay, similar to conditions applied in some large, population-based studies, may affect measured perfluoroalkyl acid concentrations, mainly when outdoor temperatures are low. Attention to processing conditions is needed in studies of perfluoroalkyl acid exposure in humans. PMID:26356420

  4. The prevalence of toxoplasmosis in Imam Reza Hospital blood bank samples, Tehran, Iran.

    PubMed

    Shaddel, M; Mirzaii Dizgah, I; Sharif, F

    2014-10-01

    The prevalence of toxoplasma gondii (T.g) infection in blood donors has been poorly studied. The aim of this study was to assess the prevalence of acute and chronic toxoplasmosis in blood products. A total of 223 blood products (101 fresh frozen plasma (FFP) and 122 packed cells (PC)) in Imam Reza hospital blood bank, Tehran, Iran were tested for specific T.g antibodies (IgG and IgM) by ELISA method. Positive IgG anti-T.g samples were further tested for IgM anti-T.g. A positive IgG test with the negative and positive IgM test was interpreted as a chronic and acute toxoplasmosis respectively. Of 223 samples 38.6% and 0.45% were positive for IgG anti-T.g and IgM anti-T.g levels respectively. Therefore, one and 85 samples were involved acute and chronic toxoplasmosis respectively. Twenty-six of fresh frozen plasma samples were positive for IgG anti-T.g and one of them was positive for IgM anti-T.g. Sixty packed cell samples were positive for IgG anti-T.g. Our study showed that there were chronic and acute toxoplasmosis in blood products and the prevalence of toxoplasmosis especially chronic form was high. Therefore screening of blood for T.g antibodies may be considered. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Liquid chromatography coupled with multi-channel electrochemical detection for the determination of daidzin in rat blood sampled by an automated blood sampling system.

    PubMed

    Tian, Feifei; Zhu, Yongxin; Long, Hong; Cregor, Meloney; Xie, Fuming; Kissinger, Candice B; Kissinger, Peter T

    2002-05-25

    Daidzin, a soy-derived biologically active natural product, has been reported to inhibit mitochondrial aldehyde dehydrogenase and suppress ethanol intake. This paper describes a method for the determination of daidzin in rat blood. After administration of daidzin, blood samples were periodically collected from awake, freely moving animals by a Culex automated blood sampler. Daidzin was extracted from 50 microl of diluted blood (blood and saline at a ratio of 1:1) with ethyl acetate. Chromatographic separation was achieved within 12 min using a microbore C(18) (100 x 1.0 mm) 3 microm column with a mobile phase containing 20 mM sodium acetate, 0.25 mM EDTA, pH 4.3, 4% methanol and 11% acetonitrile at a flow-rate of 90 microl/min. Detection was attained using a four-channel electrochemical detector with glassy carbon electrodes using oxidation potentials of +1100, 950, 850, 750 mV vs. Ag/AgCl. The limit of detection for daidzin in rat plasma was 5 ng/ml at a signal-to-noise ratio of 3:1. The extraction recovery of daidzin from rat plasma was over 74%. Linearity was obtained for the range of 25-1000 ng/ml. The intra- and inter-assay precisions were in the ranges of 2.7-6.6 and 1.9-3.7%, respectively. This method is suitable to routine in vivo monitoring of daidzin in rat plasma.

  6. Smart fast blood counting of trace volumes of body fluids from various mammalian species using a compact custom-built microscope cytometer (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Smith, Zachary J.; Gao, Tingjuan; Lin, Tzu-Yin; Carrade-Holt, Danielle; Lane, Stephen M.; Matthews, Dennis L.; Dwyre, Denis M.; Wachsmann-Hogiu, Sebastian

    2016-03-01

    Cell counting in human body fluids such as blood, urine, and CSF is a critical step in the diagnostic process for many diseases. Current automated methods for cell counting are based on flow cytometry systems. However, these automated methods are bulky, costly, require significant user expertise, and are not well suited to counting cells in fluids other than blood. Therefore, their use is limited to large central laboratories that process enough volume of blood to recoup the significant capital investment these instruments require. We present in this talk a combination of a (1) low-cost microscope system, (2) simple sample preparation method, and (3) fully automated analysis designed for providing cell counts in blood and body fluids. We show results on both humans and companion and farm animals, showing that accurate red cell, white cell, and platelet counts, as well as hemoglobin concentration, can be accurately obtained in blood, as well as a 3-part white cell differential in human samples. We can also accurately count red and white cells in body fluids with a limit of detection ~3 orders of magnitude smaller than current automated instruments. This method uses less than 1 microliter of blood, and less than 5 microliters of body fluids to make its measurements, making it highly compatible with finger-stick style collections, as well as appropriate for small animals such as laboratory mice where larger volume blood collections are dangerous to the animal's health.

  7. Standardization of whole blood assay for determination of pyrogenic activity in organic dust samples.

    PubMed

    Liebers, Verena; Stubel, Heike; Düser, Maria; Brüning, Thomas; Raulf-Heimsoth, Monika

    2009-09-01

    To characterize bioaerosol exposure at workplaces standardized methods are necessary. Activity of endotoxin, one component of organic dust, can be quantified with the Limulus-Amoebocyte Lysat test (LAL test). Further information with respect to pyrogenic activity of the organic dust can be achieved by measuring cytokine release of human blood after stimulation with the dust or its extract (whole blood assay). The aim of our study was the standardization of the whole blood assay (WBA) while using cryo-preserved human blood (Qualis Laboratorium) and to compare the outcome of the different cytokines determined by incubation of the blood cells with extracts from dust samples collected at various workplaces. Cytokine release (IL-1 beta, IL-6, IL-8, TNF-alpha, MCP-1) was measured by ELISA after stimulation of fresh blood from ten donors as well as cryo-preserved human blood. In both cases blood was stimulated with E. coli endotoxin as well as with 30 dust filter extracts from various workplaces. All dust filter extracts were investigated in the WBA using cryo-preserved blood as well as with LAL test. E. coli endotoxin stimulated the release of IL-1 beta, IL-6, IL-8, TNF-alpha and MCP-1 in a dose-dependent manner in fresh as well as cryo-preserved human whole blood. 200 pg/ml E. coli endotoxin induced maximal cytokine release in cryo-preserved blood (mean value for IL-1 beta 2509+/-418 pg/ml; n=13 experiments) whereas fresh blood of single donors reached a maximum release when stimulated with 50 ng/ml endotoxin (mean value of ten donors 1125+/-553 pg/ml IL-1beta). Using cryo-preserved blood the coefficient of variation (CV) regarding the interassay variability was below 21% for all cytokines measured. Regarding 26 dust sample extracts correlation coefficient r2 for LAL test and WBA was between 0.90 and 0.93 (Pearson) for IL-1 beta, IL-6, IL-8 and TNF-alpha whereas correlation for MCP-1 was lower (r(2)=0.59). Two dust sample extracts which showed similar reactivity

  8. Maternal red blood cell alloantibodies identified in blood samples obtained from Iranian pregnant women: the first population study in Iran.

    PubMed

    Shahverdi, Ehsan; Moghaddam, Mostafa; Gorzin, Fateme

    2017-01-01

    The objective was to determine the frequency of occurrence of alloantibodies among pregnant women in Iran. This was a prospective cross-sectional study, which was carried out in the immunohematology reference laboratory of the Iranian Blood Transfusion Organization in Tehran, Iran, in 2008 to 2015. Screening and identification of red blood cell (RBC) alloantibodies was done on the sera of 7340 pregnant females using the standard tube method and gel column agglutination technique. Alloantibodies were identified in the serum of 332 of the 7340 (4.5%) pregnant women. A total of 410 antibodies were detected in 332 positive maternal serum samples with no previous history of blood transfusion. Anti-D was the most common antibody accounting for 70.5% of all the antibodies formed in D- women. The incidence of specific alloimmunization other than Rh group was 14.4%. We concluded that the alloimmunization rate was high in comparison with wide pattern in previous studies. In Iran, like other developing countries, alloimmunization screening tests are performed only to detect anti-D in pregnant D- women. This high rate of alloimmunization, quite possibly, is due to the fact that the majority of blood samples came from pregnant women known to have previous obstetric problems. However, we suggest that RBC antibody screening tests should be extended to all D+ women. © 2016 AABB.

  9. Method of evaluation of process of red blood cell sedimentation based on photometry of droplet samples.

    PubMed

    Aristov, Alexander; Nosova, Ekaterina

    2017-04-01

    The paper focuses on research aimed at creating and testing a new approach to evaluate the processes of aggregation and sedimentation of red blood cells for purpose of its use in clinical laboratory diagnostics. The proposed method is based on photometric analysis of blood sample formed as a sessile drop. The results of clinical approbation of this method are given in the paper. Analysis of the processes occurring in the sample in the form of sessile drop during the process of blood cells sedimentation is described. The results of experimental studies to evaluate the effect of the droplet sample focusing properties on light radiation transmittance are presented. It is shown that this method significantly reduces the sample volume and provides sufficiently high sensitivity to the studied processes.

  10. A point-of-care paper-based fingerstick transaminase test: towards low-cost “lab-on-a-chip” technology for the developing world

    PubMed Central

    Pollock, Nira R.; Colby, Donn; Rolland, Jason P.

    2013-01-01

    There is currently great need for high-quality, low-cost, point-of-care diagnostics that can benefit patients in resource-limited settings, and correspondingly growing interest in the diagnostic utility of microfluidic platforms based on paper. We describe the development, early clinical testing, and potential clinical impact of a novel paper-based, multiplexed microfluidic assay designed for rapid, semi-quantitative measurement of AST and ALT in a fingerstick specimen. This device ultimately holds promise for providing universal access to affordable point-of-care screening for drug-induced liver injury in resource-limited settings, and opens the door to development of similar point-of-care clinical assays for other important analytes. PMID:23466712

  11. [Quantitation of cerebral blood flow and partition coefficient using 123I-IMP dynamic SPECT with single arterial blood sampling].

    PubMed

    Mizumura, S; Kumita, S; Kumazaki, T

    1996-03-01

    A method base on the two-compartment model was developed to measure quantitative cerebral blood flow (CBF) and partition coefficient (lambda) of IMP from dynamic SPECT and single arterial blood sampling. In this method, the linear differential equation of two-compartment model, Yokoi proposed, was employed and quantitative CBF and lambda values were measured with the standard input function calibrated by single arterial sampling. The input function was derived from the standard input function scaled by a factor determined by the single arterial blood sample. This new technique was applied to 5 normal volunteers (Ages ranged from 25 to 29 yr., average 26 yr.). The optimal time to calibrate the standard input function in the individual study and optimal the period of the upper limit time to which input function is integrated from IMP administration for analysis of the equation were determined to minimize the difference between integration of the calibrated standard input function and of the individual input function. Minimization of the difference yields an optimal calibration time (4 to 10 min after IMP administration) and the period of the upper limit time (8 to 60 min after acquisition start). Comparison of CBF and lambda values obtained by the graphical method using the calibrated standard data and individual input function were performed. It should be noted that CBF values were in good agreement between the two methods, respectively (r = 0.92, P<0.01; r = 0.72, p = 0.01). This method is easy to estimate CBF and lambda by only single arterial blood sampling and IMP dynamic SPECT, and useful for routine studies.

  12. Simple cannulation procedure for serial blood sampling through cutaneous ulnar vein in chickens.

    PubMed

    Bayer, Darmel M; Mohan, K; Jayakumar, K; Manafi, Milad; Pavithra, B H

    2012-01-01

    The objective of the study was to collect repeated, low-stress blood samples from the ulnar vein of chickens required for pharmacokinetic studies or hormonal assays. The study used 5 apparently healthy, unsexed, commercial broiler chickens about 6 weeks old and weighing 1.7-1.9 kg for serial sampling of blood. The study prepared the birds prior to cannulation and penetrated the catheter through the skin and into the lumen of the ulnar vein. The study successfully carried out serial blood samplings in 4 of 5 cannulated birds. Heparin (10%) solution maintained patency and prevented blood clot formation inside the cannula. However, the study found repeated clotting occurring in 1 bird. Cannula failed to maintain patency; the study could not carry out blood sampling properly, which was attributed to air embolism that might have occurred during catheter manipulation or repeated filling of cannula with heparin solution. The study observed no hematoma or inflammation at the site of cannulation. Owing to the advantages and to facilitate compliance with nonhuman animal welfare, this technique seems simple and efficient, allowing adoption for serial blood collection in chickens.

  13. Paper membrane-based SERS platform for the determination of glucose in blood samples.

    PubMed

    Torul, Hilal; Çiftçi, Hakan; Çetin, Demet; Suludere, Zekiye; Boyacı, Ismail Hakkı; Tamer, Uğur

    2015-11-01

    In this report, we present a paper membrane-based surface-enhanced Raman scattering (SERS) platform for the determination of blood glucose level using a nitrocellulose membrane as substrate paper, and the microfluidic channel was simply constructed by wax-printing method. The rod-shaped gold nanorod particles were modified with 4-mercaptophenylboronic acid (4-MBA) and 1-decanethiol (1-DT) molecules and used as embedded SERS probe for paper-based microfluidics. The SERS measurement area was simply constructed by dropping gold nanoparticles on nitrocellulose membrane, and the blood sample was dropped on the membrane hydrophilic channel. While the blood cells and proteins were held on nitrocellulose membrane, glucose molecules were moved through the channel toward the SERS measurement area. Scanning electron microscopy (SEM) was used to confirm the effective separation of blood matrix, and total analysis is completed in 5 min. In SERS measurements, the intensity of the band at 1070 cm(-1) which is attributed to B-OH vibration decreased depending on the rise in glucose concentration in the blood sample. The glucose concentration was found to be 5.43 ± 0.51 mM in the reference blood sample by using a calibration equation, and the certified value for glucose was 6.17 ± 0.11 mM. The recovery of the glucose in the reference blood sample was about 88 %. According to these results, the developed paper-based microfluidic SERS platform has been found to be suitable for use for the detection of glucose in blood samples without any pretreatment procedure. We believe that paper-based microfluidic systems may provide a wide field of usage for paper-based applications.

  14. The effectiveness of cooling conditions on temperature of canine EDTA whole blood samples

    PubMed Central

    Sun, Xiaocun; Flatland, Bente

    2016-01-01

    Background Preanalytic factors such as time and temperature can have significant effects on laboratory test results. For example, ammonium concentration will increase 31% in blood samples stored at room temperature for 30 min before centrifugation. To reduce preanalytic error, blood samples may be placed in precooled tubes and chilled on ice or in ice water baths; however, the effectiveness of these modalities in cooling blood samples has not been formally evaluated. The purpose of this study was to evaluate the effectiveness of various cooling modalities on reducing temperature of EDTA whole blood samples. Methods Pooled samples of canine EDTA whole blood were divided into two aliquots. Saline was added to one aliquot to produce a packed cell volume (PCV) of 40% and to the second aliquot to produce a PCV of 20% (simulated anemia). Thirty samples from each aliquot were warmed to 37.7 °C and cooled in 2 ml allotments under one of three conditions: in ice, in ice after transfer to a precooled tube, or in an ice water bath. Temperature of each sample was recorded at one minute intervals for 15 min. Results Within treatment conditions, sample PCV had no significant effect on cooling. Cooling in ice water was significantly faster than cooling in ice only or transferring the sample to a precooled tube and cooling it on ice. Mean temperature of samples cooled in ice water was significantly lower at 15 min than mean temperatures of those cooled in ice, whether or not the tube was precooled. By 4 min, samples cooled in an ice water bath had reached mean temperatures less than 4 °C (refrigeration temperature), while samples cooled in other conditions remained above 4.0 °C for at least 11 min. For samples with a PCV of 40%, precooling the tube had no significant effect on rate of cooling on ice. For samples with a PCV of 20%, transfer to a precooled tube resulted in a significantly faster rate of cooling than direct placement of the warmed tube onto ice. Discussion Canine

  15. The effectiveness of cooling conditions on temperature of canine EDTA whole blood samples.

    PubMed

    Tobias, Karen M; Serrano, Leslie; Sun, Xiaocun; Flatland, Bente

    2016-01-01

    Preanalytic factors such as time and temperature can have significant effects on laboratory test results. For example, ammonium concentration will increase 31% in blood samples stored at room temperature for 30 min before centrifugation. To reduce preanalytic error, blood samples may be placed in precooled tubes and chilled on ice or in ice water baths; however, the effectiveness of these modalities in cooling blood samples has not been formally evaluated. The purpose of this study was to evaluate the effectiveness of various cooling modalities on reducing temperature of EDTA whole blood samples. Pooled samples of canine EDTA whole blood were divided into two aliquots. Saline was added to one aliquot to produce a packed cell volume (PCV) of 40% and to the second aliquot to produce a PCV of 20% (simulated anemia). Thirty samples from each aliquot were warmed to 37.7 °C and cooled in 2 ml allotments under one of three conditions: in ice, in ice after transfer to a precooled tube, or in an ice water bath. Temperature of each sample was recorded at one minute intervals for 15 min. Within treatment conditions, sample PCV had no significant effect on cooling. Cooling in ice water was significantly faster than cooling in ice only or transferring the sample to a precooled tube and cooling it on ice. Mean temperature of samples cooled in ice water was significantly lower at 15 min than mean temperatures of those cooled in ice, whether or not the tube was precooled. By 4 min, samples cooled in an ice water bath had reached mean temperatures less than 4 °C (refrigeration temperature), while samples cooled in other conditions remained above 4.0 °C for at least 11 min. For samples with a PCV of 40%, precooling the tube had no significant effect on rate of cooling on ice. For samples with a PCV of 20%, transfer to a precooled tube resulted in a significantly faster rate of cooling than direct placement of the warmed tube onto ice. Canine EDTA whole blood samples cool most

  16. Lead and cadmium levels in blood samples from the general population of Sweden

    SciTech Connect

    Elinder, C.G.; Friberg, L.; Lind, B.; Jawaid, M.

    1983-02-01

    Lead and cadmium was determined in whole blood samples obtained from 473 nonoccupationally exposed adult persons in Sweden in 1980. Analyses were performed using atomic absorption spectrophotometry equipped with an electrothermal atomization unit. Blood lead concentrations were shown to be significantly influenced by sex, smoking habits, and alcohol consumption. Current male smokers had a median blood lead level of 92 ..mu..g Pb/liter, as compared to 77 ..mu..g Pb/liter for nonsmokers. For females the corresponding values were 69 ..mu..g Pb/liter and 57 ..mu..g Pb/liter for current smokers and nonsmokers, respectively. Highly significant correlations were found between stated alcohol consumption and blood lead in most of the different sex and smoking categories. People living in apartments close to streets with heavy traffic in Stockholm had slightly, but not significantly, higher blood lead levels when compared to people living in areas of this city with low traffic density. Blood cadmium levels were very strongly affected by smoking habits. A significant correlation existed between the number of cigarettes consumed daily and blood cadmium concentration. The median blood cadmium level for nonsmoking males was 0.2 ..mu..g Cd/liter (less than or equal to0.2, detection limit) and for females 0.3 ..mu..g Cd/liter. About 90% of the current male and female smokers had cadmium concentrations in blood of 0.6 ..mu..g Cd/liter or more.

  17. [Investigation of uninterpretative HLA typing in 311 umbilical cord blood samples].

    PubMed

    Hong, Jing-Xin; Liang, Xiao-Lan; Han, Jun-Ling; Li, Qian; Qiu, Lu-Gui

    2009-10-01

    The aim of this study was to investigate the factors which affect HLA typing in 311 umbilical cord blood (UCB) samples. The HLA low resolution typing of UCB samples with misinterpreted HLA types from 311 UCB samples analyzed by PCR-SSO and PCR-SSP was performed. 7 samples difficult to determine their HLA genotype were sequenced directly and the reason leading to misinterpret HLA typing was analyzed. The results indicated that 99.4% of misinterpreted samples resulted from the restriction of HLA typing method itself and 0.6% of misinterpreted samples were suspected to be contaminated with maternal blood in UCB. It is concluded that HLA typing is mainly affected by the shortcomings of oligonucleotide probe design for PCR-SSO and lack of allele specific primers of PCR-SSP.

  18. Portable simultaneous multiple analyte whole-blood analyzer for point-of-care testing.

    PubMed

    Schembri, C T; Ostoich, V; Lingane, P J; Burd, T L; Buhl, S N

    1992-09-01

    We describe a portable clinical chemistry analyzer for point-of-care measurements of multiple analytes in less than 10 min from approximately 40 microL of whole blood (fingerstick or venous). Whole blood is applied directly to a 7.9-cm-diameter, single-use plastic rotor containing liquid diluent and greater than or equal to 4-12 tests in the form of 1- to 2-mm-diameter dry reagent beads. The reagent/rotor is immediately placed in a portable instrument along with a ticket/label results card. As the instrument spins the rotor, capillary and rotational forces process the blood into diluted plasma, distribute the patient's diluted sample to cuvettes containing the reagent beads, and mix the diluted sample with the reagents. The instrument monitors the chemical reactions optically at nine wavelengths; sample volume and temperature are also measured optically. The calibration data for each reagent are read from a bar code on the periphery of each rotor. The instrument processes all the measurements to calculate, store, print, and communicate the results. Each reagent/rotor contains an enzymatic control that must be within a defined range before the results from that analysis are reported.

  19. [Isolation of periodontal bacteria from blood samples and atheromas in patients with atherosclerosis and periodontitis].

    PubMed

    Padilla E, Carlos; Lobos G, Olga; Jure O, Gema; Matus F, Sergio; Descouvieres C, Claudia; Hasbún A, Sandra; Maragaño L, Patricio; Núñez F, Loreto

    2007-09-01

    Periodontitis is a common oral disease produced by bacterial species that reside in the subgingival plaque. These microorganisms have been associated to atherosclerosis and it is suggested that periodontitis is a cardiovascular risk factor. To isolate periodontal bacteria from blood and atheroma samples, from patients with atherosclerosis and periodontitis. Twelve patients with periodontitis and a clinical diagnosis of atherosclerosis and 12 patients with periodontitis but without atherosclerosis were studied. Blood samples were obtained immediately before and after scaling and root planing. The samples were incubated in aerobic and anaerobic conditions. One week after scaling, atheromatous plaques were obtained during endarterectomy in the 12 patients with atherosclerosis. These were homogenized and cultured for aerobic and anaerobic bacteria. Microorganisms were identified by means ofPCR. Five patients with and two without atherosclerosis, had bacteremia after scaling and root planing. Bacterial species isolated from blood samples were the same found in periodontic pockets. Four atheromatous plaques of patients with bacteremia yielded positive cultures. The isolated bacteria were the same found in blood samples and periodontal pockets. Bacteremia occurred in seven of 24 patients after scaling and root planing. In four patients, the same species found in periodontic pockets and blood cultures were detected in atherosclerotic plaques obtained one week after the dental procedure.

  20. Rapid and reliable determination of the halogenating peroxidase activity in blood samples.

    PubMed

    Flemmig, Jörg; Schwarz, Pauline; Bäcker, Ingo; Leichsenring, Anna; Lange, Franziska; Arnhold, Jürgen

    2014-12-15

    By combining easy and fast leukocyte enrichment with aminophenyl-fluorescein (APF) staining we developed a method to quickly and specifically address the halogenating activity of the immunological relevant blood heme peroxidases myeloperoxidase and eosinophil peroxidase, respectively. For leukocyte enrichment a two-fold hypotonic lysis procedure of the blood with Millipore water was chosen which represents a cheap, fast and reliable method to diminish the amount of erythrocytes in the samples. This procedure is shown to be suitable both to human and murine blood micro-samples, making it also applicable to small animal experiments with recurring blood sampling. As all types of leukocytes are kept in the sample during the preparation, they can be analysed separately after discrimination during the flow cytometry analysis. This also holds for all heme peroxidase-containing cells, namely neutrophils, eosinophils and monocytes. Moreover additional parameters (e.g. antibody staining) can be combined with the heme peroxidase activity determination to gain additional information about the different immune cell types. Based on previous results we applied APF for specifically addressing the halogenating activity of leukocyte peroxidases in blood samples. This dye is selectively oxidized by the MPO and EPO halogenation products hypochlorous and hypobromous acid. This approach may provide a suitable tool to gain more insights into the immune-physiological role of the halogenating activity of heme peroxidases.

  1. Enhanced Detection of Rift Valley Fever Virus using Molecular Assays on Whole Blood Samples

    PubMed Central

    Grolla, Allen; Mehedi, Masfique; Lindsay, Robbin; Bosio, Catharine; Duse, Adriano; Feldmann, Heinz

    2012-01-01

    Background Rift Valley fever (RVF) is an emerging arthropod-borne zoonoses of global agricultural and public health importance. In December 2006, an RVF outbreak was recognized in Kenya which led to the deployment of international response laboratory teams to the area. Objectives A field laboratory was operated in Malindi, Kenya to provide safe sample handling and molecular testing for RVF virus (RVFV) as well as selected other pathogens for differential diagnosis. Study Design Safe sample handling was carried out using a negative pressure flexible film isolator (glovebox) and commercial reagents to inactivate clinical specimens and purify nucleic acid. Whole blood was routinely used for diagnostic testing although paired plasma samples were also tested in select cases. Subsequently, human macrophages were tested in vitro for their susceptibility to RVFV. Results The field laboratory received samples from 33 individuals and a definite laboratory diagnosis was provided in 16 of these cases. Using molecular diagnostic techniques, RVFV was more consistently detected in whole blood than in plasma samples most likely due to association of RVFV with blood cells. Subsequent in vitro studies identified macrophages as a target cell for RVFV replication. Conclusions RVFV appears to replicate in blood cells such as macrophages. Thus, the sensitivity of molecular diagnostic testing is improved if whole blood is used as the clinical specimen rather than plasma or serum. PMID:22632901

  2. Impact of complete blood count sampling time change on white blood cell and absolute neutrophil count values in clozapine recipients.

    PubMed

    McKee, Jerry R; Wall, Trossie; Owensby, Jessica

    2011-04-01

    Despite its superior efficacy, clozapine is typically reserved for treatment-refractory schizophrenia due to the risk of agranulocytosis with an occurrence of up to 1% in recipients. The FDA has rigid treatment guidelines for hematologic monitoring for clozapine patients. If the white blood cell (WBC) count or absolute neutrophil count (ANC) falls below predetermined values, clozapine treatment must be held or discontinued. Diurnal and ethnic variations in complete blood count (CBC) values, somewhat dependent upon blood sampling time have been reported, and called pseudoneutropenia, which appears independent of clozapine therapy. Unnecessary treatment interruption or discontinuation is costly and may lead to disease relapse. The purpose of this study was to evaluate the effect of a time change in CBC sampling on WBC and ANC values in a group of clozapine patients in a regional public inpatient psychiatric facility. Facility CBC sampling for clozapine patients was switched from 0630 to on or after 0830. A retrospective record review was conducted for all patients who were receiving clozapine before and after the time switch, with a minimum of six values pre- and post-change. CBC values sampled on or after 0830 were accepted as applicable post data, as patients are awakened daily at 0630, and a minimum of two hours of wakefulness/mobility had occurred. Patient medical records, automated lab information system, and the Clozapine National Registry were data sources. Data extracted included WBC/ANC values (with date/time of sampling) and demographic information (DOB, sex, weight, height, BMI, and ethnicity). The data were analyzed using repeated measures ANOVA. Ten patients (80% male, 90% Caucasian, mean age=55.7 years) met study criteria. The difference in the pre/post time change WBC values was marginally significant (mean increase=667/mm3, p=.07), with a significant difference (mean increase=1,130/mm3, p=.003) between the pre/post time change ANC values. ANC values

  3. Using blood samples to estimate persistent organic pollutants and metals in green sea turtles (Chelonia mydas).

    PubMed

    van de Merwe, Jason P; Hodge, Mary; Olszowy, Henry A; Whittier, Joan M; Lee, Shing Y

    2010-04-01

    Persistent organic pollutants (POPs) and heavy metals have been reported in a number of green turtle (Chelonia mydas) populations worldwide. However, due to ethical considerations, these studies have generally been on tissues from deceased and stranded animals. The purpose of this study was to investigate the use of blood samples to estimate the tissue contamination of live C. mydas populations. This study analysed 125 POP compounds and eight heavy metals in the blood, liver, kidney and muscle of 16 C. mydas from the Sea World Sea Turtle Rehabilitation Program, Gold Coast, Australia. Strong correlations were observed between blood and tissue concentrations for a number of POPs and metals. Furthermore, these correlations were observed over large ranges of turtle size, sex and condition. These results indicate that blood samples are a reliable non-lethal method for predicting chemical contamination in C. mydas.

  4. Fast multi-spectral imaging technique for detection of circulating endothelial cells in human blood samples

    NASA Astrophysics Data System (ADS)

    Berezhnyy, Ihor V.; Berezhna, Svitlana Y.

    2012-08-01

    The appearance of non-blood cells circulating in human peripheral bloodstream indicates an abnormal condition. One important category of these cells is circulating endothelial cells (CECs) shed by compromised blood vessels. Clinical applications that measure the blood level of CECs are hindered due to a lack of standardized instruments. The major challenge in detecting circulating non-blood cells is their extreme scarcity; 1 in 106 to 107. Described here is a new method for detection of rare cells in blood samples deposited on the adhesive microscopic slides and immunostained with distinct fluorescent markers. The key novelty of the proposed approach is an intelligent search principle and a dual-mode scanner to implement this principle. To begin, a fast scanning that uses a single beam is performed in the spectral channel where only rare cells produce florescence. Once a target cell is registered, the scanner switches on the imaging mode, auto-focuses and then records images in multiple spectral channels at the selected area. The instrument runs in repetitive cycles until the entire slide is scanned. The technology has been validated via detection of human umbilical vein endothelial cells spiked into human blood samples. In addition, the operational principle can be adapted for detection of other types of rare cells in blood.

  5. Evaluation of pesticide residues in human blood samples from Punjab (India)

    PubMed Central

    Bedi, Jasbir Singh; Gill, J. P. S.; Kaur, P.; Sharma, A.; Aulakh, R. S.

    2015-01-01

    Aim: The present study was undertaken to estimate the current status of residues of organochlorine pesticides (OCPs), organophosphates (OPs) and synthetic pyrethroids (SPs) pesticides in human blood. Materials and Methods: Human blood samples were analyzed by gas chromatography and confirmed by gas chromatography-mass spectrometry in selective ion monitoring mode. Results: The gas chromatographic analysis of human blood samples collected from Punjab revealed the presence of p,p’-dichlorodiphenyl dichloroethylene (DDE), p,p’ dichlorodiphenyl dichloroethane (DDD), o,p’ DDE and β-endosulfan at mean levels of 15.26, 2.71, 5.62 and 4.02 ng/ml, respectively. p,p’ DDE residue was observed in 18.0% blood samples, and it contributes 55% of the total pesticide burden in human blood. The difference of total dichlorordiphenyl trichloroethane (DDT) between different age groups of humans was found to be statistically significant (p<0.05). The difference of DDT and endosulfan between dietary habits, gender and spraying of pesticides was found statistically non-significant, however endosulfan residues were observed only in pesticide sprayer’s population. Conclusion: Occurrence of p,p’ DDE, p,p’ DDD, o,p’ DDE in human blood indicated restricted use of DDT. However, presence of endosulfan residues in occupationally exposed population is a matter of public health concern. PMID:27046999

  6. Blood sampling through peripheral venous catheters is reliable for selected basic analytes in children.

    PubMed

    Berger-Achituv, Sivan; Budde-Schwartzman, Britta; Ellis, Martin H; Shenkman, Ze'ev; Erez, Ilan

    2010-07-01

    The goal was to determine the interchangeability of peripheral venous catheter (PVC) and venipuncture blood sampling (BS). Paired blood samples from hospitalized children were obtained through venipuncture and from existing PVCs, following discard of 2 mL of blood. Comparisons of 9 complete blood count indices (white and red blood cell counts, hemoglobin and hematocrit levels, mean corpuscular volume, mean corpuscular hemoglobin level, red blood cell distribution width, platelet count, and mean platelet volume) and 5 basic chemical analysis indices (sodium, potassium, glucose, chloride, and urea levels) were performed, and hemolysis was documented. Irrespective of gauge, blood samples were obtained successfully from 40 (85.1%) of 47 PVCs, with no abnormal hemolysis. BS through venipuncture took longer than BS from PVCs (175.8 +/- 229.6 vs 104.5 +/- 53.4 seconds; P = .053) and was associated with significantly more distress/crying (73.1% vs 0%; P < .001). There were no significant differences between venipuncture and PVC samples (paired t test). Twenty-one (6%) of 348 pairs analyzed with the Clinical Laboratory Improvement Amendment standards fell outside the range of acceptable variance (8 of 21 aberrations were attributed to glucose measurements). Bland-Altman analysis indicated that, with the exclusion of glucose measurements, BS from PVCs is reliable, with 29 (6.5%) of 448 pairs exceeding the limits of agreement. Of those, 9 cases were clinically significant, but none would have altered clinical management. PVC sampling was shown to be a pain-reducing method that can be used for children for selected basic analytes. The findings for glucose were unreliable.

  7. Prevalence of blood doping in samples collected from elite track and field athletes.

    PubMed

    Sottas, Pierre-Edouard; Robinson, Neil; Fischetto, Giuseppe; Dollé, Gabriel; Alonso, Juan Manuel; Saugy, Martial

    2011-05-01

    No reliable estimate of the prevalence of doping in elite sports has been published. Since 2001, the international governing body for athletics has implemented a blood-testing program to detect altered hematological profiles in the world's top-level athletes. A total of 7289 blood samples were collected from 2737 athletes out of and during international athletic competitions. Data were collected in parallel on each sample, including the age, sex, nationality, and birth date of the athlete; testing date; sport; venue; and instrument technology. Period prevalence of blood-doping in samples was estimated by comparing empirical cumulative distribution functions of the abnormal blood profile score computed for subpopulations with stratified reference cumulative distribution functions. In addition to an expected difference between endurance and nonendurance athletes, we found nationality to be the major factor of heterogeneity. Estimates of the prevalence of blood doping ranged from 1% to 48% for subpopulations of samples and a mean of 14% for the entire study population. Extreme cases of secondary polycythemia highlighted the health risks associated with blood manipulations. When applied at a population level, in this case the population of samples, hematological data can be used to estimate period prevalence of blood doping in elite sports. We found that the world's top-level athletes are not only heterogeneous in physiological and anthropometric factors but also in their doping behavior, with contrasting attitudes toward doping between countries. When applied at the individual level, the same biomarkers, as formalized in the Athlete Biological Passport paradigm, can be used in analysis of the observed different physiological characteristics and behavioral heterogeneities.

  8. Using a Single Blood Sample and Inulin to Estimate Glomerular Filtration Rate in Rabbits

    PubMed Central

    Michigoshi, Yuuki; Yamagishi, Norio; Satoh, Hiroshi; Kato, Masaki; Furuhama, Kazuhisa

    2011-01-01

    To establish a simple procedure for estimating the glomerular filtration rate (GFR) in conscious rabbits, we used the conventional multisample approach to develop a single-blood-sample method. A bolus injection of inulin was administered intravenously at a dose of 40 mg/kg to male New Zealand White rabbits, and blood was collected 30, 60, 90, and 120 min later. Serum inulin, urea nitrogen, and creatinine concentrations were determined. Using this multi-sample method, the reference GFR in clinically healthy rabbits was 4.01 ± 0.17 mL/min/kg (n = 17). In rabbits given an intravenous injection of the antitumor agent cisplatin, GFR fell before serum urea nitrogen and creatinine concentrations increased. Based on cumulative GFR data from healthy and nephropathy rabbits, the GFR obtained from the 3-sample method (30-, 60-, and 90-min samples) was closely correlated (r = 0.99) with that calculated from the estimated distribution volume and serum inulin concentration at 90 min after inulin injection in the single-blood-sample method. These results demonstrate that the single-blood-sample method supports sequential GFR measurements in rabbits and is a versatile procedure not only for research purposes but also in clinical settings. PMID:22330718

  9. Microwaving Blood as a Non-Destructive Technique for Haemoglobin Measurements on Microlitre Samples

    PubMed Central

    Basey-Fisher, Toby H.; Guerra, Nadia; Triulzi, Chiara; Gregory, Andrew; Hanham, Stephen M.; Stevens, Molly M.; Maier, Stefan A.; Klein, Norbert

    2016-01-01

    The non-destructive ex vivo determination of haemoglobin (Hgb) concentration offers the capability to conduct multiple red blood cell haematological measurements on a single sample, an advantage that current optical techniques are unable to offer. Here, a microwave method and device for the accurate and non-destructive determination of Hgb concentration in microlitre blood samples are described. Using broadband microwave spectroscopy, a relationship is established between the dielectric properties of murine blood and Hgb concentration that is utilized to create a technique for the determination of Hgb concentration. Subsequently, a microwave dielectric resonator-microfluidic system is implemented in the analysis of 52 murine samples with microlitre volumes and Hgb concentrations ranging from 0 to 17 g dL−1. Using the characterized relationship, independent and minimally invasive Hgb measurements are made on nine healthy mice as well as seven with mutations in the Adenomatous polyposis coli (APC) gene that leads to colorectal cancer and consequently anaemia. PMID:24002989

  10. A sample-to-result system for blood coagulation tests on a microfluidic disk analyzer

    PubMed Central

    Lin, Chia-Hui; Liu, Cheng-Yuan; Shih, Chih-Hsin; Lu, Chien-Hsing

    2014-01-01

    In this report, we describe in detail a microfluidic analyzer, which is able to conduct blood coagulation tests using whole blood samples. Sample preparation steps, such as whole blood aliquoting and metering, plasma separation, decanting, and mixing with reagents were performed in sequence through microfluidic functions integrated on a disk. Both prothrombin time (PT) and activated partial thromboplastin time (aPTT) were carried out on the same platform and the test results can be reported in 5 min. Fifty clinical samples were tested for both PT and aPTT utilizing the microfluidic disk analyzer and the instrument used in hospitals. The test results showed good correlation and agreement between the two instruments. PMID:25332733

  11. Pneumatic tube-transported blood samples in lithium heparinate gel separator tubes may be more susceptible to haemolysis than blood samples in serum tubes.

    PubMed

    Böckel-Frohnhöfer, Nicole; Hübner, Ulrich; Hummel, Björn; Geisel, Jürgen

    2014-10-01

    Pneumatic tube systems are widely used in hospitals. Advantages are high speed and rapid availability of the samples. However, the transportation by pneumatic tube promotes haemolysis. Haemolysis interferes with many spectrophotometric assays and is a common problem in clinical laboratories. The haemolysis index (HI) as a semi-quantitative representation of the level of haemolysis was compared in unpaired tube-transported and hand-delivered routine lithium heparinate plasma samples (n = 1368 and n = 837, respectively). Additionally, the HI distribution was measured in lithium heparinate plasma samples with a HI above the threshold value of 20 and in paired serum samples after transportation by pneumatic tube system. HI values above 20 can interfere with the selected assays: Creatine kinase (CK), creatine kinase-MB (CK-MB) and alanine aminotransferase (ALT) activities. These parameters were determined to demonstrate how haemolysis affects the results. 17.5% of the tube-transported plasma samples and 2.6% of the hand-delivered plasma samples had a HI above 20. The median HI in pneumatic tube-transported lithium heparinate plasma was 85 and 33 in the paired serum samples. The median HI difference between paired plasma and serum was 46. Blood samples in lithium heparinate tubes may be substantially more susceptible to haemolysis by pneumatic tube transportation than serum tube samples. Although our results cannot be universally applied to laboratories with different pneumatic tube systems, it is recommended that each laboratory evaluate carefully the degree of haemolysis after the transportation by the own pneumatic tube system and in terms of the sample type.

  12. Identifying the potential of changes to blood sample logistics using simulation.

    PubMed

    Jørgensen, Pelle; Jacobsen, Peter; Poulsen, Jørgen Hjelm

    2013-01-01

    Using simulation as an approach to display and improve internal logistics at hospitals has great potential. This study shows how a simulation model displaying the morning blood-taking round at a Danish public hospital can be developed and utilized with the aim of improving the logistics. The focus of the simulation was to evaluate changes made to the transportation of blood samples between wards and the laboratory. The average- (AWT) and maximum waiting time (MWT) from a blood sample was drawn at the ward until it was received at the laboratory, and the distribution of arrivals of blood samples in the laboratory were used as the evaluation criteria. Four different scenarios were tested and compared with the current approach: (1) Using AGVs (mobile robots), (2) using a pneumatic tube system, (3) using porters that are called upon, or (4) using porters that come to the wards every 45 minutes. Furthermore, each of the scenarios was tested in terms of what amount of resources would give the optimal result. The simulations showed a big improvement potential in implementing a new technology/mean for transporting the blood samples. The pneumatic tube system showed the biggest potential lowering the AWT and MWT with approx. 36% and 18%, respectively. Additionally, all of the scenarios had a more even distribution of arrivals except for porters coming to the wards every 45 min. As a consequence of the results obtained in the study, the hospital decided to implement a pneumatic tube system.

  13. Microfluidic, marker-free isolation of circulating tumor cells from blood samples

    PubMed Central

    Karabacak, Nezihi Murat; Spuhler, Philipp S; Fachin, Fabio; Lim, Eugene J; Pai, Vincent; Ozkumur, Emre; Martel, Joseph M; Kojic, Nikola; Smith, Kyle; Chen, Pin-i; Yang, Jennifer; Hwang, Henry; Morgan, Bailey; Trautwein, Julie; Barber, Thomas A; Stott, Shannon L; Maheswaran, Shyamala; Kapur, Ravi; Haber, Daniel A; Toner, Mehmet

    2014-01-01

    The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen–independent microfluidic CTC-iChip technology. The CTC-iChip uses deterministic lateral displacement, inertial focusing and magnetophoresis to sort up to 107 cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2–5 d. PMID:24577360

  14. Microfluidic, marker-free isolation of circulating tumor cells from blood samples.

    PubMed

    Karabacak, Nezihi Murat; Spuhler, Philipp S; Fachin, Fabio; Lim, Eugene J; Pai, Vincent; Ozkumur, Emre; Martel, Joseph M; Kojic, Nikola; Smith, Kyle; Chen, Pin-i; Yang, Jennifer; Hwang, Henry; Morgan, Bailey; Trautwein, Julie; Barber, Thomas A; Stott, Shannon L; Maheswaran, Shyamala; Kapur, Ravi; Haber, Daniel A; Toner, Mehmet

    2014-03-01

    The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen-independent microfluidic CTC-iChip technology. The CTC-iChip uses deterministic lateral displacement, inertial focusing and magnetophoresis to sort up to 10⁷ cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2-5 d.

  15. Rapid Microbial Sample Preparation from Blood Using a Novel Concentration Device

    PubMed Central

    Boardman, Anna K.; Campbell, Jennifer; Wirz, Holger; Sharon, Andre; Sauer-Budge, Alexis F.

    2015-01-01

    Appropriate care for bacteremic patients is dictated by the amount of time needed for an accurate diagnosis. However, the concentration of microbes in the blood is extremely low in these patients (1–100 CFU/mL), traditionally requiring growth (blood culture) or amplification (e.g., PCR) for detection. Current culture-based methods can take a minimum of two days, while faster methods like PCR require a sample free of inhibitors (i.e., blood components). Though commercial kits exist for the removal of blood from these samples, they typically capture only DNA, thereby necessitating the use of blood culture for antimicrobial testing. Here, we report a novel, scaled-up sample preparation protocol carried out in a new microbial concentration device. The process can efficiently lyse 10 mL of bacteremic blood while maintaining the microorganisms’ viability, giving a 30‑μL final output volume. A suite of six microorganisms (Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, Pseudomonas aeruginosa, and Candida albicans) at a range of clinically relevant concentrations was tested. All of the microorganisms had recoveries greater than 55% at the highest tested concentration of 100 CFU/mL, with three of them having over 70% recovery. At the lowest tested concentration of 3 CFU/mL, two microorganisms had recoveries of ca. 40–50% while the other four gave recoveries greater than 70%. Using a Taqman assay for methicillin-sensitive S. aureus (MSSA)to prove the feasibility of downstream analysis, we show that our microbial pellets are clean enough for PCR amplification. PCR testing of 56 spiked-positive and negative samples gave a specificity of 0.97 and a sensitivity of 0.96, showing that our sample preparation protocol holds great promise for the rapid diagnosis of bacteremia directly from a primary sample. PMID:25675242

  16. Venepuncture is preferable to heel lance for blood sampling in term neonates

    PubMed Central

    Ogawa, S; Ogihara, T; Fujiwara, E; Ito, K; Nakano, M; Nakayama, S; Hachiya, T; Fujimoto, N; Abe, H; Ban, S; Ikeda, E; Tamai, H

    2005-01-01

    Background: The analgesic effect of oral sucrose in newborn infants undergoing painful procedures is generally accepted. For blood sampling, some studies have shown that venepuncture (VP) is less painful than heel lance (HL). Objective: To determine the least painful and most effective method among blood sampling by VP or HL with or without sucrose. Design: Randomised, double blind, placebo controlled trial. Subjects: A total of 100 healthy, full term newborn infants being screened for inborn errors of metabolism were randomly allocated to one of four experimental groups (25 infants in each). Intervention and outcome measure: Seven specially trained nurses took turns to carry out blood sampling two minutes after administration of oral sucrose or water. Neonatal pain was assessed by the neonatal facial coding system (NFCS), as well as by crying. Results: Without sucrose, the NFCS score was higher in the HL group than the VP group during blood sampling (median 58 v 23, p<0.001). Oral sucrose significantly reduced the score of the HL group (58 v 47, p<0.01) and also tended to reduce the score of the VP group (23 v 2, p<0.1). However, the HL with sucrose group still had a higher score than the VP without sucrose group (47 v 23, p<0.01). Crying and the total procedure time showed the same trends as the NFCS score. Conclusions: VP is less painful and more effective than HL for blood sampling in newborn infants. Although oral sucrose may have an additive analgesic effect, it is not necessarily required if VP is used for blood sampling. PMID:15871991

  17. Blood Samples of Peripheral Venous Catheter or The Usual Way: Do Infusion Fluid Alters the Biochemical Test Results?

    PubMed Central

    Taghizadeganzadeh, Mahboobeh; Yazdankhahfard, Mohammadreza; Farzaneh, Mohammadreza; Mirzaei, Kamran

    2016-01-01

    Background: Most blood tests require venous blood samples. Puncturing the vein also causes pain, infection, or damage to the blood, and lymph flow, or long-term healing. This study aimed to determine and compare the biochemical laboratory value of the blood samples that were provided through: peripheral vein infusion (PVI) receiving continuous intravenous fluid; and the usual method of blood sampling. Methods: This is an interventional, quasi-experimental, and controlled study. The selected study sample included 60 patients, who were hospitalized during 2014, in the Internal Medicine, part of Martyrs of Persian Gulf, teaching hospital at Bushehr. Three blood samples were taken from each patient that were provided through PVI line (5 ml blood collected at beginning of IVC and then another 5 cc), and another case was prepared by common blood sampling (control). All the samples were analyzed in terms of sodium, potassium, urea and creatinine using SPSS Ver.19 software, by paired t-test and Pearson’s correlation coefficients. Results: There was a statistically significant difference between the amount of sodium and potassium in the first blood samples taken from the intravenous infusion line and vein puncture. However, no significant differences were found among the biochemical amount in the second blood samples taken from the intravenous infusion line and vein puncture. Conclusions: We can use blood samples taken from peripheral intravenous infusion lines after 5cc discarding from the first part of the sample for measuring the value of sodium, potassium, urea and creatinine. PMID:26925892

  18. Effects of anesthesia and blood sampling techniques on plasma metabolites and corticosterone in the rat.

    PubMed

    Arnold, Myrtha; Langhans, Wolfgang

    2010-04-19

    Blood is routinely sampled from laboratory animals in biomedical research, and many of the commonly applied sampling techniques require anesthesia. Acute effects of many sampling and anesthesia procedures may confound the results, but those effects are incompletely characterized. We here compare the effects of four common anesthesia procedures (inhalation anesthesia with ether (EA) or isoflurane (IA) and intraperitoneal injection anesthesia with xylazin/ketamine (XKA) or medetomidine/midazolam/fentanyl (MMFA)) on plasma concentrations of glucose, lactate, non-esterified fatty acids (NEFAs), and corticosterone in blood obtained from a previously implanted jugular vein (JV) catheter with the effect of JV blood sampling from non-anesthetized, freely-moving rats (JV-NA). Also, we included in the comparison two other blood sampling procedures usually performed without anesthesia (NA), i.e., puncture of the saphenic vein (SV) and tail incision (TI). Whereas the control procedure (JV-NA) did not significantly affect any of the target parameters, plasma glucose increased from 14 (JV-IA) to 44 (JV-MMFA) % (all Ps=0.05 when compared with the control procedure) in all blood samples collected in anesthesia and was 12 and 14% lower (both Ps<0.05) in SV-NA and TI-NA samples, respectively. Plasma lactate increased from 74 (JV-IA) to 226% (SV-NA) (all Ps<0.05) with all sampling and anesthesia procedures except for JV-XKA and JV-MMF. Plasma NEFAs increased to 52% (P<0.05) with the TI-NA procedure and appeared to decrease with the JV-IA and JV-MMFA procedures (both Ps>0.05). Finally, only the JV-EA and the JV-MMFA procedures increased plasma corticosterone (+525 and +353%, respectively, both Ps< 0.05). The JV-IA and JV-XKA procedures appeared to increase it as well, but these differences did not reach statistical significance. Thus, anesthesia and blood sampling procedures can have profound acute effects on plasma metabolite and hormone concentrations. This must be considered for

  19. Techniques for Nonterminal Blood Sampling in Black-Tailed Prairie Dogs (Cynomys ludovicianus).

    PubMed

    Head, Valerie; Eshar, David; Nau, Melissa R

    2017-03-01

    Black-tailed prairie dogs (Cynomys ludovicianus) are used as an animal model for research on gallbladder stones and several infectious diseases. A comprehensive, instructive resource regarding the appropriate techniques for venipuncture and collection of nonterminal blood samples in this species has not yet been published. Blood samples (1 mL or larger) were readily obtained from the jugular vein, femoral vein, or cranial vena cava, whereas peripheral sites, such as the cephalic vein, saphenous vein, and tarsal vein, mainly were useful for obtaining smaller volumes. The detailed and illustrated information presented here can aid clinicians and researchers in performing venipuncture, anesthesia, and handling of this species.

  20. Techniques for Nonterminal Blood Sampling in Black-Tailed Prairie Dogs (Cynomys ludovicianus)

    PubMed Central

    Head, Valerie; Eshar, David; Nau, Melissa R

    2017-01-01

    Black-tailed prairie dogs (Cynomys ludovicianus) are used as an animal model for research on gallbladder stones and several infectious diseases. A comprehensive, instructive resource regarding the appropriate techniques for venipuncture and collection of nonterminal blood samples in this species has not yet been published. Blood samples (1 mL or larger) were readily obtained from the jugular vein, femoral vein, or cranial vena cava, whereas peripheral sites, such as the cephalic vein, saphenous vein, and tarsal vein, mainly were useful for obtaining smaller volumes. The detailed and illustrated information presented here can aid clinicians and researchers in performing venipuncture, anesthesia, and handling of this species. PMID:28315653

  1. Techniques for Nonterminal Blood Sampling in Black-Tailed Prairie Dogs (Cynomys ludovicianus).

    PubMed

    Head, Valerie; Eshar, David; Nau, Melissa

    2017-02-13

    Black-tailed prairie dogs (Cynomys ludovicianus) are used as an animal model for research on gallbladder stones and several infectious diseases. A comprehensive, instructive resource regarding the appropriate techniques for venipuncture and collection of nonterminal blood samples in this species has not yet been published. Blood samples (1 mL or larger) were readily obtained from the jugular vein, femoral vein, or cranial vena cava, whereas peripheral sites, such as the cephalic vein, saphenous vein, and tarsal vein, mainly were useful for obtaining smaller volumes. The detailed and illustrated informationpresented here can aid clinicians and researchers in performing venipuncture, anesthesia, and handling of this species.

  2. A Novel Needle-Free Blood Draw Device for Sample Collection From Short Peripheral Catheters

    PubMed Central

    Nachamkin, Irving

    2017-01-01

    A new US Food and Drug Administration-cleared needleless blood collection device (PIVO; Velano Vascular, San Francisco, CA) for short peripheral catheters was compared with conventional venipuncture for collecting blood samples for routine laboratory analysis from adult healthy volunteers. The PIVO device was comparable with venipuncture in terms of providing high-integrity samples (no hemolysis or clotting), equivalent laboratory values, and better patient experience as assessed by pain scores. Further studies to assess the overall utility of the PIVO device are warranted. PMID:28419012

  3. Biomarkers for Monitoring Pre-Analytical Quality Variation of mRNA in Blood Samples

    PubMed Central

    Zhang, Hui; Korenková, Vlasta; Sjöback, Robert; Švec, David; Björkman, Jens; Kruhøffer, Mogens; Verderio, Paolo; Pizzamiglio, Sara; Ciniselli, Chiara Maura; Wyrich, Ralf; Oelmueller, Uwe; Kubista, Mikael; Lindahl, Torbjørn; Lönneborg, Anders; Rian, Edith

    2014-01-01

    There is an increasing need for proper quality control tools in the pre-analytical phase of the molecular diagnostic workflow. The aim of the present study was to identify biomarkers for monitoring pre-analytical mRNA quality variations in two different types of blood collection tubes, K2EDTA (EDTA) tubes and PAXgene Blood RNA Tubes (PAXgene tubes). These tubes are extensively used both in the diagnostic setting as well as for research biobank samples. Blood specimens collected in the two different blood collection tubes were stored for varying times at different temperatures, and microarray analysis was performed on resultant extracted RNA. A large set of potential mRNA quality biomarkers for monitoring post-phlebotomy gene expression changes and mRNA degradation in blood was identified. qPCR assays for the potential biomarkers and a set of relevant reference genes were generated and used to pre-validate a sub-set of the selected biomarkers. The assay precision of the potential qPCR based biomarkers was determined, and a final validation of the selected quality biomarkers using the developed qPCR assays and blood samples from 60 healthy additional subjects was performed. In total, four mRNA quality biomarkers (USP32, LMNA, FOSB, TNRFSF10C) were successfully validated. We suggest here the use of these blood mRNA quality biomarkers for validating an experimental pre-analytical workflow. These biomarkers were further evaluated in the 2nd ring trial of the SPIDIA-RNA Program which demonstrated that these biomarkers can be used as quality control tools for mRNA analyses from blood samples. PMID:25369468

  4. Method and apparatus for automated processing and aliquoting of whole blood samples for analysis in a centrifugal fast analyzer

    DOEpatents

    Burtis, Carl A.; Johnson, Wayne F.; Walker, William A.

    1988-01-01

    A rotor and disc assembly for use in a centrifugal fast analyzer. The assembly is designed to process multiple samples of whole blood followed by aliquoting of the resultant serum into precisely measured samples for subsequent chemical analysis. The assembly requires minimal operator involvement with no mechanical pipetting. The system comprises (1) a whole blood sample disc, (2) a serum sample disc, (3) a sample preparation rotor, and (4) an analytical rotor. The blood sample disc and serum sample disc are designed with a plurality of precision bore capillary tubes arranged in a spoked array. Samples of blood are loaded into the blood sample disc in capillary tubes filled by capillary action and centrifugally discharged into cavities of the sample preparation rotor where separation of serum and solids is accomplished. The serum is loaded into the capillaries of the serum sample disc by capillary action and subsequently centrifugally expelled into cuvettes of the analytical rotor for analysis by conventional methods.

  5. Method and apparatus for automated processing and aliquoting of whole blood samples for analysis in a centrifugal fast analyzer

    DOEpatents

    Burtis, C.A.; Johnson, W.F.; Walker, W.A.

    1985-08-05

    A rotor and disc assembly for use in a centrifugal fast analyzer. The assembly is designed to process multiple samples of whole blood followed by aliquoting of the resultant serum into precisely measured samples for subsequent chemical analysis. The assembly requires minimal operator involvement with no mechanical pipetting. The system comprises: (1) a whole blood sample disc; (2) a serum sample disc; (3) a sample preparation rotor; and (4) an analytical rotor. The blood sample disc and serum sample disc are designed with a plurality of precision bore capillary tubes arranged in a spoked array. Samples of blood are loaded into the blood sample disc by capillary action and centrifugally discharged into cavities of the sample preparation rotor where separation of serum and solids is accomplished. The serum is loaded into the capillaries of the serum sample disc by capillary action and subsequently centrifugally expelled into cuvettes of the analyticaly rotor for conventional methods. 5 figs.

  6. A cell transportation solution that preserves live circulating tumor cells in patient blood samples.

    PubMed

    Stefansson, Steingrimur; Adams, Daniel L; Ershler, William B; Le, Huyen; Ho, David H

    2016-05-06

    Circulating tumor cells (CTCs) are typically collected into CellSave fixative tubes, which kills the cells, but preserves their morphology. Currently, the clinical utility of CTCs is mostly limited to their enumeration. More detailed investigation of CTC biology can be performed on live cells, but obtaining live CTCs is technically challenging, requiring blood collection into biocompatible solutions and rapid isolation which limits transportation options. To overcome the instability of CTCs, we formulated a sugar based cell transportation solution (SBTS) that stabilizes cell viability at ambient temperature. In this study we examined the long term viability of human cancer cell lines, primary cells and CTCs in human blood samples in the SBTS for transportation purposes. Four cell lines, 5 primary human cells and purified human PBMCs were tested to determine the viability of cells stored in the transportation solution at ambient temperature for up to 7 days. We then demonstrated viability of MCF-7 cells spiked into normal blood with SBTS and stored for up to 7 days. A pilot study was then run on blood samples from 3 patients with metastatic malignancies stored with or without SBTS for 6 days. CTCs were then purified by Ficoll separation/microfilter isolation and identified using CTC markers. Cell viability was assessed using trypan blue or CellTracker™ live cell stain. Our results suggest that primary/immortalized cell lines stored in SBTS remain ~90% viable for > 72 h. Further, MCF-7 cells spiked into whole blood remain viable when stored with SBTS for up to 7 days. Finally, live CTCs were isolated from cancer patient blood samples kept in SBTS at ambient temperature for 6 days. No CTCs were isolated from blood samples stored without SBTS. In this proof of principle pilot study we show that viability of cell lines is preserved for days using SBTS. Further, this solution can be used to store patient derived blood samples for eventual isolation of viable CTCs after

  7. Direct and long-term detection of gene doping in conventional blood samples.

    PubMed

    Beiter, T; Zimmermann, M; Fragasso, A; Hudemann, J; Niess, A M; Bitzer, M; Lauer, U M; Simon, P

    2011-03-01

    The misuse of somatic gene therapy for the purpose of enhancing athletic performance is perceived as a coming threat to the world of sports and categorized as 'gene doping'. This article describes a direct detection approach for gene doping that gives a clear yes-or-no answer based on the presence or absence of transgenic DNA in peripheral blood samples. By exploiting a priming strategy to specifically amplify intronless DNA sequences, we developed PCR protocols allowing the detection of very small amounts of transgenic DNA in genomic DNA samples to screen for six prime candidate genes. Our detection strategy was verified in a mouse model, giving positive signals from minute amounts (20 μl) of blood samples for up to 56 days following intramuscular adeno-associated virus-mediated gene transfer, one of the most likely candidate vector systems to be misused for gene doping. To make our detection strategy amenable for routine testing, we implemented a robust sample preparation and processing protocol that allows cost-efficient analysis of small human blood volumes (200 μl) with high specificity and reproducibility. The practicability and reliability of our detection strategy was validated by a screening approach including 327 blood samples taken from professional and recreational athletes under field conditions.

  8. Fetal blood sampling in baboons (Papio spp.): important procedural aspects and literature review.

    PubMed

    Joy, S D; O'Shaughnessy, R; Schlabritz-Loutsevitch, N; Leland, M M; Frost, P; Fan-Havard, P

    2009-06-01

    The baboons (Papio cynocephalus) have similarities with human placentation and fetal development. Fetal blood sampling allows investigators to assess fetal condition at a specific point in gestation as well as transplacental transfer of medications. Unfortunately, assessing fetal status during gestation has been difficult and fetal instrumentation associated with high rate of pregnancy loss. Our objectives are to describe the technique of ultrasound guided cordocentesis (UGC) in baboons, report post-procedural outcomes, and review existing publications. This is a procedural paper describing the technique of UGC in baboons. After confirming pregnancy and gestational age via ultrasound, animals participating in approved research protocols that required fetal assessment underwent UGC. We successfully performed UGC in four animals (five samples) using this technique. Animals were sampled in the second and third trimesters with fetal blood sampling achieved by sampling a free cord loop, placental cord insertion site or the intrahepatic umbilical vein. All procedures were without complication and these animals delivered at term. Ultrasound guided fetal umbilical cord venipuncture is a useful and safe technique to sample the fetal circulation with minimal risk to the fetus or mother. We believe this technique could be used for repeated fetal venous blood sampling in the baboons.

  9. 1-Hydroxypyrene Levels in Blood Samples of Rats After Exposure to Generator Fumes

    PubMed Central

    Ifegwu, Clinton; Igwo-Ezikpe, Miriam N.; Anyakora, Chimezie; Osuntoki, Akinniyi; Oseni, Kafayat A.; Alao, Eragbae O.

    2013-01-01

    Polynuclear Aromatic Hydrocarbons (PAHs) are a major component of fuel generator fumes. Carcinogenicity of these compounds has long been established. In this study, 37 Swiss albino rats were exposed to generator fumes at varied distances for 8 hours per day for a period of 42 days and the level of 1-hydroxypyrene in their blood was evaluated. This study also tried to correlate the level of blood 1-hyroxypyrene with the distance from the source of pollution. Plasma was collected by centrifuging the whole blood sample followed by complete hydrolysis of the conjugated 1-hydroxypyrene glucuronide to yield the analyte of interest, 1-hydroxypyrene, which was achieved using beta glucuronidase. High performance liquid chromatography (HPLC) with UV detector was used to determine the 1-hydroxypyrene concentrations in the blood samples. The mobile phase was water:methanol (12:88 v/v) isocratic run at the flow rate of 1.2 mL/min with CI8 stationary phase at 250 nm. After 42 days of exposure, blood concentration level of 1-hydroxypyrene ranged from 34 μg/mL to 26.29 μg/mL depending on the distance from source of exposure. The control group had no 1-hydroxypyrene in their blood. After the period of exposure, percentage of death correlated with the distance from the source of exposure. Percentage of death ranged from 56% to zero depending on the proximity to source of pollution. PMID:24179393

  10. 1-hydroxypyrene levels in blood samples of rats after exposure to generator fumes.

    PubMed

    Ifegwu, Clinton; Igwo-Ezikpe, Miriam N; Anyakora, Chimezie; Osuntoki, Akinniyi; Oseni, Kafayat A; Alao, Eragbae O

    2013-01-01

    Polynuclear Aromatic Hydrocarbons (PAHs) are a major component of fuel generator fumes. Carcinogenicity of these compounds has long been established. In this study, 37 Swiss albino rats were exposed to generator fumes at varied distances for 8 hours per day for a period of 42 days and the level of 1-hydroxypyrene in their blood was evaluated. This study also tried to correlate the level of blood 1-hyroxypyrene with the distance from the source of pollution. Plasma was collected by centrifuging the whole blood sample followed by complete hydrolysis of the conjugated 1-hydroxypyrene glucuronide to yield the analyte of interest, 1-hydroxypyrene, which was achieved using beta glucuronidase. High performance liquid chromatography (HPLC) with UV detector was used to determine the 1-hydroxypyrene concentrations in the blood samples. The mobile phase was water:methanol (12:88 v/v) isocratic run at the flow rate of 1.2 mL/min with CI8 stationary phase at 250 nm. After 42 days of exposure, blood concentration level of 1-hydroxypyrene ranged from 34 μg/mL to 26.29 μg/mL depending on the distance from source of exposure. The control group had no 1-hydroxypyrene in their blood. After the period of exposure, percentage of death correlated with the distance from the source of exposure. Percentage of death ranged from 56% to zero depending on the proximity to source of pollution.

  11. A novel first principles approach for the estimation of the sieve factor of blood samples.

    PubMed

    Northam, L; Baranoski, G V G

    2010-03-29

    Light may traverse a turbid material, such as blood, without encountering any of its pigment containing structures, a phenomenon known as sieve effect. This phenomenon may result in a decrease in the amount of light absorbed by the material. Accordingly, the corresponding sieve factor needs to be accounted for in optical investigations aimed at the derivation of blood biophysical properties from light transmittance measurements. The existing procedures used for its estimation either lack the flexibility required for practical applications or are based on general formulas that incorporate other light and matter interaction phenomena such as detour (scattering) effects. In this paper, a ray optics framework is proposed to estimate the sieve factor for blood samples. It employs a first principles approach to account for the distribution, orientation and shape of the cells that contain hemoglobin, the essential (oxygen-carrying) pigment found in human blood. Within this framework, ray-casting techniques are used to determine the probability that light can traverse a blood sample without encountering any of these cells. The predictive capabilities of the proposed framework are demonstrated through a series of in silico experiments. Its effectiveness is further illustrated by visualizations depicting the different blood parameterizations considered in the simulations.

  12. Detection of the BLV provirus from nasal secretion and saliva samples using BLV-CoCoMo-qPCR-2: Comparison with blood samples from the same cattle.

    PubMed

    Yuan, Yuan; Kitamura-Muramatsu, Yuri; Saito, Susumu; Ishizaki, Hiroshi; Nakano, Miwa; Haga, Satoshi; Matoba, Kazuhiro; Ohno, Ayumu; Murakami, Hironobu; Takeshima, Shin-Nosuke; Aida, Yoko

    2015-12-02

    Bovine leukemia virus (BLV) induces enzootic bovine leukosis, which is the most common neoplastic disease in cattle. Sero-epidemiological studies show that BLV infection occurs worldwide. Direct contact between infected and uninfected cattle is thought to be one of the risk factors for BLV transmission. Contact transmission occurs via a mixture of natural sources, blood, and exudates. To confirm that BLV provirus is detectable in these samples, matched blood, nasal secretion, and saliva samples were collected from 50 cattle, and genomic DNA was extracted. BLV-CoCoMo-qPCR-2, an assay developed for the highly sensitive detection of BLV, was then used to measure the proviral load in blood (n=50), nasal secretions (n=48), and saliva (n=47) samples. The results showed that 35 blood samples, 14 nasal secretion samples, and 6 saliva samples were positive for the BLV provirus. Matched blood samples from cattle that were positive for the BLV provirus (either in nasal secretion or saliva samples) were also positive in their blood. The proviral load in the positive blood samples was >14,000 (copies/1×10(5) cells). Thus, even though the proviral load in the nasal secretion and saliva samples was much lower (<380 copies/1×10(5) cells) than that in the peripheral blood, prolonged direct contact between infected and healthy cattle may be considered as a risk factor for BLV transmission.

  13. Effective prevention of pseudothrombocytopenia in feline blood samples with the prostaglandin I2 analogue Iloprost.

    PubMed

    Riond, Barbara; Waßmuth, Andrea Katharina; Hartnack, Sonja; Hofmann-Lehmann, Regina; Lutz, Hans

    2015-08-06

    In vitro platelet aggregation in feline blood samples is a well-known phenomenon in veterinary clinical laboratories resulting in high numbers of pseudothrombocytopenia. Several attempts have been made to prevent or dissolve platelet aggregates in feline blood samples and to increase the reliability of feline platelet counts. Prostaglandin I2 (PGI2) is the most powerful endogenous inhibitor of platelet aggregation but unstable. Iloprost is a stable PGI2 analogue. The aims of the present study were (1) to evaluate the anti-aggregatory effect of Iloprost on feline platelet counts and to determine a useful concentration to inhibit platelet aggregation in EDTA samples from clinically healthy cats, (2) to investigate the effect of Iloprost on hematological blood parameters, and (3) to determine stability of Iloprost in K3-EDTA tubes for up to 16 weeks. From 20 clinically healthy cats blood was drawn from the jugular vein and immediately distributed in a 1.3 ml K3-EDTA tube, and two 1.3 ml K3-EDTA tubes containing 20 ng and 200 ng Iloprost, respectively. A complete blood cell count was performed on the Sysmex XT-2000iV and the Mythic 18 on eight consecutive time points after collection. Blood smears were evaluated for the presence of PLT aggregates. In the absence of Iloprost, pseudothrombocytopenia was observed in 50% of the investigated samples that led to significantly decreased optical PLT counts by a mean of 105 x10(3)/μl, which could be prevented by the addition of 1 μL (20 ng) Iloprost leading to an increase in PLT counts by a mean of 108 x10(3)/μl. This is the first study showing an anti-aggregatory effect of the PGI2-analogue Iloprost in feline EDTA blood. In all clinically healthy cats investigated, pseudothrombocytopenia was prevented by adding Iloprost to EDTA tubes prior to blood collection. Furthermore, Iloprost was very useful in preventing falsely increased WBC counts in samples with platelet aggregates analyzed on impedance-based hematological

  14. Blood, sweat, and tears: embedding biological samples in social science research on children.

    PubMed

    Kall, Denise

    2008-01-01

    In the first decade of the 21st Century, calls for interdisciplinary research are commonplace. Yet, relatively few papers discuss how to complete such research successfully. In this paper, I describe the details of data collection focused on five, six and seven-year old children. The project examined the effect of environmental contaminants on children's educational outcomes. It included a primary caregiver interview, a skill test with the child, and a venous blood draw from the child to test for lead, mercury, cadmium, arsenic, nicotine, and cotinine. This paper describes key issues and the solutions I adopted. Challenges discussed here include navigating the Institutional Review Board Process, analyzing the blood, obtaining the supplies needed to draw blood, banking blood for future research, hiring a phlebotomist, and recruiting subjects. While not all details will apply directly to other research projects, this paper provides some perspective on the current realities facing social scientists who decide to collect biological samples.

  15. In vitro study of thimerosal reactions in human whole blood and plasma surrogate samples.

    PubMed

    Trümpler, Stefan; Meermann, Björn; Nowak, Sascha; Buscher, Wolfgang; Karst, Uwe; Sperling, Michael

    2014-04-01

    Because of its bactericidal and fungicidal properties, thimerosal is used as a preservative in drugs and vaccines and is thus deliberately injected into the human body. In aqueous environment, it decomposes into thiosalicylic acid and the ethylmercury cation. This organomercury fragment is a potent neurotoxin and is suspected to have similar toxicity and bioavailability like the methylmercury cation. In this work, human whole blood and physiological simulation solutions were incubated with thimerosal to investigate its behaviour and binding partners in the blood stream. Inductively coupled plasma with optical emission spectrometry (ICP-OES) was used for total mercury determination in different blood fractions, while liquid chromatography (LC) coupled to electrospray ionisation time-of-flight (ESI-TOF) and inductively coupled plasma-mass spectrometry (ICP-MS) provided information on the individual mercury species in plasma surrogate samples. Analogous behaviour of methylmercury and ethylmercury species in human blood was shown and an ethylmercury-glutathione adduct was identified.

  16. The effect of delayed transportation of blood samples on serum bilirubin values in neonates.

    PubMed

    Saththasivam, Poovendran; Voralu, Kirtanaa; Ramli, Noraida; Mustapha, Mohd Rafi; Omar, Julia; Van Rostenberghe, Hans

    2010-07-01

    Delays in transporting blood samples may cause inaccurate results. Samples may be exposed to light or heat during delays, resulting in the degradation of analytes, for example, bilirubin. This study was done to determine the effect of delays in the transportation of blood samples on serum bilirubin test results. Samples taken from neonates admitted to a tertiary hospital with jaundice were included in the study. The samples were collected through venipuncture in 3 labelled containers. The first container was sent immediately to the laboratory, while the second and third containers were sent after being kept in the ward for 1 and 3 hours, respectively. Bilirubin values were measured colourimetrically at a wavelength of 578 nm using a Roche Hitachi 912 Chemistry Analyser upon arrival in the laboratory. A total of 36 serum samples were studied. The mean of the indirect bilirubin measurements for 0-, 1-, and 3-hour samples were 174 (SD 68.65), 186.97 (SD 60.47), and 184.56 (SD 66.93), respectively. There was a significant difference in the mean indirect bilirubin measurement of 1-hour samples (P = 0.047, 95% CI -24.66 to -1.18) and 3-hour samples (P = 0.045, 95% CI -19.77 to -0.23) compared with 0-hour samples. There were no significant differences observed in either the mean total bilirubin or the mean direct bilirubin measurements of different time intervals. This study confirms that delays in the transportation of blood samples influence the bilirubin test results.

  17. Small and cheap: accurate differential blood count with minimal sample volume by laser scanning cytometry (LSC)

    NASA Astrophysics Data System (ADS)

    Mittag, Anja; Lenz, Dominik; Smith, Paul J.; Pach, Susanne; Tarnok, Attila

    2005-04-01

    Aim: In patients, e.g. with congenital heart diseases, a differential blood count is needed for diagnosis. To this end by standard automatic analyzers 500 μl of blood is required from the patients. In case of newborns and infants this is a substantial volume, especially after operations associated with blood loss. Therefore, aim of this study was to develop a method to determine a differential blood picture with a substantially reduced specimen volume. Methods: To generate a differential blood picture 10 μl EDTA blood were mixed with 10 μl of a DRAQ5 solution (500μM, Biostatus) and 10 μl of an antibody mixture (CD45-FITC, CD14-PE, diluted with PBS). 20 μl of this cell suspension was filled into a Neubauer counting chamber. Due to the defined volume of the chamber it is possible to determine the cell count per volume. The trigger for leukocyte counting was set on DRAQ5 signal in order to be able to distinguish nucleated white blood cells from erythrocytes. Different leukocyte subsets could be distinguished due to the used fluorescence labeled antibodies. For erythrocyte counting cell suspension was diluted another 150 times. 20 μl of this dilution was analyzed in a microchamber by LSC with trigger set on forward scatter signal. Results: This method allows a substantial decrease of blood sample volume for generation of a differential blood picture (10 μl instead of 500μl). There was a high correlation between our method and the results of routine laboratory (r2=0.96, p<0.0001 n=40). For all parameters intra-assay variance was less than 7 %. Conclusions: In patients with low blood volume such as neonates and in critically ill infants every effort has to be taken to reduce the blood volume needed for diagnostics. With this method only 2% of standard sample volume is needed to generate a differential blood picture. Costs are below that of routine laboratory. We suggest this method to be established in paediatric cardiology for routine diagnostics and for

  18. Risk factors for Salmonella infection in fattening pigs - an evaluation of blood and meat juice samples.

    PubMed

    Hotes, S; Kemper, N; Traulsen, I; Rave, G; Krieter, J

    2010-11-01

    The main objective of this study was to analyse potential herd-level factors associated with the detection of Salmonella antibodies in fattening pigs. Two independent datasets, consisting of blood and meat juice samples respectively, were used. Additional information about husbandry, management and hygiene conditions was collected by questionnaire for both datasets. The serological analysis showed that 13.8% of the blood samples and 15.7% of the meat juice samples had to be classified as Salmonella-positive. Logistic-regression models were used to assess statistically significant risk factors associated with a positive sample result. The results of the statistical blood sample analysis showed that the application of antibiotics increased the odds ratio (OR) by a factor of 5.21 (P < 0.001) compared to untreated pigs. A fully slatted floor decreased the prevalence of Salmonella as well as the use of protective clothing or the cleaning of the feed tube (ORs 0.35-0.54, P < 0.001). It was shown that a distance of less than 2 km to other swine herds increased the chance of a positive Salmonella result (OR = 3.76, P < 0.001). The statistical analysis of the meat juice samples revealed the importance of feed aspects. The chance of obtaining a positive meat juice sample increased by a factor of 3.52 (P < 0.001) by using granulated feed instead of flour. It also became clear that liquid feeding should be preferred to dry feeding (OR = 0.33, P < 0.001). A comparison of the blood sample analysis to the meat juice model revealed that the latter was less powerful because data structure was less detailed. The expansion of data acquisition might solve these problems and improve the suitability of QS monitoring data for risk factor analyses. © 2010 Blackwell Verlag GmbH.

  19. Comparison of manual and automated nucleic acid extraction from whole-blood samples.

    PubMed

    Riemann, Kathrin; Adamzik, Michael; Frauenrath, Stefan; Egensperger, Rupert; Schmid, Kurt W; Brockmeyer, Norbert H; Siffert, Winfried

    2007-01-01

    Nucleic acid extraction and purification from whole blood is a routine application in many laboratories. Automation of this procedure promises standardized sample treatment, a low error rate, and avoidance of contamination. The performance of the BioRobot M48 (Qiagen) and the manual QIAmp DNA Blood Mini Kit (Qiagen) was compared for the extraction of DNA from whole blood. The concentration and purity of the extracted DNAs were determined by spectrophotometry. Analytical sensitivity was assessed by common PCR and genotyping techniques. The quantity and quality of the generated DNAs were slightly higher using the manual extraction method. The results of downstream applications were comparable to each other. Amplification of high-molecular-weight PCR fragments, genotyping by restriction digest, and pyrosequencing were successful for all samples. No cross-contamination could be detected. While automated DNA extraction requires significantly less hands-on time, it is slightly more expensive than the manual extraction method.

  20. Blood culture sampling rates at a German pediatric university hospital and incidence of invasive pneumococcal disease.

    PubMed

    Rüggeberg, J U; Ketteler, K; MacKenzie, C R; Von Kries, R; Reinert, R R; Schroten, H

    2004-04-01

    Recent pediatric surveillance studies suggest the incidence of pneumococcal bacteremia, but not meningitis, is lower in Germany than in most developed countries. Suboptimal case assessment in routine clinical practice has been suspected of contributing to this apparent discrepancy. We aimed to assess the blood culture sampling rate at a German pediatric university hospital and the disease burden associated with pneumococcal bacteremia in children under 5 years of age. The study design was retrospective, based on data-linkage and chart review. Blood cultures were frequently obtained in sepsis (96%; CI 78-99%) and meningitis (95%; CI 77-99%), but less commonly in pneumonia (49%; CI 43-54%) and fever without focus (48%; CI 38-59%). Pneumococci were the most common source of clinically significant bacteremia in previously healthy children. These blood culture sampling rates may be insufficient for the sensitive detection of pneumococcal bacteremia. Epidemiological surveillance based on poorly standardized diagnostic practices is prone to under-assessment.

  1. Application of automated serial blood sampling and dried blood spot technique with liquid chromatography-tandem mass spectrometry for pharmacokinetic studies in mice.

    PubMed

    Wong, Philip; Pham, Roger; Whitely, Carl; Soto, Marcus; Salyers, Kevin; James, Christopher; Bruenner, Bernd A

    2011-11-01

    The goal of this work was to obtain full pharmacokinetic profiles from individual mice with the use of an automated blood sampling system and dried blood spot (DBS) technique. AMG 517, a potent and selective vanilloid receptor (VR1) antagonist, was dosed to mice (n=3) intravenously and blood samples were collected using the automated blood sampling system with the "no blood waste" method. The collected blood samples were a mixture of 25 μL blood and 50 μL of heparinized saline solution. Two 15 μL aliquots were manually spotted onto a DBS card and dried at room temperature for at least 2h before being stored in zip bags with desiccant. The remaining samples (45 μL) were stored at -70°C until analysis. Both the DBS and the whole blood samples (diluted with saline (1:2, v/v)) were extracted and analyzed by liquid chromatography-tandem mass spectrometry. The overall extraction recovery of the analyte from the dried blood spots was determined to be about 90%. The pharmacokinetic parameters calculated using the whole blood or the DBS concentration data were comparable, and were obtained from only 3 mice, whereas conventional sampling and analysis would have required up to 27 mice to achieve the same result. The analyte was shown to be stable in the diluted whole blood (blood:saline 1:2) at room temperature for at least 4h and in the DBS for at least 34 days when stored at room temperature. These results indicated that the automated blood sampling system and DBS collection are promising techniques to obtain full pharmacokinetic profiles from individual mice and reduce the use of animals. Copyright © 2011. Published by Elsevier B.V.

  2. Adapting dried blood spot sampling for an anti-therapeutic antibody immunogenicity assay.

    PubMed

    Xiang, Yuhong; Welch, Mackenzie; Amaravadi, Lakshmi; Stebbins, Christopher

    2013-07-31

    Dried blood spot sampling is a microvolume sampling technique with many potential advantages. It allows for easier handling and less expensive shipment and storage of biological samples. Additionally, it can provide ethical benefits in the pre-clinical setting through a reduction in animal usage by allowing intensive serial sample collection from the same animals. In the clinical setting, ease of sample collection, greater flexibility of sample storage, and shipping are distinct advantages. These advantages can enhance preclinical and clinical data quality, where immunogenicity monitoring plays an important role in the interpretation of pharmacokinetic data. To date, a method for usage of dried blood spot sampling with an immunogenicity assay has not been published. Herein we demonstrate that the measurement of anti-drug antibodies (ADA) using DBS was comparable to traditional methods in terms of reproducibility, assay sensitivity and drug tolerance. The data demonstrate that DBS is a viable sample collection method, and in some cases may be preferred, over classic serum or plasma sampling for antidrug antibody assays. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Transcutaneous bilirubinometry reduces the need for blood sampling in neonates with visible jaundice.

    PubMed

    Mishra, S; Chawla, D; Agarwal, R; Deorari, A K; Paul, V K; Bhutani, V K

    2009-12-01

    We determined usefulness of transcutaneous bilirubinometry to decrease the need for blood sampling to assay serum total bilirubin (STB) in the management of jaundiced healthy Indian neonates. Newborns, > or =35 weeks' gestation, with clinical evidence of jaundice were enrolled in an institutional approved randomized clinical trial. The severity of hyperbilirubinaemia was determined by two non-invasive methods: i) protocol-based visual assessment of bilirubin (VaB) and ii) transcutaneous bilirubin (TcB) determination (BiliCheck). By a random allocation, either method was used to decide the need for blood sampling, which was defined to be present if assessed STB by allocated method exceeded 80% of hour-specific threshold values for phototherapy (2004 AAP Guidelines). A total of 617 neonates were randomized to either TcB (n = 314) or VaB (n = 303) groups with comparable gestation, birth weight and postnatal age. Need for blood sampling to assay STB was 34% lower (95% CI: 10% to 51%) in the TcB group compared with VaB group (17.5% vs 26.4% assessments; risk difference: -8.9%, 95% CI: -2.4% to -15.4%; p = 0.008). Routine use of transcutaneous bilirubinometry compared with systematic visual assessment of bilirubin significantly reduced the need for blood sampling to assay STB in jaundiced term and late-preterm neonates. (ClinicalTrials.gov number, NCT00653874).

  4. Alkylphenol Polyethoxylate Derivatives in Groundwater and Blood Samples Collected from Pig Herds in Taiwan

    PubMed Central

    CHIU, Tai-Shun; HSIEH, Chi-Ying; MIAW, Chang-Ling; LIN, Chao-Nan; CHANG, Tsung-Chou; YEN, Chia-Hung; CHIOU, Ming-Tang

    2014-01-01

    ABSTRACT Alkylphenol polyethoxylate (APEO) derivatives, such as nonylphenol monoethoxylate (NP1EO), nonylphenol diethoxylate (NP2EO), nonylphenol (NP) and octylphenol (OP), have been detected in the surface water, sediment, food and groundwater of numerous countries. Because groundwater is the main source of water for pig herds, the aim of this study was to measure the concentrations of APEO derivatives in groundwater and blood samples that were collected from pig herds raised near the Wuluo River in Southern Taiwan. The mean concentrations of NP, OP, NP1EO and NP2EO in the groundwater supply for 10 pig herds were 0.04 µg/l, 0.26 ± 0.23 µg/l, 0.74 ± 0.69 µg/l and 0.17 ± 0.22 µg/l, respectively. NP was detected in all blood samples collected from 5 of the 10 pig herds. The highest concentrations detected in the blood samples collected from six-week-old piglets and sows were 12.00 µg/l and 56.94 µg/l, respectively. Blood samples from 4 of the 5 herds showed OP contamination. The highest OP concentrations detected in 6-week-old piglets and sows were 275.58 µg/l and 566.32 µg/l, respectively. These results indicate that APEO derivatives accumulated in the groundwater supply and the bloodstreams of the pigs. PMID:24694943

  5. Alkylphenol polyethoxylate derivatives in groundwater and blood samples collected from pig herds in Taiwan.

    PubMed

    Chiu, Tai-Shun; Hsieh, Chi-Ying; Miaw, Chang-Ling; Lin, Chao-Nan; Chang, Tsung-Chou; Yen, Chia-Hung; Chiou, Ming-Tang

    2014-07-01

    Alkylphenol polyethoxylate (APEO) derivatives, such as nonylphenol monoethoxylate (NP1EO), nonylphenol diethoxylate (NP2EO), nonylphenol (NP) and octylphenol (OP), have been detected in the surface water, sediment, food and groundwater of numerous countries. Because groundwater is the main source of water for pig herds, the aim of this study was to measure the concentrations of APEO derivatives in groundwater and blood samples that were collected from pig herds raised near the Wuluo River in Southern Taiwan. The mean concentrations of NP, OP, NP1EO and NP2EO in the groundwater supply for 10 pig herds were 0.04 µg/l, 0.26 ± 0.23 µg/l, 0.74 ± 0.69 µg/l and 0.17 ± 0.22 µg/l, respectively. NP was detected in all blood samples collected from 5 of the 10 pig herds. The highest concentrations detected in the blood samples collected from six-week-old piglets and sows were 12.00 µg/l and 56.94 µg/l, respectively. Blood samples from 4 of the 5 herds showed OP contamination. The highest OP concentrations detected in 6-week-old piglets and sows were 275.58 µg/l and 566.32 µg/l, respectively. These results indicate that APEO derivatives accumulated in the groundwater supply and the bloodstreams of the pigs.

  6. Blood sampling in juvenile buff-breasted sandpipers: Movement, weight change and survival

    USGS Publications Warehouse

    Lanctot, Richard B.

    1994-01-01

    The effect of blood sampling on juvenile Buff-breasted Sandpipers (Tryngites subruficollis) was evaluated by comparing movements, mass, and survival of 10 broods (37 chicks) that were bled and eight broods (31 chicks) that were not bled. Blood was sampled from the jugular vein of chicks when they weighed 9.1 ± 0.9 g (x̄ ± SD) on or within 1 d of hatch. Chicks showed few short-term negative effects from blood sampling. Individual chicks suffered little physical injury, and five of eight chicks where injury occurred (i.e., hematomas formed) survived to fledging. Furthermore, bled broods gained mass at a comparable rate during the first 5 d post-hatch, and were resighted at similar frequencies as broods that were not bled. Bled broods moved slightly longer distances than control broods 1 d after hatch, however. This increased activity may have been stress-induced, but was only temporary; bled and control broods made similar long-term movements, and the probability of resighting was similar at fledging. With the proper precautions, it appears that Buff-breasted Sandpiper young can be safely sampled for blood at an early age without causing undue harm.

  7. Novel genotype of Ehrlichia canis detected in samples of human blood bank donors in Costa Rica.

    PubMed

    Bouza-Mora, Laura; Dolz, Gaby; Solórzano-Morales, Antony; Romero-Zuñiga, Juan José; Salazar-Sánchez, Lizbeth; Labruna, Marcelo B; Aguiar, Daniel M

    2017-01-01

    This study focuses on the detection and identification of DNA and antibodies to Ehrlichia spp. in samples of blood bank donors in Costa Rica using molecular and serological techniques. Presence of Ehrlichia canis was determined in 10 (3.6%) out of 280 blood samples using polymerase chain reaction (PCR) targeting the ehrlichial dsb conserved gene. Analysis of the ehrlichial trp36 polymorphic gene in these 10 samples revealed substantial polymorphism among the E. canis genotypes, including divergent tandem repeat sequences. Nucleotide sequences of dsb and trp36 amplicons revealed a novel genotype of E. canis in blood bank donors from Costa Rica. Indirect immunofluorescence assay (IFA) detected antibodies in 35 (35%) of 100 serum samples evaluated. Thirty samples showed low endpoint titers (64-256) to E. canis, whereas five sera yielded high endpoint titers (1024-8192); these five samples were also E. canis-PCR positive. These findings represent the first report of the presence of E. canis in humans in Central America. Copyright © 2016 Elsevier GmbH. All rights reserved.

  8. Prior exercise alters the difference between arterialised and venous glycaemia: implications for blood sampling procedures.

    PubMed

    Edinburgh, Robert M; Hengist, Aaron; Smith, Harry A; Betts, James A; Thompson, Dylan; Walhin, Jean-Philippe; Gonzalez, Javier T

    2017-05-01

    Oral glucose tolerance and insulin sensitivity are common measures, but are determined using various blood sampling methods, employed under many different experimental conditions. This study established whether measures of oral glucose tolerance and oral glucose-derived insulin sensitivity (insulin sensitivity indices; ISI) differ when calculated from venous v. arterialised blood. Critically, we also established whether any differences between sampling methods are consistent across distinct metabolic conditions (after rest v. after exercise). A total of ten healthy men completed two trials in a randomised order, each consisting of a 120-min oral glucose tolerance test (OGTT), either at rest or post-exercise. Blood was sampled simultaneously from a heated hand (arterialised) and an antecubital vein of the contralateral arm (venous). Under both conditions, glucose time-averaged AUC was greater from arterialised compared with venous plasma but importantly, this difference was larger after rest relative to after exercise (0·99 (sd 0·46) v. 0·56 (sd 0·24) mmol/l, respectively; P<0·01). OGTT-derived ISIMatsuda and ISICederholm were lower when calculated from arterialised relative to venous plasma and the arterialised-venous difference was greater after rest v. after exercise (ISIMatsuda: 1·97 (sd 0·81) v. 1·35 (sd 0·57) arbitrary units (au), respectively; ISICederholm : 14·76 (sd 7·83) v. 8·70 (sd 3·95) au, respectively; both P<0·01). Venous blood provides lower postprandial glucose concentrations and higher estimates of insulin sensitivity, compared with arterialised blood. Most importantly, these differences between blood sampling methods are not consistent after rest v. post-exercise, preventing standardised venous-to-arterialised corrections from being readily applied.

  9. Comparison of three methods for recovery of Brucella canis DNA from canine blood samples.

    PubMed

    Batinga, Maria Cryskely A; Dos Santos, Jaíne C; Lima, Julia T R; Bigotto, Maria Fernanda D; Muner, Kerstin; Faita, Thalita; Soares, Rodrigo M; da Silva, David A V; Oliveira, Trícia M F S; Ferreira, Helena L; Diniz, Jaqueline A; Keid, Lara B

    2017-08-29

    Brucella canis, a gram-negative, facultative intracellular and zoonotic bacterium causes canine brucellosis. Direct methods are the most appropriate for the detection of canine brucellosis and bacterial isolation from blood samples has been employed as gold-standard method. However, due to the delay in obtaining results and the biological risk of the bacterial culturing, the polymerase chain reaction (PCR) has been successfully used as an alternative method for the diagnosis of the infection. Sample preparation is a key step for successful PCR and protocols that provide high DNA yield and purity are recommended to ensure high diagnostic sensitivity. The objective of this study was to evaluate the performance of PCR for the diagnosis of B. canis infection in 36 dogs by testing DNA of whole blood obtained through different extraction and purification protocols. Methods 1 and 2 were based on a commercial kit, using protocols recommended for DNA purification of whole blood and tissue samples, respectively. Method 3 was an in-house method based on enzymatic lysis and purification using organic solvents. The results of the PCR on samples obtained through three different DNA extraction protocols were compared to the blood culture. Of the 36 dogs, 13 (36.1%) were positive by blood culturing, while nine (25.0%), 14 (38.8%), and 15 (41.6%) were positive by PCR after DNA extraction using methods 1, 2 and 3, respectively. PCR performed on DNA purified by Method 2 was as efficient as blood culturing and PCR performed on DNA purified with in-house method, but had the advantage of being less laborious and, therefore, a suitable alternative for the direct B. canis detection in dogs. Copyright © 2017. Published by Elsevier B.V.

  10. Application of the Triage panel for drugs of abuse to forensic blood samples.

    PubMed

    Moriya, F; Hashimoto, Y

    1996-04-01

    A simple and rapid screening procedure with Triage has been developed to detect 7 classes of drugs of abuse, phencyclidine (PCP), benzodiazepines (BZO), cocaine metabolite (COC), amphetamines (AMP), cannabinoids (THC), opiates (OPI), and barbiturates (BAR), in hemolyzed blood. A clear supernatant was obtained by mixing the blood with sulfosalicylic acid. The supernatant was neutralized with ammonium acetate and then screened using Triage. The lower limits of detection of the Triage screening method for PCP, diazepam, benzolyecgonine, methamphetamine, morphine, 11-nor-delta-9-tetrahydrocannabinol-9-carboxylic acid (THC-COOH), phenobarbital, and secobarbital were 50 ng/mL, 900 ng/mL, 1,000 ng/mL, 600 ng/mL, 900 ng/mL, and 900 ng/mL, respectively. The sensitivity of Triage for THC-COOH in deproteinized blood samples was much lower than that in urine samples. No false positive reactions were observed for the 6 classes of the drugs of abuse with the exception of AMP when the blood was decomposed. Phenethylamine, a putrefactive amine, gave positive results for AMP at concentrations over 5,000 ng/mL. The method was applied to 9 hemolyzed blood samples and 3 turbid urine samples from 12 forensic autopsy cases suspected of drug misuse. Among these, 5 were positive for AMP, 1 for OPI, and 4 for BAR. The presence of methamphetamine is only one of the 5, codeine in 1, and phenobarbital in 4 was confirmed by gas chromatography. All 4 samples which were false positive for AMP contained phenethylamine at relatively high concentrations because of moderate to heavy putrefaction. This method, although disadvantageous to test for AMP and THC, is helpful for the forensic toxicologist because any kind of bloody fluid can be tested rapidly with Triage to detect toxic levels of PCP, BZO, COC, OPI, and BAR.

  11. Barrier screens: a method to sample blood-fed and host-seeking exophilic mosquitoes

    PubMed Central

    2013-01-01

    Background Determining the proportion of blood meals on humans by outdoor-feeding and resting mosquitoes is challenging. This is largely due to the difficulty of finding an adequate and unbiased sample of resting, engorged mosquitoes to enable the identification of host blood meal sources. This is particularly difficult in the south-west Pacific countries of Indonesia, the Solomon Islands and Papua New Guinea where thick vegetation constitutes the primary resting sites for the exophilic mosquitoes that are the primary malaria and filariasis vectors. Methods Barrier screens of shade-cloth netting attached to bamboo poles were constructed between villages and likely areas where mosquitoes might seek blood meals or rest. Flying mosquitoes, obstructed by the barrier screens, would temporarily stop and could then be captured by aspiration at hourly intervals throughout the night. Results In the three countries where this method was evaluated, blood-fed females of Anopheles farauti, Anopheles bancroftii, Anopheles longirostris, Anopheles sundaicus, Anopheles vagus, Anopheles kochi, Anopheles annularis, Anopheles tessellatus, Culex vishnui, Culex quinquefasciatus and Mansonia spp were collected while resting on the barrier screens. In addition, female Anopheles punctulatus and Armigeres spp as well as male An. farauti, Cx. vishnui, Cx. quinquefasciatus and Aedes species were similarly captured. Conclusions Building barrier screens as temporary resting sites in areas where mosquitoes were likely to fly was an extremely time-effective method for collecting an unbiased representative sample of engorged mosquitoes for determining the human blood index. PMID:23379959

  12. Barrier screens: a method to sample blood-fed and host-seeking exophilic mosquitoes.

    PubMed

    Burkot, Thomas R; Russell, Tanya L; Reimer, Lisa J; Bugoro, Hugo; Beebe, Nigel W; Cooper, Robert D; Sukawati, Supraman; Collins, Frank H; Lobo, Neil F

    2013-02-05

    Determining the proportion of blood meals on humans by outdoor-feeding and resting mosquitoes is challenging. This is largely due to the difficulty of finding an adequate and unbiased sample of resting, engorged mosquitoes to enable the identification of host blood meal sources. This is particularly difficult in the south-west Pacific countries of Indonesia, the Solomon Islands and Papua New Guinea where thick vegetation constitutes the primary resting sites for the exophilic mosquitoes that are the primary malaria and filariasis vectors. Barrier screens of shade-cloth netting attached to bamboo poles were constructed between villages and likely areas where mosquitoes might seek blood meals or rest. Flying mosquitoes, obstructed by the barrier screens, would temporarily stop and could then be captured by aspiration at hourly intervals throughout the night. In the three countries where this method was evaluated, blood-fed females of Anopheles farauti, Anopheles bancroftii, Anopheles longirostris, Anopheles sundaicus, Anopheles vagus, Anopheles kochi, Anopheles annularis, Anopheles tessellatus, Culex vishnui, Culex quinquefasciatus and Mansonia spp were collected while resting on the barrier screens. In addition, female Anopheles punctulatus and Armigeres spp as well as male An. farauti, Cx. vishnui, Cx. quinquefasciatus and Aedes species were similarly captured. Building barrier screens as temporary resting sites in areas where mosquitoes were likely to fly was an extremely time-effective method for collecting an unbiased representative sample of engorged mosquitoes for determining the human blood index.

  13. Differences between blood donors and a population sample: implications for case–control studies

    PubMed Central

    Golding, Jean; Northstone, Kate; Miller, Laura L; Davey Smith, George; Pembrey, Marcus

    2013-01-01

    Background Selecting appropriate controls for studies of genetic variation in case series is important. The two major candidates involve the use of blood donors or a random sample of the population. Methods We compare and contrast the two different populations of controls for studies of genetic variation using data from parents enrolled in the Avon Longitudinal Study of Parents and Children (ALSPAC). In addition we compute different biases using a series of hypothetical assumptions. Results The study subjects who had been blood donors differed markedly from the general population in social, health-related, anthropometric, and personality-related variables. Using theoretical examples, we show that blood donors are a poor control group for non-genetic studies of diseases related to environmentally, behaviourally, or socially patterned exposures. However, we show that if blood donors are used as controls in genetic studies, these factors are unlikely to make a major difference in detecting true associations with relatively rare disorders (cumulative incidence through life of <10%). Nevertheless, for more common disorders, the reduction in accuracy resulting from the inclusion in any control population of individuals who have or will develop the disease in question can create a greater bias than can socially patterned factors. Conclusions Information about the medical history of a control and the parents of the control (as a proxy for whether the control will develop the disease) is more important with regard to the choice of controls than whether the controls are a random population sample or blood donors. PMID:23825379

  14. Direct RNA-based detection of CTX-M β-lactamases in human blood samples.

    PubMed

    Stein, Claudia; Makarewicz, Oliwia; Pfeifer, Yvonne; Brandt, Christian; Pletz, Mathias W

    2015-05-01

    Bloodstream infections with ESBL-producers are associated with increased mortality, which is due to delayed appropriate treatment resulting in clinical failure. Current routine diagnostics for detection of bloodstream infections consists of blood culture followed by species identification and susceptibility testing. In attempts to improve and accelerate diagnostic procedures, PCR-based methods have been developed. These methods focus on species identification covering only a limited number of ESBL coding genes. Therefore, they fail to cover the steadily further evolving genetic diversity of clinically relevant β-lactamases. We have recently designed a fast and novel RNA targeting method to detect and specify CTX-M alleles from bacterial cultures, based on an amplification-pyrosequencing approach. We further developed this assay towards a diagnostic tool for clinical use and evaluated its sensitivity and specificity when applied directly to human blood samples. An optimized protocol for mRNA isolation allows detection of specific CTX-M groups from as little as 100 CFU/mL blood via reverse transcription, amplification, and pyrosequencing directly from human EDTA blood samples as well as from pre-incubated human blood cultures with a turnaround time for test results of <7 h. Copyright © 2015 Elsevier GmbH. All rights reserved.

  15. Attic dust and human blood samples collected near a former wood treatment facility.

    PubMed

    Hensley, A R; Scott, A; Rosenfeld, P E; Clark, J J J

    2007-10-01

    A wood treatment facility operating in southern Alabama released dioxins and other hazardous substances into the surrounding community over a period of approximately 35 years. The facility used a variety of chemical insecticides including pentachlorophenol (PCP), chromated copper arsenate (CCA), and creosote (which contains polycyclic aromatic hydrocarbons (PAHs)) to treat wood. The health risks associated with the released contaminants are numerous and significant. To evaluate the historic exposure to the contaminants from the facility, blood samples and health surveys were collected from 21 current and past residents of the adjacent, isolated community and analyzed for concentrations of polychlorinated dibenzo-p-dioxins (dioxins) and polychlorinated dibenzofurans (furans). In addition, attic dust sampling was performed in 11 buildings located within a 1-mile radius of the former wood treatment facility. The average total dioxin toxic equivalent (TEQ) concentration in the residents' blood samples was 36.6 pg/g lipids. In the attic dust, the average total dioxin TEQ concentration, benzo[a]pyrene (PAH) TEQ concentration, and arsenic concentration were 145 ng/kg, 0.98 and 29.8 mg/kg, respectively. The concentrations of dioxins measured in the blood samples exceed the 90th percentile total dioxin levels found in the general US adult population. Concentrations of dioxin, arsenic, and PAHs found in the attic samples exceeded the US Environmental Protection Agency (USEPA) Region 4 soil exposure cancer risk preliminary remediation goal (PRG) values. These findings indicate a very significant potential for related health effects in these communities.

  16. Analysis of whole blood samples with low gas flow inductively coupled plasma-optical emission spectrometry.

    PubMed

    Nowak, Sascha; Künnemeyer, Jens; Terborg, Lydia; Trümpler, Stefan; Günsel, Andreas; Wiesmüller, Gerhard A; Karst, Uwe; Buscher, Wolfgang

    2015-01-01

    Low gas flow ICP-OES with a total argon consumption below 0.7 L/min is introduced for the analysis of trace elements in blood samples to investigate the influence of samples containing an organic solvent in a demanding matrix on the performance of this plasma for the first time. Therefore, gadolinium was determined in human plasma samples and mercury in red blood cells, human plasma, and precipitated plasma protein fraction. Limits of detection (LOD) were determined to be in the low microgram per liter range for the analytes and the accuracy of the method was assessed by comparison with a conventional Fassel-type torch-based ICP-OES. It was proven that the low gas flow ICP-OES leads to comparable results with the instrument based on the Fassel-type torch.

  17. Global oxygen extraction fraction by blood sampling to anticipate cerebral hyperperfusion phenomenon after carotid artery stenting.

    PubMed

    Iwata, Tomonori; Mori, Takahisa; Miyazaki, Yuichi; Tanno, Yuhei; Kasakura, Shigen; Aoyagi, Yoshinori

    2014-11-01

    Cerebral hyperperfusion syndrome sometimes occurs after carotid revascularization in patients with severe hemodynamic failure. To prevent cerebral hyperperfusion syndrome, cerebral hyperperfusion phenomenon (CHP) must be detected early. Single-photon emission computed tomography (SPECT) is useful for detecting CHP, but it is impractical on a daily basis. A tool with high availability to find CHP is desired. To investigate whether global oxygen extraction fraction (OEF) by a blood sampling method is useful for indicating CHP after carotid artery stenting (CAS). When patients underwent elective CAS from September 2010 to August 2012, we performed blood sampling for OEF calculation and SPECT before and immediately after elective CAS. Data were collected prospectively. OEF was calculated from the cerebral arteriovenous oxygen difference. Cerebral blood flow was measured in the affected middle cerebral artery (MCA) territory and in the ipsilateral cerebellum by SPECT. The ratio of MCA to cerebellar activity was defined as cerebral blood flow in the affected MCA territory divided by cerebral blood flow in the ipsilateral cerebellar hemisphere. Probable CHP was defined as ≥10% increase in the ratio of MCA to cerebellar activity after CAS. The relationship between peri-CAS OEF and probable CHP was evaluated. Of the 96 patients enrolled, 92 patients were analyzed. Probable CHP occurred in 17 patients. Post-CAS OEF was related to probable CHP (P < .01), but pre-CAS OEF was not. The receiver-operating characteristic curve showed that the cutoff value was 45% for probable CHP (P < .001). An increase in blood sampling OEF immediately after CAS was related to probable CHP; then the oxygen demand should be reduced.

  18. Polypropylene fibre web, a new matrix for sampling blood for immunodiagnosis of schistosomiasis.

    PubMed

    Jamaly, S; Chihani, T; Deelder, A M; Gabone, R; Nilsson, L A; Ouchterlony, O

    1997-01-01

    Blood sampling on filter paper is widely used for immunodiagnostic and epidemiological purposes. However, elution of conventional filter papers impregnated with sera containing Schistosoma mansoni circulating anodic antigen (CAA) recovered only a small fraction of the antigen, thereby reducing the sensitivity of the assay. Polypropylene-based non-woven fibre web is a new sampling material with a low density of fibres and with a small surface area of contact. When it was impregnated with serum containing CAA, approximately 90% of the antigen could be extracted. The yield of antibodies against S. mansoni from the new sampling material did not differ from that from conventional filter papers.

  19. Prediction of Arterial Blood pH and Partial Pressure of Carbon dioxide from Venous Blood Samples in Patients Receiving Mechanical Ventilation.

    PubMed

    Tavakol, Kamran; Ghahramanpoori, Bahareh; Fararouei, Mohammad

    2013-07-01

    Substitution of arterial with venous blood samples to estimate blood gas status is highly preferable due to practical and safety concerns. Numerous studies support the substitution of arterial by venous blood samples, reporting strong correlations between arterial and venous values. This study further investigated the predictive ability of venous blood samples for arterial Acid-Base Balance (pH) and pressure of carbon dioxide (pCO2). Participants were 51 post-brain surgery patients receiving mechanical ventilation, who had blood samples taken simultaneously from radial artery of the wrist and elbow vein. Results showed significant associations between arterial and venous pH and pCO2. However, the variation of regression residuals was not homogenous, and the regression line did not fit properly to the data, indicating that simple linear regression is sub-optimal for prediction of arterial pH and pCO2 by venous blood sample. Although highly significant correlations were found between arterial and venous blood pH and pCO2, the results did not support the reliability of prediction of arterial blood pH and pCO2 by venous blood samples across a range of concentrations.

  20. Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples

    PubMed Central

    Gomes, Cláudia; Martinez-Puchol, Sandra; Pons, Maria J.; Bazán, Jorge; Tinco, Carmen; del Valle, Juana; Ruiz, Joaquim

    2016-01-01

    Background The lack of an effective diagnostic tool for Carrion’s disease leads to misdiagnosis, wrong treatments and perpetuation of asymptomatic carriers living in endemic areas. Conventional PCR approaches have been reported as a diagnostic technique. However, the detection limit of these techniques is not clear as well as if its usefulness in low bacteriemia cases. The aim of this study was to evaluate the detection limit of 3 PCR approaches. Methodology/Principal Findings We determined the detection limit of 3 different PCR approaches: Bartonella-specific 16S rRNA, fla and its genes. We also evaluated the viability of dry blood spots to be used as a sample transport system. Our results show that 16S rRNA PCR is the approach with a lowest detection limit, 5 CFU/μL, and thus, the best diagnostic PCR tool studied. Dry blood spots diminish the sensitivity of the assay. Conclusions/Significance From the tested PCRs, the 16S rRNA PCR-approach is the best to be used in the direct blood detection of acute cases of Carrion’s disease. However its use in samples from dry blood spots results in easier management of transport samples in rural areas, a slight decrease in the sensitivity was observed. The usefulness to detect by PCR the presence of low-bacteriemic or asymptomatic carriers is doubtful, showing the need to search for new more sensible techniques. PMID:26959642

  1. Raman Spectroscopy: A New Proposal for the Detection of Leukemia Using Blood Samples

    SciTech Connect

    Martinez-Espinosa, J. C.; Gonzalez-Solis, J. L.; Miranda-Beltran, M. L.; Soria-Fregoso, C.; Medina-Valtierra, J.; Sanchez-Gomez, R.

    2008-08-11

    The use of Raman spectroscopy to analyze blood biochemistry and hence distinguish between normal and abnormal blood was investigated. The blood samples were obtained from 6 patients who were clinically diagnosed with leukemia and 6 healthy volunteer. The imprint was put under the microscope and several points were chosen for Raman measurement. All spectra were collected at confocal Raman micro-spectroscopy (Renishaw) with NIR 830 nm laser. It is shown that the serum samples from patients with leukemia and from the control group can be discriminated when the multivariate statistical methods of principal component analysis (PCA) and linear discriminated analysis (LDA) is applied to their Raman spectra. The ratios of some band intensities were analyzed and some band ratios were significant and corresponded to proteins, phospholipids, and polysaccharides. In addition, currently the degree of damage to the bone marrow is estimated through biopsies and therefore it is a very procedure painful. The preliminary results suggest that Raman spectroscopy could be a new technique to study the bone marrow using just blood samples.

  2. Raman Spectroscopy: A New Proposal for the Detection of Leukemia Using Blood Samples

    NASA Astrophysics Data System (ADS)

    Martínez-Espinosa, J. C.; González-Solís, J. L.; Frausto-Reyes, C.; Miranda-Beltrán, M. L.; Soria-Fregoso, C.; Medina-Valtierra, J.; Sánchez-Gómez, R.

    2008-08-01

    The use of Raman spectroscopy to analyze blood biochemistry and hence distinguish between normal and abnormal blood was investigated. The blood samples were obtained from 6 patients who were clinically diagnosed with leukemia and 6 healthy volunteer. The imprint was put under the microscope and several points were chosen for Raman measurement. All spectra were collected at confocal Raman micro-spectroscopy (Renishaw) with NIR 830 nm laser. It is shown that the serum samples from patients with leukemia and from the control group can be discriminated when the multivariate statistical methods of principal component analysis (PCA) and linear discriminated analysis (LDA) is applied to their Raman spectra. The ratios of some band intensities were analyzed and some band ratios were significant and corresponded to proteins, phospholipids, and polysaccharides. In addition, currently the degree of damage to the bone marrow is estimated through biopsies and therefore it is a very procedure painful. The preliminary results suggest that Raman spectroscopy could be a new technique to study the bone marrow using just blood samples.

  3. Detection of Leukemia with Blood Samples Using Raman Spectroscopy and Multivariate Analysis

    NASA Astrophysics Data System (ADS)

    Martínez-Espinosa, J. C.; González-Solís, J. L.; Frausto-Reyes, C.; Miranda-Beltrán, M. L.; Soria-Fregoso, C.; Medina-Valtierra, J.

    2009-06-01

    The use of Raman spectroscopy to analyze blood biochemistry and hence distinguish between normal and abnormal blood was investigated. Blood samples were obtained from 6 patients who were clinically diagnosed with leukemia and 6 healthy volunteers. The imprint was put under the microscope and several points were chosen for Raman measurement. All the spectra were collected by a confocal Raman micro-spectroscopy (Renishaw) with a NIR 830 nm laser. It is shown that the serum samples from patients with leukemia and from the control group can be discriminated when the multivariate statistical methods of principal component analysis (PCA) and linear discriminated analysis (LDA) are applied to their Raman spectra. The ratios of some band intensities were analyzed and some band ratios were significant and corresponded to proteins, phospholipids, and polysaccharides. The preliminary results suggest that Raman Spectroscopy could be a new technique to study the degree of damage to the bone marrow using just blood samples instead of biopsies, treatment very painful for patients.

  4. Severe anaemia associated with Plasmodium falciparum infection in children: consequences for additional blood sampling for research.

    PubMed

    Kuijpers, Laura Maria Francisca; Maltha, Jessica; Guiraud, Issa; Kaboré, Bérenger; Lompo, Palpouguini; Devlieger, Hugo; Van Geet, Chris; Tinto, Halidou; Jacobs, Jan

    2016-06-02

    Plasmodium falciparum infection may cause severe anaemia, particularly in children. When planning a diagnostic study on children suspected of severe malaria in sub-Saharan Africa, it was questioned how much blood could be safely sampled; intended blood volumes (blood cultures and EDTA blood) were 6 mL (children aged <6 years) and 10 mL (6-12 years). A previous review [Bull World Health Organ. 89: 46-53. 2011] recommended not to exceed 3.8 % of total blood volume (TBV). In a simulation exercise using data of children previously enrolled in a study about severe malaria and bacteraemia in Burkina Faso, the impact of this 3.8 % safety guideline was evaluated. For a total of 666 children aged >2 months to <12 years, data of age, weight and haemoglobin value (Hb) were available. For each child, the estimated TBV (TBVe) (mL) was calculated by multiplying the body weight (kg) by the factor 80 (ml/kg). Next, TBVe was corrected for the degree of anaemia to obtain the functional TBV (TBVf). The correction factor consisted of the rate 'Hb of the child divided by the reference Hb'; both the lowest ('best case') and highest ('worst case') reference Hb values were used. Next, the exact volume that a 3.8 % proportion of this TBVf would present was calculated and this volume was compared to the blood volumes that were intended to be sampled. When applied to the Burkina Faso cohort, the simulation exercise pointed out that in 5.3 % (best case) and 11.4 % (worst case) of children the blood volume intended to be sampled would exceed the volume as defined by the 3.8 % safety guideline. Highest proportions would be in the age groups 2-6 months (19.0 %; worst scenario) and 6 months-2 years (15.7 %; worst case scenario). A positive rapid diagnostic test for P. falciparum was associated with an increased risk of violating the safety guideline in the worst case scenario (p = 0.016). Blood sampling in children for research in P. falciparum endemic settings may easily violate

  5. Genetic Characterization of Atypical Mansonella (Mansonella) ozzardi Microfilariae in Human Blood Samples from Northeastern Peru

    PubMed Central

    Marcos, Luis A.; Arrospide, Nancy; Recuenco, Sergio; Cabezas, Cesar; Weil, Gary J.; Fischer, Peter U.

    2012-01-01

    DNA sequence comparisons are useful for characterizing proposed new parasite species or strains. Microfilariae with an atypical arrangement of nuclei behind the cephalic space have been recently described in human blood samples from the Amazon region of Peru. Three blood specimens containing atypical microfilariae were genetically characterized using three DNA markers (5S ribosomal DNA, 12S ribosomal DNA, and cytochrome oxidase I). All atypical microfilariae were clustered into the Mansonella group and indistinguishable from M. ozzardi based on these DNA markers. PMID:22826497

  6. Confocal backscattering-based detection of leukemic cells in flowing blood samples.

    PubMed

    Greiner, Cherry; Hunter, Martin; Rius, Francisca; Huang, Peter; Georgakoudi, Irene

    2011-10-01

    The prognostic value of assessing minimal residual disease (MRD) in leukemia has been established with advancements in flow cytometry and PCR. Nonetheless, these techniques are limited by high equipment costs, complex, and costly cell processing and the need for highly trained personnel. Here, we demonstrate the potential of exploiting differences in the relative intensities of backscattered light at three wavelengths to detect the presence of leukemic cells in samples containing varying mixtures of white blood cells (WBCs) and leukemic cells flowing through microfluidic channels. Using 405, 488, and 633 nm illumination, we identify distinct light scattering intensity distributions for Nalm-6 leukemic cells, normal mononuclear (PBMC) and polymorphonuclear (PMN) white blood cells and red blood cells. We exploit these differences to develop cell classification algorithms, whose performance is evaluated based on simultaneous acquisition of light scattering and fluorescence flow cytometry data. When this algorithm is used prospectively for the analysis of samples consisting of mixtures of PBMCs and leukemic cells, we achieve an average specificity and sensitivity of leukemic cell detection of 99.6 and 45.2%, respectively. When we consider samples that include leukemic cells along with PMNs and PBMCs, which can be acquired using a simple red blood cell lysis step following venipuncture, the specificity and sensitivity of the approach decreases to 91.6 and 39.5%, respectively. On the basis of the performance of these algorithms, we estimate that 42 or 71 μL of blood would be adequate to confirm the presence of leukemia at an 80% power level in samples containing 0.01% leukemia to either PBMCs or PBMCs and PMNs, respectively. Therefore, light scattering-based flow cytometry in a microfluidic platform could provide a low cost, highly portable, minimally invasive approach for detection and monitoring of leukemic patients. This could offer significant improvements

  7. Critical review and meta-analysis of spurious hemolysis in blood samples collected from intravenous catheters

    PubMed Central

    Lippi, Giuseppe; Cervellin, Gianfranco; Mattiuzzi, Camilla

    2013-01-01

    Background: A number of preanalytical activities strongly influence sample quality, especially those related to sample collection. Since blood drawing through intravenous catheters is reported as a potential source of erythrocyte injury, we performed a critical review and meta-analysis about the risk of catheter-related hemolysis. Materials and methods: We performed a systematic search on PubMed, Web of Science and Scopus to estimate the risk of spurious hemolysis in blood samples collected from intravenous catheters. A meta-analysis with calculation of Odds ratio (OR) and Relative risk (RR) along with 95% Confidence interval (95% CI) was carried out using random effect mode. Results: Fifteen articles including 17 studies were finally selected. The total number of patients was 14,796 in 13 studies assessing catheter and evacuated tubes versus straight needle and evacuated tubes, and 1251 in 4 studies assessing catheter and evacuated tubes versus catheter and manual aspiration. A significant risk of hemolysis was found in studies assessing catheter and evacuated tubes versus straight needle and evacuated tubes (random effect OR 3.4; 95% CI = 2.9–3.9 and random effect RR 1.07; 95% CI = 1.06–1.08), as well as in studies assessing catheter and evacuated tubes versus catheter and manual aspiration of blood (OR 3.7; 95% CI = 2.7–5.1 and RR 1.32; 95% CI = 1.24–1.40). Conclusions: Sample collection through intravenous catheters is associated with significant higher risk of spurious hemolysis as compared with standard blood drawn by straight needle, and this risk is further amplified when intravenous catheter are associated with primary evacuated blood tubes as compared with manual aspiration. PMID:23894864

  8. Critical review and meta-analysis of spurious hemolysis in blood samples collected from intravenous catheters.

    PubMed

    Lippi, Giuseppe; Cervellin, Gianfranco; Mattiuzzi, Camilla

    2013-01-01

    A number of preanalytical activities strongly influence sample quality, especially those related to sample collection. Since blood drawing through intravenous catheters is reported as a potential source of erythrocyte injury, we performed a critical review and meta-analysis about the risk of catheter-related hemolysis. We performed a systematic search on PubMed, Web of Science and Scopus to estimate the risk of spurious hemolysis in blood samples collected from intravenous catheters. A meta-analysis with calculation of Odds ratio (OR) and Relative risk (RR) along with 95% Confidence interval (95% CI) was carried out using random effect mode. Fifteen articles including 17 studies were finally selected. The total number of patients was 14,796 in 13 studies assessing catheter and evacuated tubes versus straight needle and evacuated tubes, and 1251 in 4 studies assessing catheter and evacuated tubes versus catheter and manual aspiration. A significant risk of hemolysis was found in studies assessing catheter and evacuated tubes versus straight needle and evacuated tubes (random effect OR 3.4; 95% CI = 2.9-3.9 and random effect RR 1.07; 95% CI = 1.06-1.08), as well as in studies assessing catheter and evacuated tubes versus catheter and manual aspiration of blood (OR 3.7; 95% CI = 2.7-5.1 and RR 1.32; 95% CI = 1.24-1.40). Sample collection through intravenous catheters is associated with significant higher risk of spurious hemolysis as compared with standard blood drawn by straight needle, and this risk is further amplified when intravenous catheter are associated with primary evacuated blood tubes as compared with manual aspiration.

  9. Postmortem tissue samples: an alternative to urine and blood for drug analysis in racehorses.

    PubMed

    Uboh, C E; Rudy, J A; Railing, F A; Enright, J M; Shoemaker, J M; Kahler, M C; Shellenberger, J M; Kemecsei, Z; Das, D N

    1995-09-01

    Although urine is the sample of choice for drug tests in racehorses, it is rarely obtained following the sudden death of a racehorse on the track while racing. The purpose of this study was to demonstrate the significance of postmortem tissue samples as an alternative to urine and blood samples in equine drug analysis following the sudden death of a racehorse on the track while participating in a competitive race. Postmortem tissue samples were frozen (-80 degrees C) until analyzed. A 30-40-g portion of each organ was homogenized in a 0.1 M phosphate buffer (pH 7.4), deproteinized, hydrolyzed with beta-glucuronidase, extracted, and screened by thin-layer chromatography and immunoassay. Samples that initially tested positive for drug(s) were then extracted using high-flow, solid-phase extraction cartridges. The eluates were analyzed by gas chromatography-mass spectrometry. The presence of butorphanol in horses HB355 and CD387, pentobarbital in horse HO940, and ergotamine in horses HO940 and CD387 was detected and confirmed. Thus, in the absence of urine and blood samples following sudden death, postmortem tissue samples are equally useful for forensic toxicological investigations of racehorses.

  10. Evaluation of the effects of insufficient blood volume samples on the performance of blood glucose self-test meters.

    PubMed

    Pfützner, Andreas; Schipper, Christina; Ramljak, Sanja; Flacke, Frank; Sieber, Jochen; Forst, Thomas; Musholt, Petra B

    2013-11-01

    Accuracy of blood glucose readings is (among other things) dependent on the test strip being completely filled with sufficient sample volume. The devices are supposed to display an error message in case of incomplete filling. This laboratory study was performed to test the performance of 31 commercially available devices in case of incomplete strip filling. Samples with two different glucose levels (60-90 and 300-350 mg/dl) were used to generate three different sample volumes: 0.20 µl (too low volume for any device), 0.32 µl (borderline volume), and 1.20 µl (low but supposedly sufficient volume for all devices). After a point-of-care capillary reference measurement (StatStrip, NovaBiomedical), the meter strip was filled (6x) with the respective volume, and the response of the meters (two devices) was documented (72 determinations/meter type). Correct response was defined as either an error message indicating incomplete filling or a correct reading (±20% compared with reference reading). Only five meters showed 100% correct responses [BGStar and iBGStar (both Sanofi), ACCU-CHEK Compact+ and ACCU-CHEK Mobile (both Roche Diagnostics), OneTouch Verio (LifeScan)]. The majority of the meters (17) had up to 10% incorrect reactions [predominantly incorrect readings with sufficient volume; Precision Xceed and Xtra, FreeStyle Lite, and Freedom Lite (all Abbott); GlucoCard+ and GlucoMen GM (both Menarini); Contour, Contour USB, and Breeze2 (all Bayer); OneTouch Ultra Easy, Ultra 2, and Ultra Smart (all LifeScan); Wellion Dialog and Premium (both MedTrust); FineTouch (Terumo); ACCU-CHEK Aviva (Roche); and GlucoTalk (Axis-Shield)]. Ten percent to 20% incorrect reactions were seen with OneTouch Vita (LifeScan), ACCU-CHEK Aviva Nano (Roche), OmniTest+ (BBraun), and AlphaChek+ (Berger Med). More than 20% incorrect reactions were obtained with Pura (Ypsomed), GlucoCard Meter and GlucoMen LX (both Menarini), Elite (Bayer), and MediTouch (Medisana). In summary, partial and

  11. Sample stability for complete blood cell count using the Sysmex XN haematological analyser

    PubMed Central

    Daves, Massimo; Zagler, Elmar M.; Cemin, Roberto; Gnech, Flora; Joos, Alexandra; Platzgummer, Stefan; Lippi, Giuseppe

    2015-01-01

    Background Sample stability is a crucial aspect for the quality of results of a haematology laboratory. This study was conducted to investigate the reliability of haematological testing using Sysmex XN in samples stored for up to 24 h at different temperatures. Materials and methods Haematological tests were performed on whole blood samples collected from 16 ostensibly healthy outpatients immediately after collection and 3 h, 6 h or 24 h afterwards, with triple aliquots kept at room temperature, 4 °C or 37 °C. Results No meaningful bias was observed after 3 h under different storage conditions, except for red blood cell distribution width (RDW) and platelet count (impedance technique, PLT-I) at 37 °C. After 6 h, meaningful bias was observed for mean corpuscular haemoglobin (MCH) and mean corpuscular volume (MCV) at room temperature, red blood cell (RBC) count, mean corpuscular haemoglobin concentration (MCHC), MCH, MCV and PLT-I at 4 °C, and RBC, RDW, MCHC, MCH and PLT-I at 37 °C. After 24 h, a meaningful bias was observed for MCHC, MCV, platelet count (fluorescent technique, PLT-F) and mean platelet volume (MPV) at room temperature, MCHC, MCV, PLT-I and MPV at 4 °C, and all parameters except RBC count and MPV at 37 °C. Discussion Great caution should be observed when analysing results of haematological tests conducted more than 3 h after sample collection. PMID:26057491

  12. LC-MS assay for quantitative determination of cardio glycoside in human blood samples.

    PubMed

    Frommherz, L; Köhler, H; Brinkmann, B; Lehr, M; Beike, J

    2008-03-01

    A method is described for liquid chromatography-mass spectrometry analysis of the cardio glycosides digoxin and digitoxin in biological samples. The method was optimized for use in the forensic field and, therefore, comprises the determination from whole blood and tissue samples. Sample cleanup by solid phase extraction (SPE) on a functionalized polymeric phase was sufficient to limit matrix suppression to <10% for all analytes. Chromatographic separation was achieved using an RP-8 column. Detection of the cardio glycosides was performed with electrospray ionization in the positive mode. The system was run in single ion monitoring mode, measuring the sodium adducts (M + Na)+ of the analyte and of the internal standard, respectively. The method was fully validated for the analysis of blood samples and was also successfully applied in forensic cases. The method was accurate and precise over a linear concentration range up to 50 ng/g blood. Lower limit of quantitation was 0.2 ng/g for digoxin and 2 ng/g for digitoxin, respectively. As deuterated analyte was used as internal standard, we also present a new microwave-enhanced method for the fast preparation of the labelled analyte within 20 min.

  13. Stability of ethyl glucuronide in urine, post-mortem tissue and blood samples.

    PubMed

    Schloegl, Haiko; Dresen, Sebastian; Spaczynski, Karin; Stoertzel, Mylène; Wurst, Friedrich Martin; Weinmann, Wolfgang

    2006-03-01

    The stability of ethyl glucuronide (EtG) under conditions of degradation was examined in urine samples of nine volunteers and in post-mortem tissue (liver, skeletal muscle) and blood taken from seven corpses at autopsies. Analysis was performed via LC-MS/MS. EtG concentrations in urine samples ranged from 2.5 to 296.5 mg/l. When stored at 4 degrees C in airtight test tubes, EtG concentrations remained relatively constant; when stored at room temperature (RT) for 5 weeks in ventilated vials, variations of EtG concentrations ranged from a 30% decrease to an 80% increase, with an average of 37.5% increase. Liver and skeletal muscle tissue of three corpses with positive blood alcohol concentrations (BAC; ranging from 0.106 to 0.183 g%) were stored for 4 weeks and analysed periodically. EtG concentrations decreased 27.7% on average in 4 weeks storage at RT but EtG was still detectable in all samples with initial EtG concentrations higher than 1 mug/g. Blood and liver samples of four corpses with negative BACs were stored at RT after addition of 0.1 g% ethanol, and no new formation of EtG was observed.

  14. Quantitative Analytical Method for the Determination of Biotinidase Activity in Dried Blood Spot Samples.

    PubMed

    Szabó, Eszter; Szatmári, Ildikó; Szőnyi, László; Takáts, Zoltán

    2015-10-20

    Biotinidase activity assay is included in most newborn screening protocols, and the positive results are confirmed by quantitative enzyme activity measurements. In our study, we describe a new quantitative analytical method for the determination of biotinidase activity using the blood sample deposited onto filter paper as the assay medium, by predepositing N-biotinyl-p-aminobenzoic acid onto the standard sample collection paper. The analysis of the assay mixture requires a simple extraction step from a dried blood spot followed by the quantification of product by LC-MS. The method provides a simple and reliable enzyme assay method that enables the rapid diagnosis of biotinidase deficiency (BD). Out of the measured 36 samples, 13 were healthy with lower enzyme activities, 16 were patients with partial BD, and 7 were patients with profound BD with residual activity below 10%. Expression of enzyme activity in percentage of mean activity of negative controls allows comparison of the different techniques. The obtained results are in good agreement with activity data determined from both dried blood spots and serum samples, giving an informative diagnostic value.

  15. Cancer diagnosis by discrimination between normal and malignant human blood samples using attenuated total reflectance-Fourier transform infrared spectroscopy.

    PubMed

    Khanmohammadi, M; Ansari, M A; Garmarudi, A Bagheri; Hassanzadeh, G; Garoosi, G

    2007-09-01

    FTIR spectroscopy is a common technique for cancer diagnosis. Applied tissue samples are heterogeneous and may be damaged in preparation procedures. Easier sampling, more available samples and also easier process with assured results would be interesting. Whole blood samples include all of these qualifications and our hypothesis was the bio-molecular changes in blood which manifest themselves in different optical signatures, detectable by FTIR spectroscopy. Noncancerous blood samples were differentiated from cancerous ones using ATR-FTIR spectroscopy and LDA classification method. Procedure was 100 percent and 90 percent accurate in prediction of cancerous or noncancerous situation for 33 known and 10 unknown samples, respectively.

  16. Pre-analytical effects of blood sampling and handling in quantitative immunoassays for rheumatoid arthritis☆

    PubMed Central

    Zhao, Xiaoyan; Qureshi, Ferhan; Eastman, P. Scott; Manning, William C.; Alexander, Claire; Robinson, William H.; Hesterberg, Lyndal K.

    2012-01-01

    Variability in pre-analytical blood sampling and handling can significantly impact results obtained in quantitative immunoassays. Understanding the impact of these variables is critical for accurate quantification and validation of biomarker measurements. Particularly, in the design and execution of large clinical trials, even small differences in sample processing and handling can have dramatic effects in analytical reliability, results interpretation, trial management and outcome. The effects of two common blood sampling methods (serum vs. plasma) and two widely-used serum handling methods (on the clot with ambient temperature shipping, “traditional”, vs. centrifuged with cold chain shipping, “protocol”) on protein and autoantibody concentrations were examined. Matched serum and plasma samples were collected from 32 rheumatoid arthritis (RA) patients representing a wide range of disease activity status. Additionally, a set of matched serum samples with two sample handling methods was collected. One tube was processed per manufacturer’s instructions and shipped overnight on cold packs (protocol). The matched tube, without prior centrifugation, was simultaneously shipped overnight at ambient temperatures (traditional). Upon delivery, the traditional tube was centrifuged. All samples were subsequently aliquoted and frozen prior to analysis of protein and autoantibody biomarkers. Median correlation between paired serum and plasma across all autoantibody assays was 0.99 (0.98–1.00) with a median % difference of −3.3 (−7.5 to 6.0). In contrast, observed protein biomarker concentrations were significantly affected by sample types, with median correlation of 0.99 (0.33–1.00) and a median % difference of −10 (−55 to 23). When the two serum collection/handling methods were compared, the median correlation between paired samples for autoantibodies was 0.99 (0.91–1.00) with a median difference of 4%. In contrast, significant increases were observed in

  17. DNA profiling: a valuable tool for quality control of sample logistics including occurrences of suspected sample confusion in a blood donation centre.

    PubMed

    Glock, B; Reisacher, R B K; Schöck, M A; Rogado, J; Feinböck, C; Eschner, G; Eder, E; Mayr, W R

    2002-04-01

    A molecular method for analysing whole-blood samples should be established for quality control of plasma sample logistics. DNA profiles of retention samples (plasma) were compared to profiles of recent donations (whole blood). DNA extraction, amplification and detection were performed using the Qiagen DNA Blood Mini kit, the AmpFFISTR Profiler Plus Kit and capillary electrophoresis, respectively. Matched pairs of full profiles were obtained for all samples investigated, therefore no deviation from the standardized procedures was detected. Modified extraction and amplification protocols enabled DNA profiling to be used for the quality control of plasma samples. Hence, DNA profiling can be used in the blood bank as a safe and easy method for quality control of sample logistics.

  18. Distribution of blood types in a sample of 245 New Zealand non-purebred cats.

    PubMed

    Cattin, R P

    2016-05-01

    To determine the distribution of feline blood types in a sample of non-pedigree, domestic cats in New Zealand, whether a difference exists in this distribution between domestic short haired and domestic long haired cats, and between the North and South Islands of New Zealand; and to calculate the risk of a random blood transfusion causing a severe transfusion reaction, and the risk of a random mating producing kittens susceptible to neonatal isoerythrolysis. The results of 245 blood typing tests in non-pedigree cats performed at the New Zealand Veterinary Pathology (NZVP) and Gribbles Veterinary Pathology laboratories between the beginning of 2009 and the end of 2014 were retrospectively collated and analysed. Cats that were identified as domestic short or long haired were included. For the cats tested at Gribbles Veterinary Pathology 62 were from the North Island, and 27 from the South Island. The blood type distribution differed between samples from the two laboratories (p=0.029), but not between domestic short and long haired cats (p=0.50), or between the North and South Islands (p=0.76). Of the 89 cats tested at Gribbles Veterinary Pathology, 70 (79%) were type A, 18 (20%) type B, and 1 (1%) type AB; for NZVP 139/156 (89.1%) cats were type A, 16 (10.3%) type B, and 1 (0.6%) type AB. It was estimated that 18.3-31.9% of random blood transfusions would be at risk of a transfusion reaction, and neonatal isoerythrolysis would be a risk in 9.2-16.1% of random matings between non-pedigree cats. The results from this study suggest that there is a high risk of complications for a random blood transfusion between non-purebred cats in New Zealand. Neonatal isoerythrolysis should be considered an important differential diagnosis in illness or mortality in kittens during the first days of life.

  19. Time impact on non-activated and kaolin-activated blood samples in thromboelastography.

    PubMed

    Durila, Miroslav; Lukáš, Pavel; Bronský, Jiří; Cvachovec, Karel

    2015-04-15

    The correct methodology of thrombelastography might be influenced by elapsing time. In our study we investigated kaolin activated citrated samples together with non-activated citrated samples in relation to the elapsed times of 0, 15 and 30 minutes to compare both methods and to find out if there is an impact of time on results of thrombelastography. Blood samples obtained from 10 healthy volunteers were analyzed after 0, 15 and 30 minutes from sampling with kaolin activation and without activation. Then the results were analysed and compared between the non-activated and the kaolin-activated method. All blood samples became more hypercoagulable with the time elapsing, both in non-activated and kaolin-activated samples and differences between both groups were found statistically and clinically significant after only 0 minutes. The non-activated citrated method seems to be reliable and suitable for thrombelastography in non-emergency cases (planned surgical procedures) when we have time to wait 15-30 minutes to get results. In urgent situations a rapid thrombelastography test should be preferred. Although the kaolin-activated method can also be used, results must be interpreted with caution.

  20. The Use of Dried Blood Spot Sampling in the National Social Life, Health, and Aging Project

    PubMed Central

    McDade, Thomas W.

    2009-01-01

    Objectives This paper describes the methods used for and issues associated with collection and analysis of dried blood spot (DBS) samples for the National Social Life, Health, and Aging Project and provides the basic distributions of the resulting analytes. Methods DBSs from capillary finger sticks were collected by nonmedically trained interviewers from 2,044 individuals, aged 57–85 years. The quality and quantity of DBS samples were evaluated to allow for analysis of interviewer performance. Levels of C-reactive protein, antibodies to the Epstein–Barr virus, hemoglobin, and glycosylated hemoglobin were assayed using various analytic methods. Results Cooperation rate for DBS collection was 84.5%, with 99% of the cards yielding enough sample for at least one analysis. The distribution, mean, and standard deviation of the analytes obtained from DBSs are also presented in this paper. Conclusions The high cooperation rate and quality of the spots collected suggest that the collection of DBSs in population-based research is a feasible and viable alternative to venous blood draws. The relative ease of sample collection, transport, and storage are significant benefits. Care should be taken, however, when comparing results from analysis of DBS samples with those obtained from serum or plasma samples. PMID:19244547

  1. Sample to answer visualization pipeline for low-cost point-of-care blood cell counting

    NASA Astrophysics Data System (ADS)

    Smith, Suzanne; Naidoo, Thegaran; Davies, Emlyn; Fourie, Louis; Nxumalo, Zandile; Swart, Hein; Marais, Philip; Land, Kevin; Roux, Pieter

    2015-03-01

    We present a visualization pipeline from sample to answer for point-of-care blood cell counting applications. Effective and low-cost point-of-care medical diagnostic tests provide developing countries and rural communities with accessible healthcare solutions [1], and can be particularly beneficial for blood cell count tests, which are often the starting point in the process of diagnosing a patient [2]. The initial focus of this work is on total white and red blood cell counts, using a microfluidic cartridge [3] for sample processing. Analysis of the processed samples has been implemented by means of two main optical visualization systems developed in-house: 1) a fluidic operation analysis system using high speed video data to determine volumes, mixing efficiency and flow rates, and 2) a microscopy analysis system to investigate homogeneity and concentration of blood cells. Fluidic parameters were derived from the optical flow [4] as well as color-based segmentation of the different fluids using a hue-saturation-value (HSV) color space. Cell count estimates were obtained using automated microscopy analysis and were compared to a widely accepted manual method for cell counting using a hemocytometer [5]. The results using the first iteration microfluidic device [3] showed that the most simple - and thus low-cost - approach for microfluidic component implementation was not adequate as compared to techniques based on manual cell counting principles. An improved microfluidic design has been developed to incorporate enhanced mixing and metering components, which together with this work provides the foundation on which to successfully implement automated, rapid and low-cost blood cell counting tests.

  2. Assessment of erythrocyte aggregation in whole blood samples by light backscattering: clinical applications

    NASA Astrophysics Data System (ADS)

    Priezzhev, Alexander V.; Firsov, Nikolai N.; Vyshlova, Marina G.; Lademann, Juergen; Richter, Heike; Kiesewetter, Holger; Mueller, Gerhard J.

    1999-05-01

    We report on the results of a collaborative effort made in the field of optical diagnostics of whole blood samples to study the ability of red blood cells to aggregate in a Couette chamber. We studied a possibility to quantitatively measure this ability as a function of the physiological state of blood donors. The aggregometer designed by the Russian coauthors of this paper and described in their earlier publications (see e.g. Proc SPIE 1884, 2100, 2678, 2982) was extensively used in the experiments performed in the Rheumatology Institute in Moscow and in the Charite Clinic in Berlin. The following parameters were measured: two characteristic times of RBC aggregation and the average spontaneous aggregation rate in the state of stasis, the average hydrodynamic strength of all aggregates and that of the largest aggregates. Different algorithms of the remission signal processing for the quantitative evaluation of the above parameters were compared. Reproducible alterations of the parameters from their normal values were obtained for blood samples from individuals suffering auto-immune disease and diabetes. Statistical data is reported proving high efficiency of the technique for the diagnostics of rheological disorders. Basing on these data the quantitative criteria of the heaviness of hemorheological state of the patients are proposed that are important for choosing specific therapies for which the patient is minimally resistant.

  3. Inactivation of human immunodeficiency virus type 1 in blood samples stored as high-salt lysates.

    PubMed

    Zolg, J W; Lanciotti, R S; Wendlinger, M; Meyer, W A

    1990-09-01

    Blood samples to be tested for the presence of parasite DNA by using specific DNA probes are routinely stored in our laboratory as high-salt lysates (HSL). To safeguard against the risk of accidental infection with etiological agents such as the human immunodeficiency virus type 1 (HIV-1) while manipulating large numbers of blood samples in preparation for DNA probing, we determined the residual infectivity of HIV-1 after exposure to HSL components. Both high-titer virus stocks or provirus-carrying cells, suspended either in tissue culture medium or freshly drawn blood, were completely inactivated upon contact with the HSL components. This was verified by the absence of any detectable HIV-1-specific antigen in the supernatants of long-term cultures and the absence of virus-specific DNA fragments after amplification by polymerase chain reaction with DNA from such cultures as target DNA. These results support the conclusion that the virus is in fact completely inactivated by contact with the HSL components, rendering blood specimens stored as HSL noninfectious in regard to HIV-1.

  4. Microchip Module for Blood Sample Preparation and Nucleic Acid Amplification Reactions

    PubMed Central

    Yuen, Po Ki; Kricka, Larry J.; Fortina, Paolo; Panaro, Nicholas J.; Sakazume, Taku; Wilding, Peter

    2001-01-01

    A computer numerical control-machined plexiglas-based microchip module was designed and constructed for the integration of blood sample preparation and nucleic acid amplification reactions. The microchip module is comprised of a custom-made heater-cooler for thermal cycling, a series of 254 μm × 254 μm microchannels for transporting human whole blood and reagents in and out of an 8–9 μL dual-purpose (cell isolation and PCR) glass-silicon microchip. White blood cells were first isolated from a small volume of human whole blood (<3 μL) in an integrated cell isolation–PCR microchip containing a series of 3.5-μm feature-sized “weir-type” filters, formed by an etched silicon dam spanning the flow chamber. A genomic target, a region in the human coagulation Factor V gene (226-bp), was subsequently directly amplified by microchip-based PCR on DNA released from white blood cells isolated on the filter section of the microchip mounted onto the microchip module. The microchip module provides a convenient means to simplify nucleic acid analyses by integrating two key steps in genetic testing procedures, cell isolation and PCR and promises to be adaptable for additional types of integrated assays. PMID:11230164

  5. Microchip module for blood sample preparation and nucleic acid amplification reactions.

    PubMed

    Yuen, P K; Kricka, L J; Fortina, P; Panaro, N J; Sakazume, T; Wilding, P

    2001-03-01

    A computer numerical control-machined plexiglas-based microchip module was designed and constructed for the integration of blood sample preparation and nucleic acid amplification reactions. The microchip module is comprised of a custom-made heater-cooler for thermal cycling, a series of 254 microm x 254 microm microchannels for transporting human whole blood and reagents in and out of an 8--9 microL dual-purpose (cell isolation and PCR) glass-silicon microchip. White blood cells were first isolated from a small volume of human whole blood (<3 microL) in an integrated cell isolation--PCR microchip containing a series of 3.5-microm feature-sized "weir-type" filters, formed by an etched silicon dam spanning the flow chamber. A genomic target, a region in the human coagulation Factor V gene (226-bp), was subsequently directly amplified by microchip-based PCR on DNA released from white blood cells isolated on the filter section of the microchip mounted onto the microchip module. The microchip module provides a convenient means to simplify nucleic acid analyses by integrating two key steps in genetic testing procedures, cell isolation and PCR and promises to be adaptable for additional types of integrated assays.

  6. Surgical vascular access in the porcine model for long-term repeated blood sampling.

    PubMed

    Lombardo, Carlo; Damiano, Giuseppe; Cassata, Giovanni; Palumbo, Vincenzo Davide; Cacciabaudo, Francesco; Spinelii, Gabriele; Calvagna, Cristiano; Gioviale, Maria Concetta; Maione, Caro-lina; Lo Monte, Attilio Ignazio

    2010-09-01

    A simple technique for implanting a long-term jugular catheter in piglets under general anesthesia is described. We report our experience in 10 young female pigs with a body weight of 20-30 Kg. The surgical procedure involves implantation of a jugular central venous catheter (11Fr polyurethane) tunneled in the subcutaneous fat layer of the neck. This procedure may be performed in about 15 minutes. The maintenance of the catheter is described which allows several daily blood samples to be taken. This procedure reduces both the stress in piglets and the chance of catheter dislodgement due to the animals scratching or rubbing. Blood sampling can be easily performed with a low incidence of infection or thrombosis.

  7. Active tracking of rejected dried blood samples in a large program in Nigeria

    PubMed Central

    Inalegwu, Auchi; Phillips, Sunny; Datir, Rawlings; Chime, Christopher; Ozumba, Petronilla; Peters, Samuel; Ogbanufe, Obinna; Mensah, Charles; Abimiku, Alash’Le; Dakum, Patrick; Ndembi, Nicaise

    2016-01-01

    AIM: To study the impact of rejection at different levels of health care by retrospectively reviewing records of dried blood spot samples received at the molecular laboratory for human immunodeficiency virus (HIV) early infant diagnosis (EID) between January 2008 and December 2012. METHODS: The specimen rejection rate, reasons for rejection and the impact of rejection at different levels of health care was examined. The extracted data were cleaned and checked for consistency and then de-duplicated using the unique patient and clinic identifiers. The cleaned data were ciphered and exported to SPSS version 19 (SPSS 2010 IBM Corp, New York, United States) for statistical analyses. RESULTS: Sample rejection rate of 2.4% (n = 786/32552) and repeat rate of 8.8% (n = 69/786) were established. The mean age of infants presenting for first HIV molecular test among accepted valid samples was 17.83 wk (95%CI: 17.65-18.01) vs 20.30 wk (95%CI: 16.53-24.06) for repeated samples. HIV infection rate was 9.8% vs 15.9% for accepted and repeated samples. Compared to tertiary healthcare clinics, secondary and primary clinics had two-fold and three-fold higher likelihood of sample rejection, respectively (P < 0.05). We observed a significant increase in sample rejection rate with increasing number of EID clinics (r = 0.893, P = 0.041). The major reasons for rejection were improper sample collection (26.3%), improper labeling (16.4%) and insufficient blood (14.8%). CONCLUSION: Programs should monitor pre-analytical variables and incorporate continuous quality improvement interventions to reduce errors associated with sample rejection and improve patient retention. PMID:27175352

  8. Characterization of endothelial colony-forming cells from peripheral blood samples of adult horses.

    PubMed

    Salter, Margaret M; Seeto, Wen J; DeWitt, Blake B; Hashimi, Sarah A; Schwartz, Dean D; Lipke, Elizabeth A; Wooldridge, Anne A

    2015-02-01

    To isolate and characterize endothelial colony-forming cells (ECFCs; a subtype of endothelial progenitor cells) from peripheral blood samples of horses. Jugular venous blood samples from 24 adult horses. Blood samples were cultured in endothelial cell growth medium. Isolated ECFCs were characterized by use of functional assays of fluorescence-labeled acetylated low-density lipoprotein (DiI-Ac-LDL) uptake and vascular tubule formation in vitro. Expression of endothelial (CD34, CD105, vascular endothelial growth factor receptor 2, and von Willebrand factor) and hematopoietic (CD14) cell markers was assessed through indirect immunofluorescence assay and flow cytometry. The number of passages before senescence was determined through serial evaluation of DiI-Ac-LDL uptake, vascular tubule formation, and cell doubling rates. Samples from 3 horses produced colonies at 12 ± 2.5 days with characteristic endothelial single layer cobblestone morphology and substantial outgrowth on expansion. Equine ECFCs formed vascular tubules in vitro and had uptake of DiI-Ac-LDL (74.9 ± 14.7% positive cells). Tubule formation and DiI-Ac-LDL uptake diminished by passage 5. Equine ECFCs tested positive for von Willebrand factor, vascular endothelial growth factor receptor 2, CD34, and CD105 with an immunofluorescence assay and for CD14 and CD105 via flow cytometry. ECFCs can be isolated from peripheral blood of horses and have characteristics similar to those described for other species. These cells may have potential therapeutic use in equine diseases associated with ischemia or delayed vascularization.

  9. Fast and Specific Assessment of the Halogenating Peroxidase Activity in Leukocyte-enriched Blood Samples.

    PubMed

    Flemmig, Jörg; Schwarz, Pauline; Bäcker, Ingo; Leichsenring, Anna; Lange, Franziska; Arnhold, Jürgen

    2016-07-28

    In this paper a protocol for the quick and standardized enrichment of leukocytes from small whole blood samples is described. This procedure is based on the hypotonic lysis of erythrocytes and can be applied to human samples as well as to blood of non-human origin. The small initial sample volume of about 50 to 100 µl makes this method applicable to recurrent blood sampling from small laboratory animals. Moreover, leukocyte enrichment is achieved within minutes and with low material efforts regarding chemicals and instrumentation, making this method applicable in multiple laboratory environments. Standardized purification of leukocytes is combined with a highly selective staining method to evaluate halogenating peroxidase activity of the heme peroxidases, myeloperoxidase (MPO) and eosinophil peroxidase (EPO), i.e., the formation of hypochlorous and hypobromous acid (HOCl and HOBr). While MPO is strongly expressed in neutrophils, the most abundant immune cell type in human blood as well as in monocytes, the related enzyme EPO is exclusively expressed in eosinophils. The halogenating activity of these enzymes is addressed by using the almost HOCl- and HOBr-specific dye aminophenyl fluorescein (APF) and the primary peroxidase substrate hydrogen peroxide. Upon subsequent flow cytometry analysis all peroxidase-positive cells (neutrophils, monocytes, eosinophils) are distinguishable and their halogenating peroxidase activity can be quantified. Since APF staining may be combined with the application of cell surface markers, this protocol can be extended to specifically address leukocyte sub-fractions. The method is applicable to detect HOCl and HOBr production both in human and in rodent leukocytes. Given the widely and diversely discussed immunological role of these enzymatic products in chronic inflammatory diseases, this protocol may contribute to a better understanding of the immunological relevance of leukocyte-derived heme peroxidases.

  10. New reference values for routine blood samples and human neutrophilic lipocalin during third-trimester pregnancy.

    PubMed

    Edelstam, G; Löwbeer, C; Kral, G; Gustafsson, S A; Venge, P

    2001-01-01

    Reference values are usually based on blood samples from healthy men or non-pregnant women. Blood samples from pregnant women may be compared with these reference values. Correct references for pregnancy can be extremely important for clinical decisions such as ablatio placentae, appendicitis, premature rupture of membranes and preeclampsia. Previous studies of normal variations during third-trimester pregnancy are incomplete. Blood samples during pregnancy weeks 33, 36 and 39 as well as 1-3 h postpartum were collected from pregnant women with dietary iron supplement and at least one previous pregancy without a history of hypertension or preeclampsia. When the sampled values were compared with the present reference values from men and non-pregnant women, the following differences were found during normal pregnancy: Haemoglobin and ferritin were reduced, CRP was slightly elevated, WBC (white blood cell count) and HNL (human neutrophilic lipocalin) were elevated during pregnancy and significantly increased postpartum. Albumin was reduced. ALT and AST were slightly elevated and GGT was unchanged during pregnancy. ALP, D-dimer and fibrinogen were elevated. Uric acid increased during the third trimester and thrombocyte count decreased. Separate reference values for pregnant women are essential for correct diagnostic decisions during third-trimester pregnancy. Elevated levels of D-dimer do not necessarily indicate ablatio placentae. A diagnosis of progressive preeclampsia cannot be based on increasing uric acid levels and reduced platelet count in a stable clinical condition. HNL signals activation of neutrophilic granulocytes and can thereby offer a helpful tool for diagnosing infection during pregnancy and postpartum.

  11. Children's views on microneedle use as an alternative to blood sampling for patient monitoring.

    PubMed

    Mooney, Karen; McElnay, James C; Donnelly, Ryan F

    2014-10-01

    To explore children's views on microneedle use for this population, particularly as an alternative approach to blood sampling, in monitoring applications, and so, examine the acceptability of this approach to children. Focus groups were conducted with children (aged 10-14 years) in a range of schools across Northern Ireland. Convenience sampling was employed, i.e. children involved in a university-directed community-outreach project (Pharmacists in Schools) were recruited. A total of 86 children participated in 13 focus groups across seven schools in Northern Ireland. A widespread disapproval for blood sampling was evident, with pain, blood and traditional needle visualisation particularly unpopular aspects. In general, microneedles had greater visual acceptability and caused less fear. A patch-based design enabled minimal patient awareness of the monitoring procedure, with personalised designs, e.g. cartoon themes, favoured. Children's concerns included possible allergy and potential inaccuracies with this novel approach; however, many had confidence in the judgement of healthcare professionals if deeming this technique appropriate. They considered paediatric patient education critical for acceptance of this new approach and called for an alternative name, without any reference to 'needles'. The findings presented here support the development of blood-free, minimally invasive techniques and provide an initial indication of microneedle acceptability in children, particularly for monitoring purposes. A proactive response to these unique insights should enable microneedle array design to better meet the needs of this end-user group. Further work in this area is recommended to ascertain the perspectives of a purposive sample of children with chronic conditions who require regular monitoring. © 2013 Royal Pharmaceutical Society.

  12. Optical detection of Trypanosoma cruzi in blood samples for diagnosis purpose

    NASA Astrophysics Data System (ADS)

    Alanis, Elvio; Romero, Graciela; Alvarez, Liliana; Martinez, Carlos C.; Basombrio, Miguel A.

    2004-10-01

    An optical method for detection of Trypanosoma Cruzi (T. cruzi) parasites in blood samples of mice infected with Chagas disease is presented. The method is intended for use in human blood, for diagnosis purposes. A thin layer of blood infected by T. cruzi parasites, in small concentrations, is examined in an interferometric microscope in which the images of the vision field are taken by a CCD camera and temporarily stored in the memory of a host computer. The whole sample is scanned displacing the microscope plate by means of step motors driven by the computer. Several consecutive images of the same field are taken and digitally processed by means of image temporal differentiation in order to detect if a parasite is eventually present in the field. Each field of view is processed in the same fashion, until the full area of the sample is covered or until a parasite is detected, in which case an acoustical warning is activated and the corresponding image is displayed permitting the technician to corroborate the result visually. A discussion of the reliability of the method as well as a comparison with other well established techniques are presented.

  13. Effect of blood sample handling post-collection on Erysipelothrix rhusiopathiae antibody titres.

    PubMed

    Neumann, Eric J; Bonistalli, Kathryn N

    2009-06-01

    A study was conducted to determine the effect of blood sample mishandling on the performance of an enzyme-linked immunosorbent assay for the detection of antibodies against Erysipelothrix rhusiopathiae. Eleven sample maltreatments (storage at -10 degrees C, storage at 4 degrees C, heat treatment of clotted blood, haemolysis, repetitive freeze-thaw cycling, and substitution of plasma in place of serum) were simulated in a laboratory environment and then run concurrently against a gold standard sample (storage at -80 degrees C). The mishandling treatment groups that simulated high levels of haemolysis had significantly lower optical density (OD) readings when compared to the gold standard. However, the magnitude of the effects was relatively small and only samples with OD values close to the cut-off changed state from positive to negative. Heat treatment had a minor, but non-significant, effect on OD values. Findings from this study suggested that immunoglobulin G antibody was stable in the face of most common sample mishandling events.

  14. Targeting prohibited substances in doping control blood samples by means of chromatographic-mass spectrometric methods.

    PubMed

    Thevis, Mario; Thomas, Andreas; Schänzer, Wilhelm

    2013-12-01

    Urine samples have been the predominant matrix for doping controls for several decades. However, owing to the complementary information provided by blood (as well as serum or plasma and dried blood spots (DBS)), the benefits of its analysis have resulted in continuously increasing appreciation by anti-doping authorities. On the one hand, blood samples allow for the detection of various different methods of blood doping and the abuse of erythropoiesis-stimulating agents (ESAs) via the Athlete Biological Passport; on the other hand, targeted and non-targeted drug detection by means of chromatographic-mass spectrometric methods represents an important tool to increase doping control frequencies out-of-competition and to determine drug concentrations particularly in in-competition scenarios. Moreover, blood analysis seldom requires in-depth knowledge of drug metabolism, and the intact substance rather than potentially unknown or assumed metabolic products can be targeted. In this review, the recent developments in human sports drug testing concerning mass spectrometry-based techniques for qualitative and quantitative analyses of therapeutics and emerging drug candidates are summarized and reviewed. The analytical methods include both low and high molecular mass compounds (e.g., anabolic agents, stimulants, metabolic modulators, peptide hormones, and small interfering RNA (siRNA)) determined from serum, plasma, and DBS using state-of-the-art instrumentation such as liquid chromatography (LC)-high resolution/high accuracy (tandem) mass spectrometry (LC-HRMS), LC-low resolution tandem mass spectrometry (LC-MS/MS), and gas chromatography-mass spectrometry (GC-MS).

  15. The effect of sodium fluoride on the stability of cyanide in postmortem blood samples from fire victims.

    PubMed

    McAllister, J L; Roby, R J; Levine, Barry; Purser, David

    2011-06-15

    Assigning a level of significance to cyanide concentrations found in the blood of fire victims is often hampered by the fact that cyanide is inherently unstable in cadavers and in stored blood samples. A few researchers have proposed that sodium fluoride can be used to minimize the instability of cyanide in blood samples; however, controlled studies have not been performed to support validation of this hypothesis. To test the sodium fluoride hypothesis, both treated and control blood samples from 14 autopsied fire victims were tested over a 25-30 day period. A 2% concentration of sodium fluoride was added to the blood samples at the start of testing and the samples were refrigerated between testing intervals. Cyanide concentrations in the treated and control samples were measured between 9 and 11 days post treatment and between 25 and 30 days post treatment. A statistically significant difference was not present between blood cyanide concentrations in treated and control samples between 9 and 11 days. During this time period, although there were small statistically significant increases in both treated and untreated samples the fluctuations were minor. Since the treated and control samples did not exhibit instability between 9 and 11 days, it is not surprising that the sodium fluoride appeared to have no effect. However, a statistically significant difference between blood cyanide concentrations in treated and control samples was observed between 25 and 30 days. Those samples treated with sodium fluoride showed a reduction in blood cyanide variability with virtually no overall change, over a 25-30 day period when compared to control samples, while unconditioned samples showed a significant, average increase of 35%. Based on the findings of this study, it is recommended that 2% sodium fluoride be added to blood samples obtained from fire victims to reduce cyanide instability due to bacteriological activity.

  16. Necessity and risks of arterial blood sampling in healthy volunteer studies.

    PubMed

    Oertel, Bruno Georg; Vermehren, Johannes; Zimmermann, Michael; Huynh, Thomas Tao; Doehring, Alexandra; Ferreiros, Nerea; Senzel, Stephan; Schmitz-Rixen, Thomas; Erbe, Matthias; Geisslinger, Gerd; Harder, Sebastian; Angst, Martin S; Lötsch, Jörn

    2012-10-01

    Arterial blood sampling is necessary when drugs such as the fast-acting opioid analgesic remifentanil exhibit relevant differences between arterial and venous blood concentrations. Arterial cannulation is generally considered to be clinically safe and has thus become a standard procedure in pharmacokinetic-pharmacodynamic assessments. However, rare cases of arterial occlusions have to be considered in risk-benefit assessments of arterial sampling in pharmacokinetic studies, especially when including healthy volunteers. In an actual case, arterial occlusion requiring surgical repair was caused by a factor V Leiden thrombophilia associated genetic variant F5 1691G>A (rs6025) and aggravated by a hypoplastic radial artery. Neither risk factor had been identified prior to enrolment by routine laboratory tests such as the prothrombin time (international normalized ratio), partial thromboplastin time and the clinical Allen's test of arterial function. Re-assessment of the necessity of arterial sampling showed that none of the potential alternatives, target concentrations of computerized infusions or venous concentrations during non-steady-state and steady-state conditions could provide the arterial concentrations. Relying on venous concentrations may result in erroneous pharmacodynamic parameters. Accurate pharmacokinetic-pharmacodynamic studies relying on precisely measured blood concentrations require serial sampling techniques during both steady-state and non-steady-state conditions. However, as illustrated by the presented case, incidents involving the generally safe procedure of arterial sampling are possible, although rare. To further minimize the risks, screening of subjects for prothrombotic risks and careful assessment of the suitability of the artery should be considered in pharmacokinetic studies requiring arterial cannulation.

  17. Dried Blood Spot sampling in psychiatry: Perspectives for improving therapeutic drug monitoring.

    PubMed

    Martial, Lisa C; Aarnoutse, Rob E; Mulder, Martina; Schellekens, Arnt; Brüggemann, Roger J M; Burger, David M; Schene, Aart H; Batalla, Albert

    2017-03-01

    Assessment of drug concentrations is indicated to guide dosing of a selected number of drugs used in psychiatry. Conventionally this is done by vena puncture. Novel sampling strategies such as dried blood spot (DBS) sampling have been developed for various drugs, including antipsychotics, antidepressants and mood-stabilizers. DBS sampling is typically performed by means of a finger prick. This method allows for remote sampling, which means that patients are not required to travel to a health care facility. The number of DBS assays for drugs used in psychiatry has increased over the last decade and includes antidepressants (tricyclic and serotonin and/or norepinephrine reuptake inhibitors), mood stabilizers and first- and second-generation antipsychotics. Available assays often comply with analytical validation criteria but are seldom used in routine clinical care. Little attention has been paid to the clinical validation and implementation processes of home sampling. Ideally, not only medicines but also clinical chemistry parameters should be measured within the same sample. This article reflects on the position of DBS remote sampling in psychiatry and provides insight in the requisites of making such a sampling tool successful. Copyright © 2017 Elsevier B.V. and ECNP. All rights reserved.

  18. Determination of proflavine in rat whole blood without sample pretreatment by laser desorption postionization mass spectrometry.

    PubMed

    Chen, Jiaxin; Hu, Yongjun; Lu, Qiao; Wang, Pengchao; Zhan, Huaqi

    2017-02-10

    A novel pretreatment-free method involving laser desorption postionization (LDPI) coupled with time-of-flight mass spectrometry (MS) was developed for the monitoring of proflavine level in rat whole blood. It comprises a protocol for dosing via intravenous administration and collection of whole blood, followed by direct LDPI-MS analysis without any sample pretreatment. An intense ion signal at m/z 209 was observed from whole blood without any interference signals, except some background signals below m/z 100. The calibration curve was established with use of 9-phenylacridine as the internal standard for proflavine determination from the plotting of the peak ratios of proflavine to the internal standard, with a correlation coefficient (R (2)) greater than 0.99. The limit of detection was estimated to be 0.48 pmol/mm(2) and the quantification range was 0.5-16.5 μg/mL for proflavine. In addition, only a minimal matrix effect was observed, as expected from considerations of the desorption and ionization mechanism. Interday and intraday accuracy and precision were calculated to be within 13% and 82-114%, respectively. Estimated concentrations of proflavine residue in whole blood were also successfully obtained at selected time points after dosing. The proposed method is simple, low cost, and sensitive, and should be seen as a complementary method for monitoring drug levels in blood. Graphical Abstract Monitoring proflavine levels in rat whole blood at different time points using laser desorption postionization mass spectrometry (LDPI-MS).

  19. Comparison of diagnostic tools for the detection of aspergillosis in blood samples of experimentally infected falcons.

    PubMed

    Fischer, D; Van Waeyenberghe, L; Cray, C; Gross, M; Usleber, E; Pasmans, F; Martel, A; Lierz, M

    2014-12-01

    Antemortem diagnosis of avian aspergillosis is very challenging. Diagnostic assays using blood samples would aid in an early and more definitive diagnosis. In the current study, detection of anti-Aspergillus antibodies, Aspergillus antigen, and Aspergillus toxin (fumigaclavine A), protein electrophoresis and measurement of acute-phase protein concentrations were performed on serum of 18 adult and plasma of 21 juvenile gyr-saker hybrid falcons (Falco rusticolus x Falco cherrug). Adult (n = 15) and juvenile (n = 18) falcons were experimentally inoculated with different dosages of the same strain of Aspergillus fumigatus and an additional three falcons from each age group were used as uninfected control animals. Blood samples were collected prior to inoculation and at 28 days postinoculation. Of the 33 inoculated falcons, 16 demonstrated clinical signs (vomiting, greenish urates, dyspnea, ruffled feathers) commonly associated with aspergillosis and in 14 falcons necropsy revealed aspergillosis granulomas confirmed by mycology and histopathology. Positive galactomannan results were rare, with only 3/15 positive samples from adult falcons and none in the juvenile birds. Most of the inoculated falcons showed an increase of serum amyloid A (66.7%) and haptoglobin (70.4%), but fumigaclavine A was not detected in the blood from any of the experimental animals. Elevated antibody indices were detected in 96.7% of the inoculated birds, but also in 66.7% of the controls. Significant decreases in albumin:globulin ratio were obvious in 81.5% of the inoculated birds, including 100% of the birds with granulomas. Blood from falcons with granulomas demonstrated significantly increased concentration values of alpha 2 and β globulins, decreased percentages of prealbumin and albumin, and increased percentages of alpha 2 and β globulins compared to inoculated falcons without granulomas. In conclusion, acute-phase proteins and the electrophoretic profile of birds challenged with A

  20. Array CGH Analysis of Paired Blood and Tumor Samples from Patients with Sporadic Wilms Tumor

    PubMed Central

    del Carmen Crespo, María; Vallespín, Elena; Palomares-Bralo, María; Martin-Arenas, Rubén; Rueda-Arenas, Inmaculada; Silvestre de Faria, Paulo Antonio; García-Miguel, Purificación; Lapunzina, Pablo; Regla Vargas, Fernando; Seuanez, Hector N.; Martínez-Glez, Víctor

    2015-01-01

    Wilms tumor (WT), the most common cancer of the kidney in infants and children, has a complex etiology that is still poorly understood. Identification of genomic copy number variants (CNV) in tumor genomes provides a better understanding of cancer development which may be useful for diagnosis and therapeutic targets. In paired blood and tumor DNA samples from 14 patients with sporadic WT, analyzed by aCGH, 22% of chromosome abnormalities were novel. All constitutional alterations identified in blood were segmental (in 28.6% of patients) and were also present in the paired tumor samples. Two segmental gains (2p21 and 20q13.3) and one loss (19q13.31) present in blood had not been previously described in WT. We also describe, for the first time, a small, constitutive partial gain of 3p22.1 comprising 2 exons of CTNNB1, a gene associated to WT. Among somatic alterations, novel structural chromosomal abnormalities were found, like gain of 19p13.3 and 20p12.3, and losses of 2p16.1-p15, 4q32.5-q35.1, 4q35.2-q28.1 and 19p13.3. Candidate genes included in these regions might be constitutively (SIX3, SALL4) or somatically (NEK1, PIAS4, BMP2) operational in the development and progression of WT. To our knowledge this is the first report of CNV in paired blood and tumor samples in sporadic WT. PMID:26317783

  1. Dried blood spots as a sampling technique for the quantitative determination of guanfacine in clinical studies.

    PubMed

    Li, Yuanyuan; Henion, Jack; Abbott, Richard; Wang, Phillip

    2011-11-01

    Dried blood spot (DBS) technology was evaluated for the quantitative determination of guanfacine in human blood in clinical studies. A very sensitive DBS assay has been developed using HPLC coupled with an AB Sciex 5500 QTRAP® (Applied Biosystems/MDS Sciex, ON, Canada) MS system (LC-MS/MS) with a linear calibration range of 0.05 to 25 ng/ml. High-resolution MS using an Exactive Orbitrap® (ThermoFisher, LLC., CA, USA) was compared with the QTRAP using extracted exact mass ion current profiles for guanfacine and its stable-isotope-incorporated internal standard. The sample preparation employed liquid-liquid extraction with methyl t-butyl ether of 5 mm punched DBS card disks, followed by reversed-phase HPLC separation coupled with either MS/MS or high-resolution MS. Routine experiments were performed to establish the robustness of the DBS assay, including precision, accuracy, linearity, selectivity, sensitivity and long-term stability of up to 76 days. In addition, several factors that potentially affect quantitation were investigated, including blood volume for DBS spotting, punch size and punch location. A sensitive research assay with a LLOQ of 0.05 ng/ml was developed and subjected to several components of a method validation common to a regulated bioanalysis procedure employing DBS. This method development and partial validation study determined that spot volume, punch size or punch location do not affect assay accuracy and precision. The DBS approach was successfully applied to a clinical study (a Phase I, randomized, double-blind, placebo-controlled, crossover study to assess the effect of varying multiple oral doses of guanfacine on the pharmacokinetic, pharmacodynamic, safety, and tolerability profiles in healthy adult subjects). The pharmacokinetic profiles for 12 volunteers generated from the DBS assay and from a previously validated plasma assay were compared and were found to be comparable. DBS incurred samples collected from finger prick blood and

  2. Detection of Streptococcus mutans Genomic DNA in Human DNA Samples Extracted from Saliva and Blood

    PubMed Central

    Vieira, Alexandre R.; Deeley, Kathleen B.; Callahan, Nicholas F.; Noel, Jacqueline B.; Anjomshoaa, Ida; Carricato, Wendy M.; Schulhof, Louise P.; DeSensi, Rebecca S.; Gandhi, Pooja; Resick, Judith M.; Brandon, Carla A.; Rozhon, Christopher; Patir, Asli; Yildirim, Mine; Poletta, Fernando A.; Mereb, Juan C.; Letra, Ariadne; Menezes, Renato; Wendell, Steven; Lopez-Camelo, Jorge S.; Castilla, Eduardo E.; Orioli, Iêda M.; Seymen, Figen; Weyant, Robert J.; Crout, Richard; McNeil, Daniel W.; Modesto, Adriana; Marazita, Mary L.

    2011-01-01

    Caries is a multifactorial disease, and studies aiming to unravel the factors modulating its etiology must consider all known predisposing factors. One major factor is bacterial colonization, and Streptococcus mutans is the main microorganism associated with the initiation of the disease. In our studies, we have access to DNA samples extracted from human saliva and blood. In this report, we tested a real-time PCR assay developed to detect copies of genomic DNA from Streptococcus mutans in 1,424 DNA samples from humans. Our results suggest that we can determine the presence of genomic DNA copies of Streptococcus mutans in both DNA samples from caries-free and caries-affected individuals. However, we were not able to detect the presence of genomic DNA copies of Streptococcus mutans in any DNA samples extracted from peripheral blood, which suggests the assay may not be sensitive enough for this goal. Values of the threshold cycle of the real-time PCR reaction correlate with higher levels of caries experience in children, but this correlation could not be detected for adults. PMID:21731912

  3. Detection of Streptococcus mutans Genomic DNA in Human DNA Samples Extracted from Saliva and Blood.

    PubMed

    Vieira, Alexandre R; Deeley, Kathleen B; Callahan, Nicholas F; Noel, Jacqueline B; Anjomshoaa, Ida; Carricato, Wendy M; Schulhof, Louise P; Desensi, Rebecca S; Gandhi, Pooja; Resick, Judith M; Brandon, Carla A; Rozhon, Christopher; Patir, Asli; Yildirim, Mine; Poletta, Fernando A; Mereb, Juan C; Letra, Ariadne; Menezes, Renato; Wendell, Steven; Lopez-Camelo, Jorge S; Castilla, Eduardo E; Orioli, Iêda M; Seymen, Figen; Weyant, Robert J; Crout, Richard; McNeil, Daniel W; Modesto, Adriana; Marazita, Mary L

    2011-01-01

    Caries is a multifactorial disease, and studies aiming to unravel the factors modulating its etiology must consider all known predisposing factors. One major factor is bacterial colonization, and Streptococcus mutans is the main microorganism associated with the initiation of the disease. In our studies, we have access to DNA samples extracted from human saliva and blood. In this report, we tested a real-time PCR assay developed to detect copies of genomic DNA from Streptococcus mutans in 1,424 DNA samples from humans. Our results suggest that we can determine the presence of genomic DNA copies of Streptococcus mutans in both DNA samples from caries-free and caries-affected individuals. However, we were not able to detect the presence of genomic DNA copies of Streptococcus mutans in any DNA samples extracted from peripheral blood, which suggests the assay may not be sensitive enough for this goal. Values of the threshold cycle of the real-time PCR reaction correlate with higher levels of caries experience in children, but this correlation could not be detected for adults.

  4. Behavior of optical properties of coagulated blood sample at 633 nm wavelength

    NASA Astrophysics Data System (ADS)

    Morales Cruzado, Beatriz; Vázquez y Montiel, Sergio; Delgado Atencio, José Alberto

    2011-03-01

    Determination of tissue optical parameters is fundamental for application of light in either diagnostics or therapeutical procedures. However, in samples of biological tissue in vitro, the optical properties are modified by cellular death or cellular agglomeration that can not be avoided. This phenomena change the propagation of light within the biological sample. Optical properties of human blood tissue were investigated in vitro at 633 nm using an optical setup that includes a double integrating sphere system. We measure the diffuse transmittance and diffuse reflectance of the blood sample and compare these physical properties with those obtained by Monte Carlo Multi-Layered (MCML). The extraction of the optical parameters: absorption coefficient μa, scattering coefficient μs and anisotropic factor g from the measurements were carried out using a Genetic Algorithm, in which the search procedure is based in the evolution of a population due to selection of the best individual, evaluated by a function that compares the diffuse transmittance and diffuse reflectance of those individuals with the experimental ones. The algorithm converges rapidly to the best individual, extracting the optical parameters of the sample. We compare our results with those obtained by using other retrieve procedures. We found that the scattering coefficient and the anisotropic factor change dramatically due to the formation of clusters.

  5. Comparative analysis of RNA-Seq data from brain and blood samples of Parkinson's disease.

    PubMed

    Chatterjee, Paulami; Roy, Debjani

    2017-03-11

    Parkinson's disease (PD) is the second most common neurodegenerative disorders throughout the world. In order to search for PD biomarkers, we performed a system-level study of RNA-Seq data from PD brain and blood samples. Differentially expressed miRs of RNA-Seq data were subjected to generate the Co-expression networks. Three highly co-expressed clusters were identified based on their correlation coefficient values and fold change ratio. SM2miR drugs of the miRs contained in the three highly co-expressed clusters were identified, and drugs common among these clusters were selected. Co-expressed miRs not previously known to be associated with PD were identified from both the samples. Functional enrichment analyses of these miR targets were done, and the pathways common and unique to both the samples were identified. Thus, our study presents a comparative analysis of miRs, their associated pathways, and drugs from brain and blood samples of PD that may help in system level understanding of this disease. miRs identified from our study may serve as biomarkers for PD.

  6. Leukocyte count affects expression of reference genes in canine whole blood samples

    PubMed Central

    2011-01-01

    Background The dog is frequently used as a model for hematologic human diseases. In this study the suitability of nine potential reference genes for quantitative RT-PCR studies in canine whole blood was investigated. Findings The expression of these genes was measured in whole blood samples of 263 individual dogs, representing 73 different breeds and a group of 40 mixed breed dogs, categorized into healthy dogs and dogs with internal and hematological diseases, and dogs that underwent a surgical procedure. GeNorm analysis revealed that a combination of 5 to 6 of the most stably expressed genes constituted a stable normalizing factor. Evaluation of the expression revealed different ranking of reference genes in Normfinder and GeNorm. The disease category and the white blood cell count significantly affected reference gene expression. Conclusions The discrepancy between the ranking of reference genes in this study by Normfinder and Genorm can be explained by differences between the experimental groups such as "disease category" and "WBC count". This stresses the importance of assessing the expression stability of potential reference genes for gene experiments in canine whole blood anew for each specific experimental condition. PMID:21303565

  7. Assessment of malathion and its effects on leukocytes in human blood samples

    PubMed Central

    Sharma, Amit Kumar; Tiwari, Udita; Gaur, Mulayam Singh; Tiwari, Rajeev Kumar

    2016-01-01

    Abstract In the present paper, we report a reproducible, cost effective, fast response method for detection of malathion and its effects on leukocytes in different human blood groups. Spectroscopic methods (UV-Vis spectrometry) and Fourier transform infrared coupled with solid phase extraction were applied for analyzing malathion content in human blood plasma. The spiking levels of malathion in the range of 0.1-1.7 µg/mL were extracted from blood plasma samples using SPE. The present active functional groups (C = O; P-O-C; -OH; P = S) were also characterized. The recovery rate of malathion was 80%±4.5%. The calculated correlation coefficient was 0.9799, indicating the linearity of the results. The limit of detection (LOD) and limit of quantification (LOQ) were (0.1-1.7) µg/mL and (0.3-1.5) µg/mL, respectively. Malathion <1.0 µg/mL showed no significant change while higher levels of malathion exposure (1.5 µg/mL and 3.0 µg/mL) reduced the number of white blood cells. In conclusion, the spectroscopic results may be useful to understand the mechanism of other pesticides such as methyl parathion and parathion.

  8. Sensitivity of PCR Assays for Murine Gammaretroviruses and Mouse Contamination in Human Blood Samples

    PubMed Central

    Lee, Li Ling; Lin, Lin; Bell, David S.; Levine, Susan; Hanson, Maureen R.

    2012-01-01

    Gammaretroviruses related to murine leukemia virus (MLV) have variously been reported to be present or absent in blood from chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) patients and healthy controls. Using subjects from New York State, we have investigated by PCR methods whether MLV-related sequences can be identified in nucleic acids isolated from whole blood or from peripheral blood mononuclear cells (PBMCs) or following PBMC culture. We have also passaged the prostate cancer cell line LNCaP following incubation with plasma from patients and controls and assayed nucleic acids for viral sequences. We have used 15 sets of primers that can effectively amplify conserved regions of murine endogenous and exogenous retrovirus sequences. We demonstrate that our PCR assays for MLV-related gag sequences and for mouse DNA contamination are extremely sensitive. While we have identified MLV-like gag sequences following PCR on human DNA preparations, we are unable to conclude that these sequences originated in the blood samples. PMID:22629404

  9. Gingival crevicular blood: As a non-invasive screening tool for diabetes mellitus in dental clinics

    PubMed Central

    Shetty, Neema; Shankarapillai, Rajesh; Mathur, Lalit Kumar; Manohar, Balaji; Mathur, Aditi; Jain, Meetu

    2013-01-01

    Background: A high number of patients with periodontitis may have undiagnosed diabetes. Self-monitoring devices provide a simple method for rapid monitoring of the glucose level in the blood by utilizing a blood sample from the finger, but this method requires a needle puncture to obtain blood. It is possible that gingival crevicular blood (GCB) from routine periodontal probing may be a source of blood for glucose measurements. Aim: To establish whether GCB can be used as a non-invasive diagnostic aid in screening for diabetes mellitus during routine periodontal examination. Materials and Methods: The study involved 50 diabetics and 50 non-diabetics, with an age range of 26-66 years. Both diabetic and non-diabetic patients had moderate to severe gingivitis with at least one tooth in the maxillary anterior region showing bleeding upon probing. The Gingival Index and Oral Hygiene Index-Simplified were recorded. Blood oozing from the gingival sulcus/pocket following periodontal pocket probing was collected using a capillary tube and transferred to the test stick of a glucose self-monitoring device (Accu-Chek, Roche Diagnostic, Germany) in patients with comparable gingival and oral hygiene status. This value was compared with the peripheral fingerstick blood glucose (PFBG) value, which was obtained by pricking the finger tip at the same visit. Statistical analysis was performed using Pearson's correlation coefficient. Result: There was no statistically significant difference between the gingival crevicular blood glucose (GCBG) values and the PFBG values in both the diabetic (P = 0.129, NS) and the non-diabetic (P = 0.503, NS) groups. Karl Pearson's product–moment correlation coefficient was calculated, which showed a positive correlation between the two measurements in the diabetic (r = 0.943) as well as the non-diabetic (r = 0.926) groups. Conclusion: The results suggest that GCB can be used as a non-invasive diagnostic aid in screening for diabetes mellitus during

  10. Evaluation of a novel dried blood spot collection device (HemaSpot™) to test blood samples collected from dogs for antibodies to Leishmania infantum.

    PubMed

    Rosypal, Alexa C; Pick, Leanne D; Hernandez, Jaime O Esquivel; Lindsay, David S

    2014-09-15

    Collection of blood samples from veterinary and wildlife patients is often challenging because the samples have to be collected on farm or in the wild under various environmental conditions. This poses many technical problems associated with venipuncture materials, their safe use and disposal, transportation and processing of collected samples. Dried blood spot (DBS) sample collection techniques offer a simple and practical alternative to traditional blood collection methods to obtain blood samples from animals for parasite antibody evaluation. The DBS collection devices are compact, simple to use, and are particularly useful for large number of samples. Additionally, DBS samples take up less space and they are easier to transport than traditional venipuncture-collected blood samples. Visceral leishmaniasis (VL) is a potentially fatal parasitic disease of dogs and humans and it is frequently diagnosed by antibody tests. Immunochromatographic tests (ICT) for antibodies to Leishmania infantum are commercially available for dogs and they produce qualitative results in minutes. Measurement of canine antibodies to L. infantum with the ICT using traditional venipuncture has been validated previously, but the use of DBS samples has not been evaluated using this method. The purpose of the present study was to determine the ability of DBS samples to detect antibodies to L. infantum in dogs using a commercial ICT assay. One hundred plasma samples from dogs experimentally infected with the LIVT-1 strain of L. infantum were collected by venipuncture and frozen. Individual samples were thawed, and then 80 μl plasma (2 drops) was aliquotted onto the 8-spoked disk pad on individual DBS sample collection devices (HemaSpot™, Spot-On Sciences, Austin, TX), dried, and stored in the dark at room temperature. After one month and six months, respectively, 2 spokes of the 8 spokes of the disk pad of each DBS sample were removed and eluted in 200 μl PBS. The eluate was used to test

  11. Development of implantable autologous small-calibre vascular grafts from peripheral blood samples.

    PubMed

    Aper, T; Teebken, O E; Krüger, A; Heisterkamp, A; Hilfiker, A; Haverich, A

    2013-04-01

    At present the generation of a small-calibre (≤5 mm) vascular replacement for artificial bypasses remains a challenge for tissue engineering. The biocompatibility of bioartificial vessel replacements is of decisive significance for function and depends on the materials used. A completely autologous vessel substitute must exhibit high biocompatibility and functionality. For this purpose we developed and optimised a technique for the engineering of an autologous bypass material from a fibrin scaffold and vascular cells isolated from the same sample of peripheral blood in a porcine model. Fibrinogen, late outgrowth endothelial and smooth muscle cells were isolated from peripheral blood samples (n=14, 100 mL each). Fibroblasts were isolated from porcine aortic adventitial tissue (n=4). Tubular seeded fibrin segments were obtained using an injection moulding technique with the simultaneous incorporation of the in vitro expanded cells into the fibrin matrix. The segments were cultivated under dynamic conditions with pulsatile perfusion in a bioreactor. Morphological and functional characterization was done. Artificial vascular segments with a length of 150 mm were reproducibly obtained with a hierarchical arrangement of incorporated cells similar to the structure of the vascular wall. By additional seeding of fibroblasts, suturable segments with biomechanical properties suitable for implantation into the arterial system were obtained. Implantable bioartificial vascular grafts can be generated from blood. After cultivation under dynamic conditions the vascular segments possess a structure similar to that of the vascular wall and exhibit biomechanical properties sufficient for implantation as arterial substitutes. Georg Thieme Verlag KG Stuttgart · New York.

  12. Determination of DDT and related compounds in blood samples from agricultural workers.

    PubMed

    Guardino, X; Serra, C; Obiols, J; Rosell, M G; Berenguer, M J; López, F; Brosa, J

    1996-01-05

    An analytical method combining a solid-phase (C18) clean-up and GC-electron-capture detection using a capillary column, was implemented to determine p,p'-DDT and its metabolites (p,p'-DDD and p,p'-DDE), as well as other organochlorine pesticides in whole blood samples from 30 farmers and 24 non-occupationally exposed workers. The average concentrations for the quantified pesticides, p,p'-DDT, p,p'-DDD and p,p'-DDE, were 0.9, 1.5 and 8.0 micrograms/l whole blood for exposed workers and 0.3, 0.5 and 3.3 micrograms/l for unexposed workers, respectively. GC-MS was used to confirm the identity of the pesticides found. Solid-phase extraction and the protocol used give a cleaner analytical matrix, not only improving sensitivity and resolution, but also allowing analyses with smaller blood samples as compared to other methods.

  13. Poppy seed consumption and toxicological analysis of blood and urine samples.

    PubMed

    Moeller, Manfred R; Hammer, Karin; Engel, Oliver

    2004-07-16

    Poppy seeds contain morphine in different amounts. Reported concentrations are up to 294 mg morphine/kg poppy seeds. Since penalties based on Street Traffic Law (parapgraph 24a StVG) in Germany (administrative offence) require definitive proof of morphine in blood samples, and the "Grenzwertkommission" in consultation with the Ministry of Transportation recommended a threshold of free morphine of 10 ng/mL, the question arose whether the consumption of poppy seeds can lead to a blood concentrations equal or higher than 10 ng/mL of free morphine. Therefore, five volunteers ate poppy seed products (50 mg morphine/kg poppy seeds). In urine, all on-site tests were enzyme immunologically positive for opiates and were positive to morphine by GC/MS. All the blood samples were negative to morphine by EIA and to free morphine by GC/MS. However, after hydrolysis, morphine was detected by GC/MS in all cases. Accordingly, in Germany, penalties based on parapgraph 24a StVG are not likely to cause road users any concerns should they have consumed poppy seeds. Driver Licensing Authorities, however, should be advised of this problem to avoid unjustified legal measures.

  14. Application of Atomic Dielectric Resonance Spectroscopy for the screening of blood samples from patients with clinical variant and sporadic CJD

    PubMed Central

    Fagge, Timothy J; Barclay, G Robin; Stove, G Colin; Stove, Gordon; Robinson, Michael J; Head, Mark W; Ironside, James W; Turner, Marc L

    2007-01-01

    Background Sub-clinical variant Creutzfeldt-Jakob disease (vCJD) infection and reports of vCJD transmission through blood transfusion emphasise the need for blood screening assays to ensure the safety of blood and transplanted tissues. Most assays aim to detect abnormal prion protein (PrPSc), although achieving required sensitivity is a challenge. Methods We have used innovative Atomic Dielectric Resonance Spectroscopy (ADRS), which determines dielectric properties of materials which are established by reflectivity and penetration of radio/micro waves, to analyse blood samples from patients and controls to identify characteristic ADR signatures unique to blood from vCJD and to sCJD patients. Initial sets of blood samples from vCJD, sCJD, non-CJD neurological diseases and normal healthy adults (blood donors) were screened as training samples to determine group-specific ADR characteristics, and provided a basis for classification of blinded sets of samples. Results Blood sample groups from vCJD, sCJD, non-CJD neurological diseases and normal healthy adults (blood donors) screened by ADRS were classified with 100% specificity and sensitivity, discriminating these by a co-variance expert analysis system. Conclusion ADRS appears capable of recognising and discriminating serum samples from vCJD, sCJD, non-CJD neurological diseases, and normal healthy adults, and might be developed to provide a system for primary screening or confirmatory assay complementary to other screening systems. PMID:17760958

  15. Blood Sampling Seasonality as an Important Preanalytical Factor for Assessment of Vitamin D Status

    PubMed Central

    Bonelli, Patrizia; Buonocore, Ruggero; Aloe, Rosalia

    2016-01-01

    Summary Background The measurement of vitamin D is now commonplace for preventing osteoporosis and restoring an appropriate concentration that would be effective to counteract the occurrence of other human disorders. The aim of this study was to establish whether blood sampling seasonality may influence total vitamin D concentration in a general population of Italian unselected outpatients. Methods We performed a retrospective search in the laboratory information system of the University Hospital of Parma (Italy, temperate climate), to identify the values of total serum vitamin D (25-hydroxyvitamin D) measured in outpatients aged 18 years and older, who were referred for routine health check-up during the entire year 2014. Results The study population consisted in 11,150 outpatients (median age 62 years; 8592 women and 2558 men). The concentration of vitamin D was consistently lower in samples collected in Winter than in the other three seasons. The frequency of subjects with vitamin D deficiency was approximately double in samples drawn in Winter and Spring than in Summer and Autumn. In the multivariate analysis, the concentration of total vitamin D was found to be independently associated with sex and season of blood testing, but not with the age of the patients. Conclusions According to these findings, blood sampling seasonality should be regarded as an important preanalytical factor in vitamin D assessment. It is also reasonable to suggest that the amount of total vitamin D synthesized during the summer should be high enough to maintain the levels > 50 nmol/L throughout the remaining part of the year. PMID:28356869

  16. Improved age determination of blood and teeth samples using a selected set of DNA methylation markers

    PubMed Central

    Kamalandua, Aubeline

    2015-01-01

    Age estimation from DNA methylation markers has seen an exponential growth of interest, not in the least from forensic scientists. The current published assays, however, can still be improved by lowering the number of markers in the assay and by providing more accurate models to predict chronological age. From the published literature we selected 4 age-associated genes (ASPA, PDE4C, ELOVL2, and EDARADD) and determined CpG methylation levels from 206 blood samples of both deceased and living individuals (age range: 0–91 years). This data was subsequently used to compare prediction accuracy with both linear and non-linear regression models. A quadratic regression model in which the methylation levels of ELOVL2 were squared showed the highest accuracy with a Mean Absolute Deviation (MAD) between chronological age and predicted age of 3.75 years and an adjusted R2 of 0.95. No difference in accuracy was observed for samples obtained either from living and deceased individuals or between the 2 genders. In addition, 29 teeth from different individuals (age range: 19–70 years) were analyzed using the same set of markers resulting in a MAD of 4.86 years and an adjusted R2 of 0.74. Cross validation of the results obtained from blood samples demonstrated the robustness and reproducibility of the assay. In conclusion, the set of 4 CpG DNA methylation markers is capable of producing highly accurate age predictions for blood samples from deceased and living individuals PMID:26280308

  17. Effect of Ibuprofen dose on platelet aggregation and coagulation in blood samples from pigs.

    PubMed

    Martini, Wenjun Z; Deguzman, Rodolfo; Rodriguez, Cassandra M; Guerra, Jessica; Martini, Angela K; Pusateri, Anthony E; Dubick, Michael A

    2015-03-01

    Ibuprofen is commonly used by Soldiers in the deployed environment. This study investigated its dose-effects on in vitro coagulation. Blood samples were collected from 4 normal healthy pigs and were processed to make platelet-adjusted (100×10(3)/μL) blood samples. Ibuprofen was added to the samples at doses of 0 μg/mL (control), recommended oral dose (163 μg/mL, 1×), 2×, 4×, 8×, 10×, 12×, 16×, and 20×. Arachidonic acid or collagen-stimulated platelet aggregation was assessed at 15 minutes after the addition of ibuprofen. Coagulation was assessed with measurements of prothrombin time (PT) and activated partial thromboplastin time (aPTT), and thrombelastography by Rotem. A robust inhibition of ibuprofen on arachidonic acid-induced platelet aggregation was observed at all doses tested. Collagen-stimulated platelet aggregation was inhibited to 71%±5% and 10%±5% of the control values at ibuprofen doses of 4× and 20×, respectively (both p<0.05). No changes were observed in PT at any dose, but aPTT was prolonged at dose of 16× and 20×. Rotem measurements of coagulation time, clot formation time, maximum clot firmness, and A10 were compromised at dose 16× and 20× (all p<0.05). Ibuprofen inhibited platelet aggregation at recommended doses, but did not compromise aPTT or coagulation profile until at 16 times the recommended doses and higher. Further effort is needed to clarify whether there are different dose-responses between human and pig blood samples in trauma situations. Reprint & Copyright © 2015 Association of Military Surgeons of the U.S.

  18. Blood

    MedlinePlus

    ... The liquid part, called plasma, is made of water, salts, and protein. Over half of your blood is plasma. The solid part of your blood contains red blood cells, white blood cells, and platelets. Red ...

  19. First Molecular Identification of Dirofilaria repens in a Dog Blood Sample from Guanajuato, Mexico.

    PubMed

    Ramos-Lopez, Sandra; León-Galván, Ma Fabiola; Salas-Alatorre, Mariana; Lechuga-Arana, Alma A; Valencia-Posadas, Mauricio; Gutiérrez-Chávez, Abner J

    2016-11-01

    Dirofilaria repens is the causative zoonotic agent of subcutaneous dirofilariosis. Although the American continent is considered currently free of D. repens infections, in the past few years there have been some reports suggesting the presence of this agent in the Americas. In Guanajuato, Mexico, there is a report of the presence of D. repens. To determine whether this parasite is really present in Guanajuato, a total of 177 EDTA blood samples from dogs were tested using the modified Knott's technique. Using this technique, we found 15 positive samples, which were then analyzed using the PCR technique, with a panfilarial set of primers and a specific primer pair for D. repens. In one sample, the expected band was present and the corresponding amplicon, sequenced, displayed a 100% identity to D. repens, confirming the presence of this exotic and zoonotic filarial species in the municipality of Silao, Guanajuato, Mexico.

  20. Direct identification of bacteria in blood culture samples using an electronic nose.

    PubMed

    Trincavelli, Marco; Coradeschi, Silvia; Loutfi, Amy; Söderquist, Bo; Thunberg, Per

    2010-12-01

    In this paper, we introduce a method for identification of bacteria in human blood culture samples using an electronic nose. The method uses features, which capture the static (steady state) and dynamic (transient) properties of the signal from the gas sensor array and proposes a means to ensemble results from consecutive samples. The underlying mechanism for ensembling is based on an estimation of posterior probability, which is extracted from a support vector machine classifier. A large dataset representing ten different bacteria cultures has been used to validate the presented methods. The results detail the performance of the proposed algorithm and show that through ensembling decisions on consecutive samples, significant reliability in classification accuracy can be achieved.

  1. Trace element levels in whole blood samples from residents of the city Badajoz, Spain.

    PubMed

    Moreno, M A; Marin, C; Vinagre, F; Ostapczuk, P

    1999-05-19

    Copper, lead, cadmium, and zinc were determined by anodic stripping voltammetry after sample digestion and potentiometric stripping analysis was used for Pb and Cd determination in original samples. Selenium was determined by cathodic stripping voltammetry or hydride generation AAS. Element levels found in the whole blood sample in a group of 82 people are for Cd: 0.98 +/- 0.94 ng/ml; for Pb: 46.7 +/- 28.6 ng/ml; for Cu: 1.07 +/- 0.12 micrograms/ml; for Zn: 6.95 +/- 1.08 micrograms/ml, and for Se: 116 +/- 25 ng/ml. Analytical data have been correlated to age, sex, smokers or non-smokers, drinking and food habits.

  2. Effects of repeated blood samplings on locomotor activity, evasion and wheel-running activity in mice.

    PubMed

    Pfeil, R

    1988-01-01

    The effects of serial blood sampling on nocturnal locomotor activity, evasion, wheel-running activity and body mass were studied in male NMRI mice aged 7-8 weeks. The erythrocyte count, haematocrit and haemoglobin concentration at the beginning and end of the study showed no difference in group 1 (two samples per week, 0.08 ml each) while there was a significant decrease in the group 2 values (three samples per week, 0.08 ml each). The total amount of nocturnal locomotor activity decreased in the animals bled repeatedly while the periods with locomotor activity increased. These alterations appeared particularly after bleeding. In the test-group animals evasion showed a decrease compared with the untreated control animals, but there was no evidence of a relation to the timing of the bleedings.

  3. Less invasive blood sampling in the animal laboratory: clinical chemistry and haematology of blood obtained by the Triatominae bug Dipetalogaster maximus.

    PubMed

    Markvardsen, S N; Kjelgaard-Hansen, M; Ritz, C; Sørensen, D B

    2012-04-01

    Dipetalogaster maximus (Dipmax), a blood-sucking bug belonging to the family Reduviidae, has been used to obtain blood samples, for example for clinical chemistry and haematology, in a variety of zoo animals and wildlife. Using this bug allows stress-free blood sampling as the bug is able to draw blood without the mammal noticing the bug. In laboratory animal science, the need for blood samples from unstressed animals may arise, especially in animal behaviour research. The use of Dipmax bugs may prove a valuable tool for this purpose. To validate the method, we compared an array of standard blood parameters sampled from New Zealand White rabbits, sampled either by the use of bugs or by the conventional method; puncture of vena auricularis caudalis. The overall hypothesis was that there was no significant difference in clinical chemistry and haematological parameters between the bug method and the conventional method. A total of 17 clinical parameters as well as 12 haematological parameters were measured and compared in New Zealand White rabbits. The results showed that for 13 of these 29 analysed parameters, the bug method and the conventional method did not give significantly different results, and the obtained results were thus directly comparable. For the remaining parameters the obtained results were significantly different. However, all parameters were measurable in the bug samples. The influences of the bug metabolism on these parameters are discussed.

  4. Blood pressure interacts with APOE ε4 to predict memory performance in a midlife sample.

    PubMed

    Oberlin, Lauren E; Manuck, Stephen B; Gianaros, Peter J; Ferrell, Robert E; Muldoon, Matthew F; Jennings, J Richard; Flory, Janine D; Erickson, Kirk I

    2015-09-01

    Elevated blood pressure and the Apolipoprotein ε4 allele (APOE ε4) are independent risk factors for Alzheimer's disease. We sought to determine whether the combined presence of the APOE ε4 allele and elevated blood pressure is associated with lower cognitive performance in cognitively healthy middle-aged adults. A total of 975 participants aged 30-54 (mean age = 44.47) were genotyped for APOE. Cardiometabolic risk factors including blood pressure, lipids, and glucose were assessed and cognitive function was measured using the Trail Making Test and the Visual Reproduction and Logical Memory subtests from the Wechsler Memory Scale. Multivariable regression analysis showed that the association between APOE ε4 and episodic memory performance varied as a function of systolic blood pressure (SBP), such that elevated SBP was predictive of poorer episodic memory performance only in APOE ε4 carriers (β = -.092; t = -2.614; p = .009). Notably, this association was apparent at prehypertensive levels (≥130 mmHg), even after adjusting for physical activity, depression, smoking, and other cardiometabolic risk factors. The joint presence of APOE ε4 and elevated SBP, even at prehypertensive levels, is associated with lower cognitive performance in healthy, middle-aged adults. Results of this study suggest that the combination of APOE ε4 and elevated SBP may synergistically compromise memory function well before the appearance of clinically significant impairments. Interventions targeting blood pressure control in APOE ε4 carriers during midlife should be studied as a possible means to reduce the risk of cognitive decline in genetically susceptible samples. (c) 2015 APA, all rights reserved).

  5. Micro-method for the determination of glutathione in human blood.

    PubMed

    Giustarini, Daniela; Fanti, Paolo; Matteucci, Elena; Rossi, Ranieri

    2014-08-01

    A new procedure is described for the visible-range spectrophotometric analysis of glutathione (GSH) in microvolumes of blood (as low as 0.5 μL) collected by fingerstick. Samples are diluted 1 to 300 (v/v) in a stabilizing solution, followed by determination of haemoglobin concentration and by acid deproteination. GSH is then measured in the clear supernatant by colorimetry using DTNB, i.e., 5,5'-dithio-bis(2-nitrobenzoic acid), in aqueous solution at pH 7.8. The DTNB reagent is prepared and kept at pH 6.2 until just prior its addition, thus avoiding spontaneous decomposition of the reagent. The assay is rapid, easy to adapt to large-scale studies and it avoids artefactual oxidation of GSH, a common methodological shortcoming. The method is precise with 1.7 to 3.4% intra-day relative standard deviation (RSD) and 2.2 to 4.2% inter-day RSD, and accurate with -1.4% to 2.3% intra-day relative error (RE) and -2.8% to 1.6% inter-day RE. GSH is recovered by 97.5 to 100% at all tested concentrations. The new colorimetric micro-method was validated by a reliable previously reported HPLC method. The procedure is suitable for minimally invasive investigation of oxidative stress in peripheral blood. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Luminol chemiluminescence biosensor for glycated hemoglobin (HbA1c) in human blood samples.

    PubMed

    Ahn, Kwang-Soo; Lee, JungHoon; Park, Jong-Myeon; Choi, Han Nim; Lee, Won-Yong

    2016-01-15

    Luminol chemiluminescence (CL) biosensor based on boronic acid modified gold substrate has been developed for the determination of glycated hemoglobin (HbA1c) in human blood samples. In order to selectively capture HbA1c in sample, carboxy-EG6-undecanethiol was self-assembled on a gold thin-film substrate, followed by covalent coupling of 3-aminophenyl boronic acid (3-APBA). The captured HbA1c containing four iron heme groups plays as a catalyst for luminol CL reaction in the presence of hydrogen peroxide, and thus the luminol CL response is linearly proportional to the amount of HbA1c captured on the biosensor surface. The present biosensor showed linear dynamic range of HbA1c from 2.5% to 17.0%, which well covers the clinically important concentration range. In addition, the present biosensor exhibited negligible response to interfering species such as hemoglobin, fructose, and sorbitol. The present HbA1c biosensor was applied to the determination of HbA1c in human blood samples and the results were well agreed with that obtained with a conventional method.

  7. Chromatographic resolution, characterisation and quantification of VX enantiomers in hemolysed swine blood samples.

    PubMed

    Reiter, Georg; Mikler, John; Hill, Ira; Weatherby, Kendal; Thiermann, Horst; Worek, Franz

    2008-09-15

    The present study was initiated to develop a sensitive and highly selective method for the analysis of the enantiomers of the nerve agent VX (O-ethyl S-[2(diisopropylamino)ethyl] methylphosphonothioate) in blood samples for toxicokinetic and therapeutic research. To achieve this goal, analytical and semi-preparative enantioseparation of VX were carried out with gas and liquid chromatography. The GC chiral stationary phase was HYDRODEX-beta-TBDAc (beta cyclodextrin), on which VX was baseline-resolved. On the chiral HPLC phase CHIRALCEL OD-H the enantiomers of VX were isolated with enantiomeric excess >99.99%. They were characterised by specific optical rotation (+/-25.8 deg ml dm(-1)g(-1) at 20 degrees C and 589 nm) and by determination of cholinesterase inhibition rate constants. For the quantitative chiral detection of VX the enantioresolution was realized on the HPLC chiral phase CHIRAL AGP. A specific procedure was developed to isolate VX from swine blood samples thereby stabilising its enantiomers. The limit of detection was 200 fg per enantiomer on column. The absolute recovery of the overall sample preparation procedure was 75%. After an intravenous and percutaneous administration of a supralethal dose of VX in anesthetised swine (+)-VX and (-)-VX could be quantified up to 720 min.

  8. Capillary blood sampling: national recommendations on behalf of the Croatian Society of Medical Biochemistry and Laboratory Medicine.

    PubMed

    Krleza, Jasna Lenicek; Dorotic, Adrijana; Grzunov, Ana; Maradin, Miljenka

    2015-01-01

    Capillary blood sampling is a medical procedure aimed at assisting in patient diagnosis, management and treatment, and is increasingly used worldwide, in part because of the increasing availability of point-of-care testing. It is also frequently used to obtain small blood volumes for laboratory testing because it minimizes pain. The capillary blood sampling procedure can influence the quality of the sample as well as the accuracy of test results, highlighting the need for immediate, widespread standardization. A recent nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia has shown that capillary sampling procedures are not standardized and that only a small proportion of Croatian laboratories comply with guidelines from the Clinical Laboratory Standards Institute (CLSI) or the World Health Organization (WHO). The aim of this document is to provide recommendations for capillary blood sampling. This document has been produced by the Working Group for Capillary Blood Sampling within the Croatian Society of Medical Biochemistry and Laboratory Medicine. Our recommendations are based on existing available standards and recommendations (WHO Best Practices in Phlebotomy, CLSI GP42-A6 and CLSI C46-A2), which have been modified based on local logistical, cultural, legal and regulatory requirements. We hope that these recommendations will be a useful contribution to the standardization of capillary blood sampling in Croatia.

  9. Capillary blood sampling: national recommendations on behalf of the Croatian Society of Medical Biochemistry and Laboratory Medicine

    PubMed Central

    Krleza, Jasna Lenicek; Dorotic, Adrijana; Grzunov, Ana; Maradin, Miljenka

    2015-01-01

    Capillary blood sampling is a medical procedure aimed at assisting in patient diagnosis, management and treatment, and is increasingly used worldwide, in part because of the increasing availability of point-of-care testing. It is also frequently used to obtain small blood volumes for laboratory testing because it minimizes pain. The capillary blood sampling procedure can influence the quality of the sample as well as the accuracy of test results, highlighting the need for immediate, widespread standardization. A recent nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia has shown that capillary sampling procedures are not standardized and that only a small proportion of Croatian laboratories comply with guidelines from the Clinical Laboratory Standards Institute (CLSI) or the World Health Organization (WHO). The aim of this document is to provide recommendations for capillary blood sampling. This document has been produced by the Working Group for Capillary Blood Sampling within the Croatian Society of Medical Biochemistry and Laboratory Medicine. Our recommendations are based on existing available standards and recommendations (WHO Best Practices in Phlebotomy, CLSI GP42-A6 and CLSI C46-A2), which have been modified based on local logistical, cultural, legal and regulatory requirements. We hope that these recommendations will be a useful contribution to the standardization of capillary blood sampling in Croatia. PMID:26524965

  10. Evaluation of the validity of a rapid method for measuring high and low haemoglobin levels in whole blood donors.

    PubMed

    Shahshahani, Hayedeh J; Meraat, Nahid; Mansouri, Fatemeh

    2013-07-01

    Haemoglobin screening methods need to be highly sensitive to detect both low and high haemoglobin levels and avoid unnecessary rejection of potential blood donors. The aim of this study was to evaluate the accuracy of measurements by HemoCue in blood donors. Three hundred and fourteen randomly selected, prospective blood donors were studied. Single fingerstick blood samples were obtained to determine the donors' haemoglobin levels by HemoCue, while venous blood samples were drawn for measurement of the haemoglobin level by both HemoCue and an automated haematology analyser as the reference method. The sensitivity, specificity, predictive values and correlation between the reference method and HemoCue were assessed. Cases with a haemoglobin concentration in the range of 12.5-17.9 g/dL were accepted for blood donation. Analysis of paired results showed that haemoglobin levels measured by HemoCue were higher than those measured by the reference method. There was a significant correlation between the reference method and HemoCue for haemoglobin levels less than 12.5 g/dL. The correlation was less strong for increasing haemoglobin levels. Linear correlation was poor for haemoglobin levels over 18 g/dL. Thirteen percent of donors, who had haemoglobin levels close to the upper limit, were unnecessarily rejected. HemoCue is suitable for screening for anaemia in blood donors. Most donors at Yazd are males and a significant percentage of them have haemoglobin values close to the upper limit for acceptance as a blood donor; since these subjects could be unnecessarily rejected on the basis of HemoCue results and testing with this method is expensive, it is recommended that qualitative methods are used for primary screening and accurate quantitative methods used in clinically suspicious cases or when qualitative methods fail.

  11. The effect of different blood sampling sites and analyses on the relationship between exercise intensity and 4.0 mmol.l-1 blood lactate concentration.

    PubMed

    Foxdal, P; Sjödin, A; Ostman, B; Sjödin, B

    1991-01-01

    The aim of the study was to examine whether the difference in lactate concentration in different blood fractions is of practical importance when using blood lactate as a test variable of aerobic endurance capacity. Ten male firefighters performed submaximally graded exercise on a cycle ergometer for 20-25 min. Venous and capillary blood samples were taken every 5 min for determination of haematocrit and lactate concentrations in plasma, venous and capillary blood. At the same time, expired air was collected in Douglas bags for determination of the oxygen consumption. A lactate concentration of 4.0 mmol.l-1 was used as the reference value to compare the oxygen consumption and exercise intensity when different types of blood specimen and sampling sites were used for lactate analysis. At this concentration the exercise intensity was 17% lower (P less than 0.01) when plasma lactate was compared to venous blood lactate, and 12% lower (P less than 0.05) when capillary blood lactate was used. Similar discrepancies were seen in oxygen consumption. The results illustrated the importance of standardizing sampling and handling of blood specimens for lactate determination to enable direct comparisons to be made among results obtained in different studies.

  12. Acetaminophen and meloxicam inhibit platelet aggregation and coagulation in blood samples from humans.

    PubMed

    Martini, Angela K; Rodriguez, Cassandra M; Cap, Andrew P; Martini, Wenjun Z; Dubick, Michael A

    2014-12-01

    Acetaminophen (Ace) and meloxicam (Mel) are the two types of analgesic and antipyretic medications. This study investigated the dose responses of acetaminophen and meloxicam on platelet aggregation and coagulation function in human blood samples. Blood samples were collected from six healthy humans and processed to make platelet-adjusted (100 × 10 cells/μl) blood samples. Acetaminophen (Tylenol, Q-PAP, 100 mg/ml) was added at the doses of 0 μg/ml (control), 214 μg/ml (the standard dose, 1 ×), 4 ×, 8 ×, 10 ×, 12 ×, 16 ×, and 20 ×. Similarly, meloxicam (Metacam, 5 mg/ml) was added at doses of 0 μg/ml (control), 2.85 μg/ml (the standard dose, 1 ×), 4 ×, 8 ×, 10 ×, 12 ×, 16 ×, and 20 ×. Fifteen minutes after the addition of acetaminophen and/or meloxicam, platelet aggregation was stimulated with collagen (2 μg/ml) or arachidonic acid (0.5 mmol/l) and assessed using a Chrono-Log 700 aggregometer. Coagulation function was assessed by prothrombin time (PT), activated partial thromboplastin time (aPTT), and using Rotem thrombelastogram. A robust inhibition by acetaminophen and/or meloxicam was observed in arachidonic acid-stimulated platelet aggregation starting at 1 × dose. Collagen-stimulated platelet aggregation was inhibited by ACE starting at 1 × (78 ± 10% of control), and by meloxicam starting at 4 × (72 ± 5% of control, both P < 0.05). The inhibitions by acetaminophen and meloxicam combined were similar to those by acetaminophen or meloxicam. aPTT was prolonged by meloxicam starting at 4 ×. No changes were observed in PT or any of Rotem measurements by acetaminophen and/or meloxicam. Acetaminophen and meloxicam compromised platelet aggregation and aPTT. Further effort is warranted to characterize the effects of acetaminophen and meloxicam on bleeding in vivo.

  13. Selective Venous Blood Sampling for Hyperparathyroidism with unclear Localization of the Parathyroid Gland.

    PubMed

    Hader, C; Uder, M; Loose, R W R; Linnemann, U; Bertsch, T; Adamus, R

    2016-12-01

    Purpose: Evaluation of the benefit of selective venous blood sampling (SVS) for the preoperative identification of parathyroid adenomas with unclear localization in non-invasive diagnostics. Materials and Methods: In a retrospective study, all patients (n = 23) with primary (n = 21) or tertiary (n = 2) hyperparathyroidism were evaluated from 2005 to 2016 at the Hospital Nuremberg-North. These patients all received one (n = 20) or more (n = 3) SVS. 15 patients had one or more previous unsuccessful surgeries (group A), 8 patients received the SVS primarily before the first surgery (group B). Results of SVS were compared with the results of surgery, non-invasive diagnostic procedures and clinical follow up. Results: In 24 out of 26 SVS a significant PTH peak was found. 19 patients underwent surgery after SVS. In 16 of these cases (84 %) the SVS peak was concordant with the intraoperative localization. Thus, SVS of all operated patients had a sensitivity of 94 %. Considering only patients with prior HPT surgery the sensitivity was 89 %. In none of the 26 examinations complications occurred. Conclusion: Our results demonstrate that selective venous blood sampling SVS in cases with unclear imaging of parathyroid adenomas is an effective and low-risk invasive diagnostic method to localize parathyroid adenomas and helps to improve surgical therapy. Key points: • low risk invasive diagnostic procedure to localize parathyroid adenomas• additional step if non-invasive diagnostics are negative or inconclusive• high sensitivity in the detection of parathyroid adenomas Citation Format: • Hader C, Uder M, Loose RWR et al. Selective Venous Blood Sampling for Hyperparathyroidism with unclear Localization of the Parathyroid Gland. Fortschr Röntgenstr 2016; 188: 1144 - 1150. © Georg Thieme Verlag KG Stuttgart · New York.

  14. Does Pneumatic Tube System Transport Contribute to Hemolysis in ED Blood Samples?

    PubMed Central

    Phelan, Michael P.; Reineks, Edmunds Z.; Hustey, Fredric M.; Berriochoa, Jacob P.; Podolsky, Seth R.; Meldon, Stephen; Schold, Jesse D.; Chamberlin, Janelle; Procop, Gary W.

    2016-01-01

    Introduction Our goal was to determine if the hemolysis among blood samples obtained in an emergency department and then sent to the laboratory in a pneumatic tube system was different from those in samples that were hand-carried. Methods The hemolysis index is measured on all samples submitted for potassium analysis. We queried our hospital laboratory database system (SunQuest®) for potassium results for specimens obtained between January 2014 and July 2014. From facility maintenance records, we identified periods of system downtime, during which specimens were hand-carried to the laboratory. Results During the study period, 15,851 blood specimens were transported via our pneumatic tube system and 92 samples were hand delivered. The proportions of hemolyzed specimens in the two groups were not significantly different (13.6% vs. 13.1% [p=0.90]). Results were consistent when the criterion was limited to gross (3.3% vs 3.3% [p=0.99]) or mild (10.3% vs 9.8% [p=0.88]) hemolysis. The hemolysis rate showed minimal variation during the study period (12.6%–14.6%). Conclusion We found no statistical difference in the percentages of hemolyzed specimens transported by a pneumatic tube system or hand delivered to the laboratory. Certain features of pneumatic tube systems might contribute to hemolysis (e.g., speed, distance, packing material). Since each system is unique in design, we encourage medical facilities to consider whether their method of transport might contribute to hemolysis in samples obtained in the emergency department. PMID:27625719

  15. Does Pneumatic Tube System Transport Contribute to Hemolysis in ED Blood Samples?

    PubMed

    Phelan, Michael P; Reineks, Edmunds Z; Hustey, Fredric M; Berriochoa, Jacob P; Podolsky, Seth R; Meldon, Stephen; Schold, Jesse D; Chamberlin, Janelle; Procop, Gary W

    2016-09-01

    Our goal was to determine if the hemolysis among blood samples obtained in an emergency department and then sent to the laboratory in a pneumatic tube system was different from those in samples that were hand-carried. The hemolysis index is measured on all samples submitted for potassium analysis. We queried our hospital laboratory database system (SunQuest(®)) for potassium results for specimens obtained between January 2014 and July 2014. From facility maintenance records, we identified periods of system downtime, during which specimens were hand-carried to the laboratory. During the study period, 15,851 blood specimens were transported via our pneumatic tube system and 92 samples were hand delivered. The proportions of hemolyzed specimens in the two groups were not significantly different (13.6% vs. 13.1% [p=0.90]). Results were consistent when the criterion was limited to gross (3.3% vs 3.3% [p=0.99]) or mild (10.3% vs 9.8% [p=0.88]) hemolysis. The hemolysis rate showed minimal variation during the study period (12.6%-14.6%). We found no statistical difference in the percentages of hemolyzed specimens transported by a pneumatic tube system or hand delivered to the laboratory. Certain features of pneumatic tube systems might contribute to hemolysis (e.g., speed, distance, packing material). Since each system is unique in design, we encourage medical facilities to consider whether their method of transport might contribute to hemolysis in samples obtained in the emergency department.

  16. Filter-paper blood samples for ELISA detection of Brucella antibodies in caribou.

    PubMed

    Curry, Patricia S; Elkin, Brett T; Campbell, Mitch; Nielsen, Klaus; Hutchins, Wendy; Ribble, Carl; Kutz, Susan J

    2011-01-01

    We evaluated blood collected on Nobuto filter-paper (FP) strips for use in detecting Brucella spp. antibodies in caribou. Whole blood (for serum) and blood-saturated FP strips were obtained from 185 killed arctic caribou (Rangifer tarandus groenlandicus). Sample pairs (serum and FP eluates) were simultaneously tested in duplicate using competitive enzyme-linked immunosorbent assay (c-ELISA) and indirect ELISA (i-ELISA) for Brucella spp. Prior work based on isolation of Brucella spp. revealed sensitivity (SE) and specificity (SP) of 100% and 99%, respectively, for both these serum assays in caribou. Infection status of the animals in the current study was unknown but recent sampling had revealed clinical brucellosis and >40% Brucella antibody prevalence in the herd. To assess the performance of FP relative to serum in these assays, serum was used as the putative gold standard. On both assays, the findings for duplicate runs (A and B) were similar. For c-ELISA run A, the FP Brucella prevalence (47%) was lower than serum prevalence (52%), with SE 89% (95% confidence interval [CI]: 82-95%) and SP 99% (97-100%). For i-ELISA run A, serum and FP Brucella prevalence rates were identical (43%), and the SE and SP of FP testing were 100% and 99% (97-100%), respectively. The findings suggest better FP test performance with i-ELISA than with c-ELISA; however, i-ELISA does not distinguish cross-reacting antibodies induced by Brucella vaccination or exposure to certain other Gram-negative pathogens. Results for duplicate FP eluates (prepared using separate FP strips from each animal) were strongly correlated for both protocols (r=0.996 and 0.999 for c-ELISA and i-ELISA, respectively), indicating minimal variability among FPs from any individual caribou. Dried caribou FP blood samples stored for 2 mo at room temperature are comparable with serum for use in Brucella spp. c-ELISA and i-ELISA. Hunter-based FP sampling can facilitate detection of disease exposure in remote regions and

  17. Validation of a non-invasive blood-sampling technique for doubly-labelled water experiments.

    PubMed

    Voigt, Christian C; Helversen, Otto Von; Michener, Robert H; Kunz, Thomas H

    2003-04-01

    Two techniques for bleeding small mammals have been used in doubly-labeled water (DLW) studies, including vena puncture and the use of starved nymphal stages of hematophagous reduviid bugs (Reduviidae, Hemiptera). In this study, we tested the validity of using reduviid bugs in doubly-labeled water experiments. We found that the isotope enrichment in initial blood samples collected with bugs was significantly lower compared to isotope enrichment in blood samples obtained using vena puncture. We therefore used the desiccation method for estimating total body water (TBW) in DLW experiments because TBW calculated using the isotope dilution method was overestimated when blood samples were collected using reduviid bugs. In our validation experiment with nectar-feeding bats (Glossophaga soricina), we compared estimates of daily energy expenditure (DEE) using DLW with those derived from the energy balance method. We considered Speakman's equation (controlling for 25% fractionated water loss) as the most appropriate for our study animal and calculated DEE accordingly. On average, DEE estimated with DLW was not significantly different from the mean value obtained with the energy balance method (mean deviation 1.2%). We conclude that although bug hemolymph or intestinal liquids most likely contaminate the samples, estimates of DEE are still valid because the DLW method does not depend on absolute isotope enrichments but on the rate of isotope decrease over time. However, dilution of blood with intestinal liquids or hemolymph from a bug may lead to larger variation in DEE estimates. We also tested how the relative error of DLW estimates changed with varying assumptions about fractionation. We used three additional equations for calculating DEE in DLW experiments. The basic equation for DLW experiments published by Lifson and McClintock (LM-6) assumes no fractionation, resulted in an overestimate of DEE by 10%. Nagy's equation (N-2) controls for changes in body mass but not for

  18. Physiological and Pathological Impact of Blood Sampling by Retro-Bulbar Sinus Puncture and Facial Vein Phlebotomy in Laboratory Mice

    PubMed Central

    Holst, Birgitte; Hau, Jann; Rozell, Björn; Abelson, Klas Stig Peter

    2014-01-01

    Retro-bulbar sinus puncture and facial vein phlebotomy are two widely used methods for blood sampling in laboratory mice. However, the animal welfare implications associated with these techniques are currently debated, and the possible physiological and pathological implications of blood sampling using these methods have been sparsely investigated. Therefore, this study was conducted to assess and compare the impacts of blood sampling by retro-bulbar sinus puncture and facial vein phlebotomy. Blood was obtained from either the retro-bulbar sinus or the facial vein from male C57BL/6J mice at two time points, and the samples were analyzed for plasma corticosterone. Body weights were measured at the day of blood sampling and the day after blood sampling, and the food consumption was recorded automatically during the 24 hours post-procedure. At the end of study, cheeks and orbital regions were collected for histopathological analysis to assess the degree of tissue trauma. Mice subjected to facial vein phlebotomy had significantly elevated plasma corticosterone levels at both time points in contrast to mice subjected to retro-bulbar sinus puncture, which did not. Both groups of sampled mice lost weight following blood sampling, but the body weight loss was higher in mice subjected to facial vein phlebotomy. The food consumption was not significantly different between the two groups. At gross necropsy, subcutaneous hematomas were found in both groups and the histopathological analyses revealed extensive tissue trauma after both facial vein phlebotomy and retro-bulbar sinus puncture. This study demonstrates that both blood sampling methods have a considerable impact on the animals' physiological condition, which should be considered whenever blood samples are obtained. PMID:25426941

  19. Evaluation of blood microsampling techniques and sampling sites for the analysis of drugs by HPLC-MS.

    PubMed

    Smith, Christopher; Sykes, Angela; Robinson, Sally; Thomas, Elizabeth

    2011-01-01

    the potential for whole blood sampling (20 µl) in toxicokinetic studies to reduce the sample volume was investigated. Blood microsamples were collected in three ways, either as a dried blood spot (DBS), a blood sample collected in a micropipette placed in a plastic tube and mixed with water or as plasma in the normal manner. blood samples on the DBS and the whole blood microtube (WBMT) were compared along with DBS and plasma to determine the toxicokinetic data equivalency. The DBS and WBMT comparison was shown to be equivalent, as demonstrated on a correlation plot with an R(2) value of 0.97 for an x = y plot. The plasma comparison with DBS also gave a good correlation. A correction factor (x(2)) was applied to the blood data to allow for the distribution of the compound between plasma and bloods, and therefore, a direct comparison could be made. The correlation plot derived from the sample data gave an R(2) value 0.98 (x = y plot), indicating dataset equivalency. Sampling sites were evaluated in a dog study. Blood was collected from the peripheral region, in this case the ear, and a venous region of the dog; and spotted onto DBS cards. Comparison of the mean area under the curve data for the sampling sites showed equivalent data: 5095 and 5175 ng.h/ml for the 25 mg/kg dose and 16695 ng.h/ml and 16000 ng.h/ml for the 50 mg/kg dose for the ear and the venous samples, respectively. the DBS cards were shown to be an equivalent microsampling process when compared with WBMT and conventional plasma analysis. With the added benefits of sample storage, shipment and ease of use for DBS, this technology could change the way samples are taken and then analyzed in bioanalysis in the future.

  20. A round robin approach to the analysis of bisphenol a (BPA) in human blood samples

    PubMed Central

    2014-01-01

    Background Human exposure to bisphenol A (BPA) is ubiquitous, yet there are concerns about whether BPA can be measured in human blood. This Round Robin was designed to address this concern through three goals: 1) to identify collection materials, reagents and detection apparatuses that do not contribute BPA to serum; 2) to identify sensitive and precise methods to accurately measure unconjugated BPA (uBPA) and BPA-glucuronide (BPA-G), a metabolite, in serum; and 3) to evaluate whether inadvertent hydrolysis of BPA-G occurs during sample handling and processing. Methods Four laboratories participated in this Round Robin. Laboratories screened materials to identify BPA contamination in collection and analysis materials. Serum was spiked with concentrations of uBPA and/or BPA-G ranging from 0.09-19.5 (uBPA) and 0.5-32 (BPA-G) ng/mL. Additional samples were preserved unspiked as ‘environmental’ samples. Blinded samples were provided to laboratories that used LC/MSMS to simultaneously quantify uBPA and BPA-G. To determine whether inadvertent hydrolysis of BPA metabolites occurred, samples spiked with only BPA-G were analyzed for the presence of uBPA. Finally, three laboratories compared direct and indirect methods of quantifying BPA-G. Results We identified collection materials and reagents that did not introduce BPA contamination. In the blinded spiked sample analysis, all laboratories were able to distinguish low from high values of uBPA and BPA-G, for the whole spiked sample range and for those samples spiked with the three lowest concentrations (0.5-3.1 ng/ml). By completion of the Round Robin, three laboratories had verified methods for the analysis of uBPA and two verified for the analysis of BPA-G (verification determined by: 4 of 5 samples within 20% of spiked concentrations). In the analysis of BPA-G only spiked samples, all laboratories reported BPA-G was the majority of BPA detected (92.2 – 100%). Finally, laboratories were more likely to be verified

  1. Screening for lead poisoning in urban school children of southern India using capillary and venous blood samples.

    PubMed

    D'Souza, Herman S; Menezes, Geraldine; Venkatesh, T

    2002-01-01

    Our study aimed at comparing lead and zinc protoporphyrin (ZPP) levels in capillary and venous blood samples in a small population and to employ an easier method of sample collection for a major screening program in school children in major Indian cities. An awareness program on lead and its effects was conducted in two different schools. A total of thirty urban school children from South India, with an age group between 4-12 years consented for dual blood sampling and reported for the study. Venous and capillary blood samples were obtained simultaneously. Blood lead and zinc protoporphyrin (ZPP) levels were estimated using ESA Lead Analyzer and Haematofluorometer respectively. A significant correlation between capillary and venous ZPP (r=0.98) and lead (r=0.99) was observed. Rank sum test showed that there is no statistically significant difference between capillary and venous ZPP (P=0.891) and lead (P=0.672) values. This pilot study recommends that screening for lead may be done using capillary blood samples since significant correlation is observed between capillary and venous blood measurements. Obtaining samples using this mode is a non-invasive, less expensive, quick and easy method in children. Appropriately performed capillary sampling may be considered as an acceptable alternative to venipuncture for screening of blood for lead poisoning both in children and adults.

  2. Concentrations of persistent organic pollutants (POPs) in human blood samples from Mexico City, Mexico.

    PubMed

    Orta-García, Sandra; Pérez-Vázquez, Francisco; González-Vega, Carolina; Varela-Silva, José Antonio; Hernández-González, Lidia; Pérez-Maldonado, Iván

    2014-02-15

    Studies in Mexico have demonstrated exposure to persistent organic pollutants (POPs) in people living in different sites through the country. However, studies evaluating exposure to POPs in people living in Mexico City (one of most contaminated places in the world) are scarce. Therefore, the aim of this study was to assess the levels of polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolite dichlorodiphenyldichloroethylene (DDE) in the blood as exposure biomarkers in people living in Mexico City. A total of 123 participants (blood donors aged 20-60 years) were recruited during 2010 in Mexico City. Quantitative analyses of blood samples were performed using gas chromatography coupled with mass spectrometry. Levels of the assessed compounds ranged from non-detectable (

  3. Identification of food-derived bioactive peptides in blood and other biological samples.

    PubMed

    Sato, Kenji; Iwai, Koji; Aito-Inoue, Misako

    2008-01-01

    Recent studies have demonstrated the presence of food-derived peptides in human blood after ingestion of enzymatic hydrolysates of food proteins, while most peptides in food are degraded into amino acids during digestion and absorption. To capture and clarify the food-derived peptides in blood, solid-phase extraction (SPE) using a mini-spin column packed with a strong cation exchanger was developed. This technique allows the use of a nonvolatile acid such as trichloroacetic acid, a strong protein denaturant, for the deproteinizing procedure. To improve resolution of hydrophilic peptide and increase specificity and sensitivity in the detection of peptide by reversed-phase high-performance liquid chromatography (RP-HPLC) after subfractionation by size-exclusion chromatography (SEC), peptides are derivatized with phenyl isothiocyanate. The resultant phenyl thiocarbamyl (PTC)-peptides can be resolved with high resolution and sensitivity by RP-HPLC. By comparing chromatograms of PTC derivatives from blood before and after ingestion of a peptide sample, food-derived peptide can be detected. The isolated PTC-peptide can be applied to a peptide sequencer based on the Edman degradation reaction.

  4. Mitochondrial DNA levels in blood and tissue samples from breast cancer patients of different stages.

    PubMed

    Xia, Peng; Wang, Hui-Juan; Geng, Ting-Ting; Xun, Xiao-Jie; Zhou, Wen-Jing; Jin, Tian-Bo; Chen, Chao

    2014-01-01

    Alterations in mitochondrial DNA (mtDNA) have been implicated in carcinogenesis and tumor progression. We here evaluated the diagnostic and prognostic potential of mtDNA as a biomarker for breast cancer. Using multiplex real-time polymerase chain reaction, nuclear DNA (nDNA) and mtDNA levels in serum, buffy coat, tumor, and tumor-adjacent tissue samples from 50 breast cancer patients were determined and assessed for associations with clinicopathological features. To evaluate mtDNA as a biomarker for distinguishing between the four sample types, we created receiver operating characteristic (ROC) curves. The mtDNA levels in buffy coat were significantly lower than in other sample types. Relative to tumor-adjacent tissue, reduced levels of mtDNA were identified in buffy coat and tumor tissue but not in serum. According to ROC curve analysis, mtDNA levels could be used to distinguish between buffy coat and tumor-adjacent tissue samples with good sensitivity (77%) and specificity (83%). Moreover, mtDNA levels in serum and tumor tissue were positively associated with cancer TMN stage. The mtDNA levels in blood samples may represent a promising, non-invasive biomarker in breast cancer patients. Additional, large-scale validation studies are required to establish the potential use of mtDNA levels in the early diagnosis and monitoring of breast cancer.

  5. Cytokine assays: an assessment of the preparation and treatment of blood and tissue samples.

    PubMed

    Keustermans, Genoveva C E; Hoeks, Sanne B E; Meerding, Jenny M; Prakken, Berent J; de Jager, Wilco

    2013-05-15

    Cytokines are key components of the innate and adaptive immune system. As pivotal players in the progression or regression of a pathological process, these molecules provide a window through which diseases can be monitored and can thus act as biomarkers. In order to measure cytokine levels, a plethora of protocols can be applied. These methods include bioassays, protein microarrays, high-performance liquid chromatography (HPLC), sandwich enzyme-linked immunosorbent assay (ELISA), Meso Scale Discovery (MSD) electrochemiluminescence and bead based multiplex immunoassays (MIA). Due to the interaction and activity of cytokines, multiplex immunoassays are at the forefront of cytokine analysis by allowing multiple cytokines to be measured in parallel. However, even with optimized protocols, sample standardization needs to occur before these proteins can optimally act as biomarkers. This review describes various factors influencing the levels of cytokines measured in plasma, serum, dried blood spots and tissue biopsies, focusing on sample collection and handling, long term storage and the repetitive use of samples. By analyzing how each of these factors influences protein levels, it is concluded that samples should be stored at low temperatures in order to maintain cytokine stability. In addition, within a study, sample manipulations should be kept the same, with measurement protocols being chosen for their compatibility with the research in question. By having a clear understanding of what factors influence cytokine levels and how to overcome these technical issues, minimally confounded data can be obtained and cytokines can achieve optimal biomarker activity.

  6. [HLA antibodies in retroplacental blood samples. I. Preliminary results of a screening study].

    PubMed

    Rose, M; Waltz, H; Halle, H; Henske, B

    1982-01-01

    Retroplacental blood samples (between 3 ml and 290 ml) which had been obtained from 327 recently delivered women were tested for the presence of HLA-A, B, C, DR antibodies. The approaches taken by the examiners were the NIH microlymphocytotoxicity test according to Terasaki and the two-colour fluorescence test according to van Rood. The examiners established 66 sera with cytotoxic reactions, and 18 of these serum samples (5.5 per cent) contained good to very good antibodies of the HLA-A, B, C, and DR series (correlation coefficients being between 0.55 and 1.00). Advantages offered by this search test over conventional pregnancy screening are discussed in some detail.

  7. Slurry sampling in serum blood for mercury determination by CV-AFS.

    PubMed

    Aranda, Pedro R; Gil, Raúl A; Moyano, Susana; De Vito, Irma; Martinez, Luis D

    2009-01-30

    The heavy metal mercury (Hg) is a neurotoxin known to have a serious health impact even at relatively low concentrations. A slurry method was developed for the sensitive and precise determination of mercury in human serum blood samples by cold vapor generation coupled to atomic fluorescence spectrometry (CV-AFS). All variables related to the slurry formation were studied. The optimal hydrochloric concentration and tin(II) chloride concentration for CV generation were evaluated. Calibration within the range 0.1-10 microg L(-1) Hg was performed with the standard addition method, and compared with an external calibration. Additionally, the reliability of the results obtained was evaluated by analyzing mercury in the same samples, but submitted to microwave-assisted digestion method. The limit of detection was calculated as 25 ng L(-1) and the relative standard deviation was 3.9% at levels around of 0.4 microg L(-1)Hg.

  8. Performance characteristics of the HemoCue B-Glucose analyzer using whole-blood samples.

    PubMed

    Voss, E M; Cembrowski, G S

    1993-07-01

    We evaluated the HemoCue B-Glucose (HemoCue Inc, Mission Viejo, Calif) analyzer for accuracy, precision, linearity, and recovery. One hundred eighteen capillary whole-blood samples were analyzed in duplicate on the HemoCue B-Glucose and the YSI 2300 STAT Glucose/L-Lactate (Yellow Springs [Ohio] Instruments) analyzers; corresponding plasma glucose levels were measured in duplicate on the Roche Cobas MIRA (Roche Diagnostic Systems, Nutley, NJ) analyzer. Plasma glucose levels were converted to whole-blood equivalent glucose levels by using a factor of 1.11. The following regression equations were obtained: HemoCue = 1.02 (YSI) + 0.19, Sy/x = 0.52, r2 = .984; and HemoCue = 0.98 (whole-blood equivalent glucose levels) + 0.26, Sy/x = 0.55, r2 = .982. Within-run coefficients of variation were 4.0%, 3.5%, 2.2%, and 1.0% at glucose concentrations of 3.9, 5.4, 8.7, and 17.1 mmol/L (71, 97, 156, and 308 mg/dL), respectively. Between-run imprecision and total imprecision using lyopholized materials with three lot numbers of cuvettes were 4.2% and 5.6% at 2.1 mmol/L (37 mg/dL) and 2.4% and 2.7% at 5.2 mmol/L (95 mg/dL), respectively. The HemoCue B-Glucose analyzer displayed linearity between 0 and 22.2 mmol/L (0 and 400 mg/dL), and the percent recovery averaged 98.7% +/- 4.5% (mean +/- SD). The HemoCue B-Glucose analyzer is a rapid, simple, and reliable method for determinations of whole-blood glucose levels.

  9. A comparative evaluation of four DNA extraction protocols from whole blood sample.

    PubMed

    Ghaheri, M; Kahrizi, D; Yari, K; Babaie, A; Suthar, R S; Kazemi, E

    2016-03-31

    All organisms have Deoxyribonucleic acid (DNA) within their cells. DNA is a complex molecule that contains all of the information necessary to build and maintain an organism. DNA extraction is one of the most basic and essential techniques in the study of DNA that allow huge advances in molecular biology, biotechnology and bioinformatics laboratories. Whole blood samples are one of the main sources used to obtain DNA and there are many different protocols available in this issue. In current research, compared four DNA extraction protocols from blood samples; include modified phenol-chloroform protocol, two salting-out and enzyme free method and from commercial kit. The extracted DNAs by these protocols were analyzed according to their time demands, quality and quantity, toxicity and functionality in PCR method. Also the quality and quantity of the extracted DNA were surveyed by gel electrophoresis and Nanodrop spectrophotometry methods. It was observed that there are not significantly differences between these methods about DNA Purity (A260/A280), but the DNA yield (ng DNA/μl) of phenol/chloroform method was higher than other methods. In addition, phenol/chloroform was the most toxic method and it takes more time than other methods. Roche diagnostics GmbH kit was the most expensive among the four methods but the least extraction time was required and it was the safest method.

  10. Environmental contaminants in Texas, USA, wetland reptiles: Evaluation using blood samples

    USGS Publications Warehouse

    Clark, D.R.; Bickham, J.W.; Baker, D.L.; Cowman, D.F.

    2000-01-01

    Four species of reptiles (diamondback water snake [Nerodia rhombifer], blotched water snake [N. erythrogaster], cottonmouth [Agkistrodon piscivorus], and red-eared slider [Trachemys scripta]) were collected at two contaminated and three reference sites in Texas, USA. Old River Slough has received intensive applications of agricultural chemicals since the 1950s. Municipal Lake received industrial arsenic wastes continuously from 1940 to 1993. Blood samples were analyzed for organochlorines, potentially toxic elements, genetic damage, and plasma cholinesterase (ChE). Dichlorodiphenyldichloroethylene (DDE) concentrations reached as high as 3.0 ppm (wet weight) in whole blood of a diamondback water snake at Old River Slough, a level probably roughly equivalent to the maximum concentration found in plasma of peregrine falcons (Falco peregrinus) in 1978 to 1979 when DDE peaked in this sensitive species. Possible impacts on diamondback water snakes are unknown, but at least one diamondback water snake was gravid when captured, indicating active reproduction. Arsenic was not found in red-eared sliders (only species sampled) from Municipal Lake. Red-eared sliders of both sexes at Old River Slough showed declining levels of ChE with increasing mass, suggesting a life-long decrease of ChE levels. Possible negative population consequences are unknown, but no evidence was found in body condition (mass relative to carapace length) that red-eared sliders at either contaminated site were harmed.

  11. Methiopropamine in blood samples from drivers suspected of being under the influence of drugs.

    PubMed

    Tuv, Silja Skogstad; Bergh, Marianne Skov-Skov; Vindenes, Vigdis; Karinen, Ritva

    2016-01-01

    Methiopropamine (MPA; 1-(thiophen-2-yl)-2-methylaminopropane) belongs to the new psychoactive substances (NPS) that have emerged on the drug market in recent years. MPA appeared in 2011 and is an analogue of methamphetamine, sold as, for example, "Slush Eric" and "Blow." It is reported to have effects similar to those of methamphetamine, but the toxicity in humans is not known. Three fatal cases involving MPA have been reported. One analytical confirmed intoxication case has been published, and this supports the symptoms described by the users. The prevalence of recreational use of MPA is unknown, and no studies have reported the prevalence in driving under the influence of drug (DUID) cases. We investigated the frequency of MPA in DUID cases received at our institute during a 12-week period and report the analytical method using an ultraperformance liquid chromatography.tandem mass spectrometry for quantification of MPA in whole blood. The analytical findings were compared to the results from a clinical test of impairment performed by a physician shortly after the driving episode. The samples were analyzed for 42 different psychoactive substances. MPA was detected in 10 DUID cases (0.8% of the cases), only from male drivers. Other drugs were detected concomitantly in all the cases. Two of the cases were traffic accidents. Our study shows that MPA is found in DUID cases and reveals that NPS are used among drivers and also proven in blood from drivers involved in traffic accidents. More studies are requested regarding the pharmacological and toxicological effects of MPA and other NPS. This is the first article that describes a method for analyzing and quantifying MPA in whole blood samples.

  12. Evaluation of sample hemolysis in blood collected by S-Monovette using vacuum or aspiration mode.

    PubMed

    Lippi, Giuseppe; Avanzini, Paola; Musa, Roberta; Sandei, Franca; Aloe, Rosalia; Cervellin, Gianfranco

    2013-01-01

    In vitro hemolysis can be induced by several biological and technical sources, and may be worsened by forced aspiration of blood in vacuum tubes. This study was aimed to compare the probability of hemolysis by drawing blood with a commercial evacuated blood collection tube, and S-Monovette used either in the "vacuum" or "aspiration" mode. The study population consisted in 20 healthy volunteers. A sample was drawn into 4.0 mL BD Vacutainer serum tube from a vein of one upper arm. Two other samples were drawn with a second venipuncture from a vein of the opposite arm, into 4.0 mL S-Monovette serum tubes, by both vacuum an aspiration modes. After separation, serum potassium, lactate dehydrogenase (LD) and hemolysis index (HI) were tested on Beckman Coulter DxC. In no case the HI exceed the limit of significant hemolysis. As compared with BD Vacutainer, no significant differences were observed for potassium and LD using S-Monovette with vacuum method. Significant increased values of both parameters were however found in serum collected into BD Vacutainer and S-Monovette by vacuum mode, compared to serum drawn by S-Monovette in aspiration mode. The mean potassium bias was 2.2% versus BD Vacutainer and 2.4% versus S-Monovette in vacuum mode, that of LD was 2.7% versus BD Vacutainer and 2.1% versus S-Monovette in vacuum mode. None of these variations exceeded the allowable total error. Although no significant macro-hemolysis was observed with any collection system, the less chance of producing micro-hemolysis by S-Monovette in aspiration mode suggest that this device may be used when a difficult venipuncture combined with the vacuum may increase the probability of spurious hemolysis.

  13. Analysis of intraosseous blood samples using an EPOC point of care analyzer during resuscitation.

    PubMed

    Tallman, Crystal Ives; Darracq, Michael; Young, Megann

    2017-03-01

    In the early phases of resuscitation in a critically ill patient, especially those in cardiac arrest, intravenous (IV) access can be difficult to obtain. Intraosseous (IO) access is often used in these critical situations to allow medication administration. When no IV access is available, it is difficult to obtain blood for point of care analysis, yet this information can be crucial in directing the resuscitation. We hypothesized that IO samples may be used with a point of care device to obtain useful information when seconds really do matter. Patients presenting to the emergency department requiring resuscitation and IO placement were prospectively enrolled in a convenience sample. 17 patients were enrolled. IO and IV samples obtained within five minutes of one another were analyzed using separate EPOC® point of care analyzers. Analytes were compared using Bland Altman Plots and intraclass correlation coefficients. In this analysis of convenience sampled critically ill patients, the EPOC® point of care analyzer provided results from IO samples. IO and IV samples were most comparable for pH, bicarbonate, sodium and base excess, and potentially for lactic acid; single outliers for bicarbonate, sodium and base excess were observed. Intraclass correlation coefficients were excellent for sodium and reasonable for pH, pO2, bicarbonate, and glucose. Correlations for other variables measured by the EPOC® analyzer were not as robust. IO samples can be used with a bedside point of care analyzer to rapidly obtain certain laboratory information during resuscitations when IV access is difficult. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Comparison between dried blood spot and plasma sampling for therapeutic drug monitoring of antiepileptic drugs in children with epilepsy: A step towards home sampling.

    PubMed

    Linder, Camilla; Wide, Katarina; Walander, Malin; Beck, Olof; Gustafsson, Lars L; Pohanka, Anton

    2017-05-01

    To investigate if dried blood spots could be used for therapeutic drug monitoring of the antiepileptic drugs, carbamazepine, lamotrigine and valproic acid in children with epilepsy. Fingerprick blood samples from 46 children at a neuropediatric outpatient clinic was collected on filterpaper at the same time as capillary plasma sampling. A validated dried blood spot liquid chromatography tandem mass spectrometry method for carbamazepine, lamotrigine and valproic acid was compared with the routine plasma laboratory methods. Method agreement was evaluated and plasma concentrations were estimated by different conversion approaches. Strong correlation was shown between dried blood spot and plasma concentrations for all three drugs, with R2 values>0.89. Regression analysis showed a proportional bias with 35% lower dried blood spot concentrations for valproic acid (n=33) and concentrations were 18% higher for carbamazepine (n=17). A ratio approach was used to make a conversion from dried blood spots to estimated plasma for these two drugs. Dried blood spot concentrations were directly comparable with plasma for lamotrigine (n=20). This study supports that dried blood spot concentrations can be used as an alternative to plasma in a children population for three commonly used antiepileptic drugs with the possibility to expand by adding other antiepileptic drugs. Clinical decisions can be made based on converted (carbamazepine, valproic acid) or unconverted (lamotrigine) dried blood spot concentrations. Dried blood spot sampling, in the future taken at home, will simplify an effective therapeutic drug monitoring for this group of patients who often have concomitant disorders and also reduce costs for society. Copyright © 2016 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  15. Direct Trace Element Analysis of Liquid Blood Samples by In-Air Ion Beam Analytical Techniques (PIXE-PIGE).

    PubMed

    Huszank, Robert; Csedreki, László; Török, Zsófia

    2017-02-07

    There are various liquid materials whose elemental composition is of interest in various fields of science and technology. In many cases, sample preparation or the extraction can be complicated, or it would destroy the original environment before the analysis (for example, in the case of biological samples). However, multielement direct analysis of liquid samples can be realized by an external PIXE-PIGE measurement system. Particle-induced X-ray and gamma-ray emission spectroscopy (PIXE, PIGE) techniques were applied in external (in-air) microbeam configuration for the trace and main element determination of liquid samples. The direct analysis of standard solutions of several metal salts and human blood samples (whole blood, blood serum, blood plasma, and formed elements) was realized. From the blood samples, Na, P, S, Cl, K, Ca, Fe, Cu, Zn, and Br elemental concentrations were determined. The focused and scanned ion beam creates an opportunity to analyze very small volume samples (∼10 μL). As the sample matrix consists of light elements, the analysis is possible at ppm level. Using this external beam setup, it was found that it is possible to determine elemental composition of small-volume liquid samples routinely, while the liquid samples do not require any preparation processes, and thus, they can be analyzed directly. In the case of lower concentrations, the method is also suitable for the analysis (down to even ∼1 ppm level) but with less accuracy and longer measurement times.

  16. Clinical and anatomic pathology effects of serial blood sampling in rat toxicology studies, using conventional or microsampling methods.

    PubMed

    Caron, Alexis; Lelong, Christine; Bartels, T; Dorchies, O; Gury, T; Chalier, Catherine; Benning, Véronique

    2015-08-01

    As a general practice in rodent toxicology studies, satellite animals are used for toxicokinetic determinations, because of the potential impact of serial blood sampling on toxicological endpoints. Besides toxicological and toxicokinetic determinations, blood samples obtained longitudinally from a same animal may be used for the assessment of additional parameters (e.g., metabolism, pharmacodynamics, safety biomarkers) to maximize information that can be deduced from rodents. We investigated whether removal of up to 6 × 200 μL of blood over 24h can be applied in GLP rat toxicology studies without affecting the scientific outcome. 8 week-old female rats (200-300 g) were dosed for up to 1 month with a standard vehicle and subjected or not (controls) to serial blood sampling for sham toxicokinetic/ancillary determinations, using miniaturized methods allowing collection of 6 × 50, 100 or 200 μL over 24h. In-life endpoints, clinical pathology parameters and histopathology of organs sensitive to blood volume reduction were evaluated at several time points after completion of sampling. In sampled rats, minimal and reversible changes in red blood cell mass (maximally 15%) and subtle variations in liver enzymes, fibrinogen and neutrophils were not associated with any organ/tissue macroscopic or microscopic correlate. Serial blood sampling (up to 6 × 200 μL over 24h) is compatible with the assessment of standard toxicity endpoints in adult rats. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Blood

    MedlinePlus

    ... that die or are lost from the body. White Blood Cells White blood cells (WBCs, and also ... of severe pain. previous continue Diseases of the White Blood Cells Neutropenia (pronounced: new-truh-PEE-nee- ...

  18. Comparison of S. stercoralis Serology Performed on Dried Blood Spots and on Conventional Serum Samples

    PubMed Central

    Formenti, Fabio; Buonfrate, Dora; Prandi, Rosanna; Marquez, Monica; Caicedo, Cintia; Rizzi, Eleonora; Guevara, Angel G.; Vicuña, Yosselin; Huerlo, Francisco R.; Perandin, Francesca; Bisoffi, Zeno; Anselmi, Mariella

    2016-01-01

    Background: Dried blood spots (DBS) are used for epidemiological surveys on infectious diseases in settings where limited resources are available. In fact, DBS can help to overcome logistic difficulties for the collection, transport and storage of biological specimens. Objective: To evaluate the accuracy of Strongyloides stercoralis serology performed on DBS. Methods: A survey was proposed to children attending a school in the village of Borbon, Ecuador, and to their parents/guardians. Each participant gave consent to the collection of both serum and DBS specimens. DBS absorbed on filter papers were analyzed with a commercially available ELISA test for S. stercoralis antibodies, as well as with standard serology. The agreement between the two methods was assessed through the Cohen’s kappa coefficient. Results: The study sample was composed of 174 children and 61 adults, for a total of 235 serum and 235 DBS samples. The serology was positive in 31/235 (13%) serum samples, and in 27/235 (11%) DBS: 4 samples resulted discordant (positive at standard serology). Cohen’s kappa coefficient was 0.921 (95% CI 0.845 – 0.998), indicating a high rate of concordance. Conclusion: DBS are suitable for in field-surveys requiring serological testing for S. stercoralis. PMID:27877170

  19. Quantification of rifapentine, a potent antituberculosis drug, from dried blood spot samples using liquid chromatographic-tandem mass spectrometric analysis.

    PubMed

    Parsons, Teresa L; Marzinke, Mark A; Hoang, Thuy; Bliven-Sizemore, Erin; Weiner, Marc; Mac Kenzie, William R; Dorman, Susan E; Dooley, Kelly E

    2014-11-01

    The quantification of antituberculosis drug concentrations in multinational trials currently requires the collection of modest blood volumes, centrifugation, aliquoting of plasma, freezing, and keeping samples frozen during shipping. We prospectively enrolled healthy individuals into the Tuberculosis Trials Consortium Study 29B, a phase I dose escalation study of rifapentine, a rifamycin under evaluation in tuberculosis treatment trials. We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying rifapentine in whole blood on dried blood spots (DBS) to facilitate pharmacokinetic/pharmacodynamic analyses in clinical trials. Paired plasma and whole-blood samples were collected by venipuncture, and whole blood was spotted on Whatman protein saver 903 cards. The methods were optimized for plasma and then validated for DBS. The analytical measuring range for quantification of rifapentine and its metabolite was 50 to 80,000 ng/ml in whole-blood DBS. The analyte was stable on the cards for 11 weeks with a desiccant at room temperature and protected from light. The method concordance for paired plasma and whole-blood DBS samples was determined after correcting for participant hematocrit or population-based estimates of bias from Bland-Altman plots. The application of either correction factor resulted in acceptable correlation between plasma and whole-blood DBS (Passing-Bablok regression corrected for hematocrit; y = 0.98x + 356). Concentrations of rifapentine may be determined from whole-blood DBS collected via venipuncture after normalization in order to account for the dilutional effects of red blood cells. Additional studies are focused on the application of this methodology to capillary blood collected by finger stick. The simplicity of processing, storage, shipping, and low blood volume makes whole-blood DBS attractive for rifapentine pharmacokinetic evaluations, especially in international and pediatric trials.

  20. Quantification of Rifapentine, a Potent Antituberculosis Drug, from Dried Blood Spot Samples Using Liquid Chromatographic-Tandem Mass Spectrometric Analysis

    PubMed Central

    Parsons, Teresa L.; Marzinke, Mark A.; Hoang, Thuy; Bliven-Sizemore, Erin; Weiner, Marc; Mac Kenzie, William R.; Dorman, Susan E.

    2014-01-01

    The quantification of antituberculosis drug concentrations in multinational trials currently requires the collection of modest blood volumes, centrifugation, aliquoting of plasma, freezing, and keeping samples frozen during shipping. We prospectively enrolled healthy individuals into the Tuberculosis Trials Consortium Study 29B, a phase I dose escalation study of rifapentine, a rifamycin under evaluation in tuberculosis treatment trials. We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying rifapentine in whole blood on dried blood spots (DBS) to facilitate pharmacokinetic/pharmacodynamic analyses in clinical trials. Paired plasma and whole-blood samples were collected by venipuncture, and whole blood was spotted on Whatman protein saver 903 cards. The methods were optimized for plasma and then validated for DBS. The analytical measuring range for quantification of rifapentine and its metabolite was 50 to 80,000 ng/ml in whole-blood DBS. The analyte was stable on the cards for 11 weeks with a desiccant at room temperature and protected from light. The method concordance for paired plasma and whole-blood DBS samples was determined after correcting for participant hematocrit or population-based estimates of bias from Bland-Altman plots. The application of either correction factor resulted in acceptable correlation between plasma and whole-blood DBS (Passing-Bablok regression corrected for hematocrit; y = 0.98x + 356). Concentrations of rifapentine may be determined from whole-blood DBS collected via venipuncture after normalization in order to account for the dilutional effects of red blood cells. Additional studies are focused on the application of this methodology to capillary blood collected by finger stick. The simplicity of processing, storage, shipping, and low blood volume makes whole-blood DBS attractive for rifapentine pharmacokinetic evaluations, especially in international and pediatric trials. PMID:25182637

  1. Use of dried blood samples for monitoring hepatitis B virus infection

    PubMed Central

    Lira, Rosalia; Maldonado-Rodriguez, Angelica; Rojas-Montes, Othon; Ruiz-Tachiquin, Martha; Torres-Ibarra, Rocio; Cano-Dominguez, Carlos; Valdez-Salazar, Hilda; Gomez-Delgado, Alejandro; Muñoz, Onofre; Alvarez-Muñoz, Ma-Teresa

    2009-01-01

    Background Hepatitis B virus (HBV) infection is a problem in several regions of the world with limited resources. Blood samples dried on filter paper (DBS) have been successfully used to diagnose and monitor several infectious diseases. In Mexico there is an urgent need for an affordable and easy sampling method for viral load (VL) testing and monitoring of chronic HBV infection. The purpose of this work was to validate the utility of DBS samples for monitoring HBV infection in patients from Mexico City. Methods Matched samples of plasma and DBS on filter paper from 47 HBV infected patients from the Instituto Mexicano del Seguro Social (IMSS), were included. To evaluate the DNA stability and purity from DBS stored at different temperature conditions, samples from ten patients were stored at 4 degree, 25 degree, and 37 degree C for 7 days. After DBS elution and DNA extraction, the purity of these samples was determined measuring the O.D. rate 260/280. The DBS utility for molecular studies was assessed with PCR assays to amplify a 322 bp fragment from the "a" determinant region of the HBV "S" gene. The VL from all samples was determined to evaluate the correlation between plasma and DBS matched samples. Results The quality of the DNA from DBS specimen is not adversely affected by storage at 4 degree, 25 degree and 37 degree C for up 7 days. Statistical ANOVA analyses did not show any significant difference. The same amplification efficiency was observed between DNA templates from samples stored at different temperatures. The Pearson correlation between the VL from DBS and plasma matched samples was 0.93 (p = 0.01). The SD was 1.48 for DBS vs.1.32 for Plasma, and an average of log10 copies/mL of 5.32 vs. 5.53. ANOVA analysis did not show any statistically significant difference between the analyzed groups (p = 0.92). Conclusion The results provide strong evidence that the isolation and quantification of DNA-HBV from DBS is a viable alternative for patient monitoring

  2. Molecular identification and phylogenetic analysis of Wuchereria bancrofti from human blood samples in Egypt.

    PubMed

    Abdel-Shafi, Iman R; Shoieb, Eman Y; Attia, Samar S; Rubio, José M; Ta-Tang, Thuy-Huong; El-Badry, Ayman A

    2017-03-01

    Lymphatic filariasis (LF) is a serious vector-borne health problem, and Wuchereria bancrofti (W.b) is the major cause of LF worldwide and is focally endemic in Egypt. Identification of filarial infection using traditional morphologic and immunological criteria can be difficult and lead to misdiagnosis. The aim of the present study was molecular detection of W.b in residents in endemic areas in Egypt, sequence variance analysis, and phylogenetic analysis of W.b DNA. Collected blood samples from residents in filariasis endemic areas in five governorates were subjected to semi-nested PCR targeting repeated DNA sequence, for detection of W.b DNA. PCR products were sequenced; subsequently, a phylogenetic analysis of the obtained sequences was performed. Out of 300 blood samples, W.b DNA was identified in 48 (16%). Sequencing analysis confirmed PCR results identifying only W.b species. Sequence alignment and phylogenetic analysis indicated genetically distinct clusters of W.b among the study population. Study results demonstrated that the semi-nested PCR proved to be an effective diagnostic tool for accurate and rapid detection of W.b infections in nano-epidemics and is applicable for samples collected in the daytime as well as the night time. PCR products sequencing and phylogenitic analysis revealed three different nucleotide sequences variants. Further genetic studies of W.b in Egypt and other endemic areas are needed to distinguish related strains and the various ecological as well as drug effects exerted on them to support W.b elimination.

  3. Assessment of DDT and DDE levels in soil, dust, and blood samples from Chihuahua, Mexico.

    PubMed

    Martínez, Fernando Díaz-Barriga; Trejo-Acevedo, Antonio; Betanzos, Angel F; Espinosa-Reyes, Guillermo; Alegría-Torres, Jorge Alejandro; Maldonado, Iván Nelinho Pérez

    2012-02-01

    The aim of this study was to assess levels of DDT and DDE in two environmental matrices (soil and dust) and to investigate the blood levels of these insecticides in exposed children living in a north Mexican state (Chihuahua) where DDT was sprayed several years ago during (1) health campaigns for the control of malaria and (2) agricultural activities. DDT and DDE were analyzed by gas chromatography/mass spectrometry. In general, lower levels were found in household outdoor samples. The levels in outdoor samples ranged from 0.001 to 0.788 mg/kg for DDT and from 0.001 to 0.642 mg/kg for DDE. The levels in indoor samples ranged from 0.001 to 15.47 mg/kg for DDT and from 0.001 to 1.063 mg/kg for DDE. Similar results to those found in indoor soil were found in dust, in which the levels ranged from 0.001 to 95.87 mg/kg for DDT and from 0.001 to 0.797 mg/kg for DDE. Moreover, blood levels showed that all of the communities studied had been exposed to DDT and/or DDE, indicating a general past or present exposure to DDT. It is important to note that the quotient DDT/DDE in all matrices was always >1. Whether the people living in our study area are at risk is an issue that deserves further analysis. However, applying precautionary principles, it is important to initiate a risk-reduction program to decrease exposure to DDT and its metabolites in people living in this area.

  4. Cleaning the IceMole: collection of englacial samples from Blood Falls, Antarctica

    NASA Astrophysics Data System (ADS)

    Mikucki, J.; Digel, I.; Chua, M.; Davis, J.; Ghosh, D.; Lyons, W. B.; Welch, K. A.; Purcell, A.; Francke, G.; Feldmann, M.; Espe, C.; Heinen, D.; Dachwald, B.; Kowalski, J.; Tulaczyk, S. M.

    2016-12-01

    The Minimally Invasive Direct Glacial Access project (MIDGE) used a maneuverable thermoelectric melting probe called the IceMole to collect the first englacial samples of brine from Blood Falls, Antarctica. In order to maintain the scientific integrity of samples collected and minimize impact to this specially protected ecosystem, microbial and chemical contamination of the IceMole needed to be minimized. Guidelines have been established for research in Antarctic subglacial systems by the scientific and regulatory community and have been detailed by the "Code of Conduct for the Exploration and Research of Subglacial Aquatic Environments" put forth by the Scientific Committee on Antarctic Research (SCAR) Action Group, and was submitted to the Antarctic Treaty System. This Code of Conduct (CoC) recognizes the ecological importance and pristine nature of subglacial habitats and recommends a path forward towards clean exploration. Similarly, the US and European space agencies (NASA and ESA) have detailed instrument preparation protocols for the exploration of icy worlds in our solar system for planetary protection. Given the synergistic aims of these two groups we have adopted protocols from both subglacial and space exploration approaches. Here we present our approach to cleaning the IceMole in the field and report on ability to reduce the bioload inherent on the melter. Specifically our protocol reduced the exterior bio-load by an order of magnitude, to levels common in most clean rooms, and 1-3 orders of magnitude below that of Taylor Glacier ice surrounding Blood Falls. Our results indicate that the collection of englacial samples for microbiological analysis is feasible with melting probes.

  5. Bridging the gap between sample collection and laboratory analysis: using dried blood spots to identify human exposure to chemical agents

    NASA Astrophysics Data System (ADS)

    Hamelin, Elizabeth I.; Blake, Thomas A.; Perez, Jonas W.; Crow, Brian S.; Shaner, Rebecca L.; Coleman, Rebecca M.; Johnson, Rudolph C.

    2016-05-01

    Public health response to large scale chemical emergencies presents logistical challenges for sample collection, transport, and analysis. Diagnostic methods used to identify and determine exposure to chemical warfare agents, toxins, and poisons traditionally involve blood collection by phlebotomists, cold transport of biomedical samples, and costly sample preparation techniques. Use of dried blood spots, which consist of dried blood on an FDA-approved substrate, can increase analyte stability, decrease infection hazard for those handling samples, greatly reduce the cost of shipping/storing samples by removing the need for refrigeration and cold chain transportation, and be self-prepared by potentially exposed individuals using a simple finger prick and blood spot compatible paper. Our laboratory has developed clinical assays to detect human exposures to nerve agents through the analysis of specific protein adducts and metabolites, for which a simple extraction from a dried blood spot is sufficient for removing matrix interferents and attaining sensitivities on par with traditional sampling methods. The use of dried blood spots can bridge the gap between the laboratory and the field allowing for large scale sample collection with minimal impact on hospital resources while maintaining sensitivity, specificity, traceability, and quality requirements for both clinical and forensic applications.

  6. Bridging the Gap between Sample Collection and Laboratory Analysis: Using Dried Blood Spots to Identify Human Exposure to Chemical Agents.

    PubMed

    Hamelin, Elizabeth I; Blake, Thomas A; Perez, Jonas W; Crow, Brian S; Shaner, Rebecca L; Coleman, Rebecca M; Johnson, Rudolph C

    2016-05-13

    Public health response to large scale chemical emergencies presents logistical challenges for sample collection, transport, and analysis. Diagnostic methods used to identify and determine exposure to chemical warfare agents, toxins, and poisons traditionally involve blood collection by phlebotomists, cold transport of biomedical samples, and costly sample preparation techniques. Use of dried blood spots, which consist of dried blood on an FDA-approved substrate, can increase analyte stability, decrease infection hazard for those handling samples, greatly reduce the cost of shipping/storing samples by removing the need for refrigeration and cold chain transportation, and be self-prepared by potentially exposed individuals using a simple finger prick and blood spot compatible paper. Our laboratory has developed clinical assays to detect human exposures to nerve agents through the analysis of specific protein adducts and metabolites, for which a simple extraction from a dried blood spot is sufficient for removing matrix interferents and attaining sensitivities on par with traditional sampling methods. The use of dried blood spots can bridge the gap between the laboratory and the field allowing for large scale sample collection with minimal impact on hospital resources while maintaining sensitivity, specificity, traceability, and quality requirements for both clinical and forensic applications.

  7. Fungal DNA detected in blood samples of patients who received contaminated methylprednisolone injections reveals increased complexity of causative agents.

    PubMed

    Zhao, Yanan; Armeanu, Emilian; DiVerniero, Richard; Lewis, Terri A; Dobson, Richard C; Kontoyiannis, Dimitrios P; Roilides, Emmanuel; Walsh, Thomas J; Perlin, David S

    2014-06-01

    Using Exserohilum rostratum-specific and panfungal real-time PCR, we studied 24 blood samples and 2 synovial fluid specimens from 20 patients with persistent or worsening pain following injections of contaminated methylprednisolone. Seven blood specimens from 6 patients were significantly positive for fungal DNA by panfungal PCR, with multiple fungal species identified.

  8. Precision and accuracy of two blood glucose meters: FreeStyle Flash versus One Touch Ultra.

    PubMed

    Rivers, Shannon M; Kane, Michael P; Bakst, Gary; Busch, Robert S; Hamilton, Robert A

    2006-08-01

    The precision and accuracy of two blood glucose meters were evaluated using finger and forearm blood samples. Duplicate blood glucose measurements on the same forearm and finger as venipuncture were performed with the FreeStyle Flash and the One Touch Ultra. Accuracy was assessed by error-grid analysis and the number of values within 10% of the laboratory reference value. Precision was determined by calculating the absolute mean percent differences in glucose values between the first and second fingers and forearm test results. Forearm testing success was defined as an accurate glucose reading obtained with one lance. A total of 100 patients completed the study; 93% had diabetes and 53% were female. Patients' mean +/- S.D. age was 63 +/- 12 years, and glucose measurements ranged from 69 to 354 mg/dL. All finger-stick samples fell within error-grid zones A and B; 72% and 57% of FreeStyle Flash and One Touch Ultra values fell within 10% of the laboratory reference values, respectively (p = 0.027). Forearm samples were successfully obtained in 99 and 74 patients using the FreeStyle Flash and One Touch Ultra (p < 0.001), with 64 and 36 samples, respectively, falling within 10% of the laboratory reference values (p = 0.035). There was no difference in meter precision. The FreeStyle Flash and the One Touch Ultra are precise glucose meters; however, the FreeStyle Flash was associated with greater accuracy. Success rates of forearm glucose sampling were significantly greater when the FreeStyle Flash meter was used.

  9. Quantitative analysis of trace chromium in blood samples. Combination of the advanced oxidation process with catalytic adsorptive stripping voltammetry.

    PubMed

    Yong, Li; Armstrong, Kristie C; Dansby-Sparks, Royce N; Carrington, Nathan A; Chambers, James Q; Xue, Zi-Ling

    2006-11-01

    A new method for pretreating blood samples for trace Cr analysis is described. The advanced oxidation process (AOP with H2O2 and 5.5-W UV irradiation for 60 min) is used to remove biological/organic species for subsequent analysis. Prior to the AOP pretreatment, acid (HNO3) is used at pH 3.0 to inhibit the enzyme catalase in the blood samples. Catalytic adsorptive stripping voltammetry at a bismuth film electrode gives a Cr concentration of 6.0 +/- 0.3 ppb in the blood samples. This concentration was confirmed by dry-ashing the blood samples and subsequent analysis by atomic absorption spectroscopy. This current method may be used to monitor chromium, a trace metal in humans, and the efficacy and safety of chromium supplements as adjuvant therapy for diabetes.

  10. Description of a new non-injectable connector to reduce the complications of arterial blood sampling.

    PubMed

    Mariyaselvam, M Z; Heij, R E; Laba, D; Richardson, J A; Hodges, E J; Maduakor, C A; Carter, J J; Young, P J

    2015-01-01

    Arterial cannulation is associated with complications including bacterial contamination, accidental intra-arterial injection and blood spillage. We performed a series of audits and experiments to gauge the potential for these, as well as assess the possible contribution of a new device, the Needle-Free Arterial Non-Injectable Connector (NIC), in reducing these risks. The NIC comprises a needle-free connector that prevents blood spillage and a one-way valve allowing aspiration only; once screwed onto the side port of a three-way tap, the device can only be removed with difficulty. We performed a clinical audit of arterial monitoring systems in our intensive care unit, which showed an incidence of bacterial colonisation of five in 86 (6%) three-way tap ports. We constructed a manikin simulation experiment of the management of acute bradycardia, in which trainee doctors were required to inject atropine intravenously. Ten of 15 (66%) doctors injected the drug into the three-way tap of the arterial monitoring system rather than into the intravenous cannula or the central venous catheter. In a laboratory study, we replicated the arterial blood sampling and flushing sequence from a three-way tap, with the syringes attached either directly to the three-way tap port or to a NIC attached to the port. The first (discard) syringe attached to the three-way tap was contaminated with bacteria. Bacterial growth was found in 17 of 20 (85%) downstream flushed samples (corresponding to the patient's circulation) when the three-way tap was accessed directly, compared to none of 20 accessed via the NIC (p < 0.0001). Growth was found on all of 20 (100%) ports accessed directly compared to none of 20 accessed via the NIC (p < 0.0001). The NIC effectively prevents bacteria from contaminating sampling lines. As its design also prevents accidental intra-arterial injection, we suggest that it can reduce complications of arterial monitoring. © 2014 The Association of Anaesthetists of

  11. Evaluation of two-step haemoglobin screening with HemoCue for blood donor qualification in mobile collection sites.

    PubMed

    Gómez-Simón, A; Plaza, E M; Torregrosa, J M; Ferrer-Marín, F; Sánchez-Guiu, I; Vicente, V; Lozano, M L; Rivera, J

    2014-11-01

    Inaccuracy of fingerstick haemoglobin compromises donor's health and losses blood donations. We evaluated the benefit of double haemoglobin screening with HemoCue. Blood donors underwent fingerstick screening by HemoCue and were driven for donation if capillary haemoglobin was within the regulatory range. Those failing were drawn venous blood and donated if their venous haemoglobin determined with HemoCue was acceptable. Of 276 605 donor clinic visits, 10 011 (3·6%) were assessed by two-step haemoglobin screening using HemoCue, because of low (n = 9444) or high (n = 567) capillary haemoglobin. Among these, 2561 (25·6%) were deemed eligible [recovered donations]. The recovery rate was 23·8% and 55·0% among donors presenting with low and high capillary haemoglobin, respectively. In both categories of attempted donations, capillary and venous haemoglobin with HemoCue correlated significantly in recovered donors (R(2)  ≈ 0·5-0·7) but not in deferred visits (R(2)  < 0·15). Venous haemoglobin with HemoCue and by haematological analyzer significantly correlated in all donations attempts (R(2)  ≈ 0·7). Donors presenting with low capillary haemoglobin showed small bias between capillary and venous haemoglobin by HemoCue (-2·4 ± 6·2 g/l), fingerstick haemoglobin and venous haemoglobin with counter (1·3 ± 7·3 g/l), and venous haemoglobin with HemoCue and counter (3·7 ± 3·9 g/l). This bias was slightly greater in donors with high capillary haemoglobin (-7·5 ± 7·8, 13·7 ± 7·5, and 6·2 ± 7·5, respectively). Double haemoglobin screening by HemoCue reached an accuracy of 87·3% for qualifying donors presenting with low fingerstick haemoglobin. Double haemoglobin measurement with HemoCue [fingerstick and venous blood if required] is feasible and allows a significant recovery of blood donations. © 2014 International Society of Blood Transfusion.

  12. Whole blood flow cytometry measurements of in vivo platelet activation in critically-Ill patients are influenced by variability in blood sampling techniques.

    PubMed

    Rondina, Matthew T; Grissom, Colin K; Men, Shaohua; Harris, Estelle S; Schwertz, Hansjorg; Zimmerman, Guy A; Weyrich, Andrew S

    2012-06-01

    Flow cytometry is often used to measure in vivo platelet activation in critically-ill patients. Variability in blood sampling techniques, which may confound these measurements, remains poorly characterized. Platelet activation was measured by flow cytometry performed on arterial and venous blood from 116 critically-ill patients. We determined how variability in vascular sampling site, processing times, and platelet counts influenced levels of platelet-monocyte aggregates (PMA), PAC-1 binding (for glycoprotein (GP) IIbIIIa), and P-selectin (P-SEL) expression. Levels of PMA, but not PAC-1 binding or P-SEL expression, were significantly affected by variability in vascular sampling site. Average PMA levels were approximately 60% higher in whole blood drawn from an arterial vessel compared to venous blood (16.2±1.8% vs. 10.7±1.2%, p<0.05). Levels of PMA in both arterial and venous blood increased significantly during ex vivo processing delays (1.7% increase for every 10 minute delay, p<0.05). In contrast, PAC-1 binding and P-SEL expression were unaffected by processing delays. Levels of PMA, but not PAC-1 binding or P-SEL expression, were correlated with platelet count quartiles (9.4±1.6% for the lowest quartile versus 15.4±1.6% for the highest quartile, p<0.05). In critically-ill patients, variability in vascular sampling site, processing times, and platelet counts influence levels of PMA, but not PAC-1 binding or P-SEL expression. These data demonstrate the need for rigorous adherence to blood sampling protocols, particularly when levels of PMA, which are most sensitive to variations in blood collection, are measured for detection of in vivo platelet activation. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. Clean Sampling of an Englacial Conduit at Blood Falls, Antarctica - Some Experimental and Numerical Results

    NASA Astrophysics Data System (ADS)

    Kowalski, Julia; Francke, Gero; Feldmann, Marco; Espe, Clemens; Heinen, Dirk; Digel, Ilya; Clemens, Joachim; Schüller, Kai; Mikucki, Jill; Tulaczyk, Slawek M.; Pettit, Erin; Berry Lyons, W.; Dachwald, Bernd

    2017-04-01

    There is significant interest in sampling subglacial environments for geochemical and microbiological studies, yet those environments are typically difficult to access. Existing ice-drilling technologies make it cumbersome to maintain microbiologically clean access for sample acquisition and environmental stewardship of potentially fragile subglacial aquatic ecosystems. With the "IceMole", a minimally invasive, maneuverable subsurface ice probe, we have developed a clean glacial exploration technology for in-situ analysis and sampling of glacial ice and sub- and englacial materials. Its design is based on combining melting and mechanical stabilization, using an ice screw at the tip of the melting head to maintain firm contact between the melting head and the ice. The IceMole can change its melting direction by differential heating of the melting head and optional side wall heaters. Downward, horizontal and upward melting, as well as curve driving and penetration of particulate-ladden layers has already been demonstrated in several field tests. This maneuverability of the IceMole also necessitates a sophisticated on-board navigation system, capable of autonomous operations. Therefore, between 2012 and 2014, a more advanced probe was developed as part of the "Enceladus Explorer" (EnEx) project. The EnEx-IceMole offers systems for accurate positioning, based on in-ice attitude determination, acoustic positioning, ultrasonic obstacle and target detection, which is all integrated through a high-level sensor fusion algorithm. In December 2014, the EnEx-IceMole was used for clean access into a unique subglacial aquatic environment at Blood Falls, Antarctica, where an englacial brine sample was successfully obtained after about 17 meters of oblique melting. Particular attention was paid to clean protocols for sampling for geochemical and microbiological analysis. In this contribution, we will describe the general technological approach of the IceMole and report on the

  14. Comparison of total nucleated cell measurements of UC blood samples using two hematology analyzers.

    PubMed

    Eichler, H; Seetharaman, S; Latta, M; Kurtz, Jw; Moroff, G

    2004-01-01

    The total nucleated cell (TNC) content of umbilical cord blood (UCB) units currently serves as the most important measure for determining suitability for transplantation. Hence it is important that TNC measurements are performed in an accurate manner. TNC content is evaluated routinely by hematology analyzers (HA) as WBC counts. The objective of the study was to compare TNC content utilizing two different HA, one utilizing an impedance channel and optical channel, and the other using only an optical channel. The HA utilized in this study used two different modes of operation for lysis, regular mode (RM) and extended lysis mode (ELM). Cell-Dyn 3200 (CD3.2) utilizes optical technology for WBC measurements, involving WBC optical count (WOC) and nuclear optical count (NOC), whereas the Cell-Dyn 3700 (CD3.7) utilizes both the impedance (WIC) and optical technology (WOC) for WBC measurements. TNC content was determined with 17 identical samples using CD3.2 in one laboratory and CD3.7 in the other laboratory. Cord blood samples processed to concentrate nucleated cells by either of the laboratories were sent by overnight courier and assays were performed on the same day by both laboratories. For CD3.7, the WOC values were consistently lower than the WIC using the regular mode, but showed no significant differences (P>0.05). The WIC and WOC values were comparable on using the ELM and RM. For CD3.2, WOC values using RM and NOC values using ELM showed no significant differences (P>0.05), even though the WOC measurement was lower than the NOC values for most samples. The best comparison of TNC measurement between the two HA could be achieved by comparing CD3.7-WIC with CD3.2-NOC values. The results were equivalent (P>0.05) and 12 of 17 samples had equal to or less than 10% difference (mean 9.5%). TNC measurements of UCB samples were essentially identical using the WIC channel of the Cell-Dyn 3700 and the NOC channel of the Cell-Dyn 3200.

  15. Concentrations of cyanide in blood samples of corpses after smoke inhalation of varying origin.

    PubMed

    Stoll, Simone; Roider, Gabriele; Keil, Wolfgang

    2017-01-01

    Cyanide (CN) blood concentration is hardly considered during routine when evaluating smoke gas intoxications and fire victims, although some inflammable materials release a considerable amount of hydrogen cyanide. CN can be significant for the capacity to act and can in the end even be the cause of death. Systematic data concerning the influence of different fire conditions, especially those of various inflammable materials, on the CN-blood concentration of deceased persons do not exist. This study measured the CN level in 92 blood samples of corpses. All persons concerned were found dead in connection with fires and/or smoke gases. At the same time, the carboxyhemoglobin (COHb) level was determined, and the corpses were examined to detect pharmaceutical substances, alcohol and drugs. Furthermore, we analysed autopsy findings and the investigation files to determine the inflammable materials and other circumstances of the fires. Due to the inflammable materials, the highest concentration of CN in the victims was found after enclosed-space fires (n = 45) and after motor-vehicle fires (n = 8). The CN levels in these two groups (n = 53) were in 47 % of the cases toxic and in 13 % of the cases lethal. In victims of charcoal grills (n = 17) and exhaust gases (n = 6), no or only traces of CN were found. Only one case of the self-immolations (n = 12) displayed a toxic CN level. The results show that CN can have considerable significance when evaluating action ability and cause of death with enclosed-space fires and with motor-vehicle fires.

  16. Continuous quality control of the blood sampling procedure using a structured observation scheme

    PubMed Central

    Seemann, Tine Lindberg; Nybo, Mads

    2016-01-01

    Introduction An observational study was conducted using a structured observation scheme to assess compliance with the local phlebotomy guideline, to identify necessary focus items, and to investigate whether adherence to the phlebotomy guideline improved. Materials and methods The questionnaire from the EFLM Working Group for the Preanalytical Phase was adapted to local procedures. A pilot study of three months duration was conducted. Based on this, corrective actions were implemented and a follow-up study was conducted. All phlebotomists at the Department of Clinical Biochemistry and Pharmacology were observed. Three blood collections by each phlebotomist were observed at each session conducted at the phlebotomy ward and the hospital wards, respectively. Error frequencies were calculated for the phlebotomy ward and the hospital wards and for the two study phases. Results A total of 126 blood drawings by 39 phlebotomists were observed in the pilot study, while 84 blood drawings by 34 phlebotomists were observed in the follow-up study. In the pilot study, the three major error items were hand hygiene (42% error), mixing of samples (22%), and order of draw (21%). Minor significant differences were found between the two settings. After focus on the major aspects, the follow-up study showed significant improvement for all three items at both settings (P < 0.01, P < 0.01, and P = 0.01, respectively). Conclusion Continuous quality control of the phlebotomy procedure revealed a number of items not conducted in compliance with the local phlebotomy guideline. It supported significant improvements in the adherence to the recommended phlebotomy procedures and facilitated documentation of the phlebotomy quality. PMID:27812302

  17. Fetal blood sampling in twin pregnancies. Prenatal diagnosis and management of 19 cases.

    PubMed

    Cox, W L; Forestier, F; Capella-Pavlovsky, M; Daffos, F

    1987-01-01

    Twin pregnancies pose particular problems in both prenatal diagnosis and obstetric management. We present 19 twin pregnancies that underwent fetal blood sampling (FBS). The indications were mostly similar to those for singleton pregnancies, with both fetuses being sampled. There was one indication specific to twin pregnancies; disseminated intravascular coagulation in the retained twin after the death-in-utero (DIU) of the other. In 5 cases, only 1 twin was sampled; in 2 because the second twin was female in the diagnosis of an X-linked disorder; in 1 because of technical failure, and in 2 the other twin had predeceased. Eight pregnancies continued after the FBS delivering 2 live, healthy infants, though 5 were delivered before 37 weeks of gestation. In 7 cases there was a discordance in the diagnosis between the twins. In 3 of these cases the affected fetus underwent selective termination by air embolism; in 2 cases the pregnancies were continued and the affected twin not resuscitated; 1 pregnancy is still in progress, and 1 patient had a non-medically supervised termination of both twins in another country. Two patients miscarried within a week of the FBS. Two patients had only 1 living twin at the time of FBS; 1 had a second DIU a month after the FBS and the other a neonatal death at 11 days of age in an infant with severe porencephaly. FBS is technically feasible for similar indications as for singleton pregnancies though discordance in diagnosis raises specific management problems.

  18. Toxic metals status in human blood and breast milk samples in an integrated steel plant environment in Central India.

    PubMed

    Sharma, Rajnikant; Pervez, Shamsh

    2005-02-01

    Owing to its unique nutritional and immunological characteristics, human milk is the most important food source for infants. Breast milk can, however, also be a pathway of maternal excretion of toxic elements. Selected toxic elements (As, Pb, Mn,a Hg and Cd) were determined in human breast milk and blood samples obtained from 120 subjects related to an integrated steel plant environment located in central India. Samples of breast milk and blood from subjects living outside the steel plant environment were also analyzed for comparative study. Higher levels of these toxic elements were found in blood samples as compared to breast milk samples. Plant workers showed the higher presence of these metals in their breast milk and blood samples compared to the residents of the area and the subjects living outside the industrial environment, respectively. Mn, Pb and Hg have shown a higher tendency to associate with blood and breast milk than As and Cd. The order of occurrence of these metals in blood and milk samples thus found is Mn > Pb > Hg > As > Cd.

  19. The gingival vein as a minimally traumatic site for multiple blood sampling in guinea pigs and hamsters.

    PubMed

    Rodrigues, Mariana Valotta; de Castro, Simone Oliveira; de Albuquerque, Cynthia Zaccanini; Mattaraia, Vânia Gomes de Moura; Santoro, Marcelo Larami

    2017-01-01

    Laboratory animals are still necessary in scientific investigation and vaccine testing, but while novel methodological approaches are not available for their replacement, the search for new, humane, easy, and painless methods is necessary to diminish their stress and pain. When multiple blood samples are to be collected from hamsters and guinea pigs, the number of available venipuncture sites-which are greatly diminished in these species in comparison with other rodents due to the absence of a long tail-, harasses animal caregivers and researchers. Thus, this study aimed to evaluate if gingival vein puncture could be used as an additional route to obtain multiple blood samples from anesthetized hamsters and guinea pigs in such a way that animal behavior, well-being or hematological parameters would not be altered. Thus, twelve anesthetized Syrian golden hamsters and English guinea pigs were randomly allocated in two groups: a control group, whose blood samples were not collected, and an experimental group in which blood samples (200 microliters) were collected by gingival vein puncture at weekly intervals over six weeks. Clinical assessment, body weight gain and complete blood cell count were evaluated weekly, and control and experimental animals were euthanized at week seven, when the mentolabial region was processed to histological analyses. Multiple blood sampling from the gingival vein evoked no clinical manifestations or alteration in the behavioral repertoire, nor a statistically significant difference in weight gain in both species. Guinea pigs showed no alteration in red blood cell, leukocyte or platelet parameters over time. Hamsters developed a characteristic pattern of age-related physiological changes, which were considered normal. Histological analyses showed no difference in morphological structures in the interdental gingiva, confirming that multiple blood sampling is barely traumatic. Thus, these results evidence that blood collection from multiple

  20. Electrochemical aptasensor for lung cancer-related protein detection in crude blood plasma samples.

    PubMed

    Zamay, Galina S; Zamay, Tatiana N; Kolovskii, Vasilii A; Shabanov, Alexandr V; Glazyrin, Yury E; Veprintsev, Dmitry V; Krat, Alexey V; Zamay, Sergey S; Kolovskaya, Olga S; Gargaun, Ana; Sokolov, Alexey E; Modestov, Andrey A; Artyukhov, Ivan P; Chesnokov, Nikolay V; Petrova, Marina M; Berezovski, Maxim V; Zamay, Anna S

    2016-10-03

    The development of an aptamer-based electrochemical sensor for lung cancer detection is presented in this work. A highly specific DNA-aptamer, LC-18, selected to postoperative lung cancer tissues was immobilized onto a gold microelectrode and electrochemical measurements were performed in a solution containing the redox marker ferrocyanide/ferricyanide. The aptamer protein targets were harvested from blood plasma of lung cancer patients by using streptavidin paramagnetic beads and square wave voltammetry of the samples was performed at various concentrations. In order to enhance the sensitivity of the aptasensor, silica-coated iron oxide magnetic beads grafted with hydrophobic C8 and C4 alkyl groups were used in a sandwich detection approach. Addition of hydrophobic beads increased the detection limit by 100 times. The detection limit of the LC-18 aptasensor was enhanced by the beads to 0.023 ng/mL. The formation of the aptamer - protein - bead sandwich on the electrode surface was visualized by electron microcopy. As a result, the electrochemical aptasensor was able to detect cancer-related targets in crude blood plasma of lung cancer patients.

  1. Haematospirillum jordaniae gen. nov., sp. nov., isolated from human blood samples.

    PubMed

    Humrighouse, B W; Emery, B D; Kelly, A J; Metcalfe, M G; Mbizo, J; McQuiston, J R

    2016-04-01

    A Gram-negative, aerobic, motile, spiral-shaped bacterium, strain H5569(T), was isolated from a human blood sample. Phenotypic and molecular characteristics of the isolate were investigated. Optimal growth was found to occur at 35 °C under aerobic conditions on Heart Infusion Agar supplemented with 5 % rabbit blood. The major fatty acids present in the cells were identified as C16:0, C16:1ω7c and C18:1ω7c. The predominant respiratory quinone was found to be ubiquinone-Q10. The G+C content of genomic DNA for strain H5569(T) was found to be 49.9 %. Based on 16S rRNA gene sequence analysis results, 13 additional isolates were also analysed in this study. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the organism, represented by strain H5569(T), forms a distinct lineage within the family Rhodospirillaceae, closely related to two Novispirillum itersonii subspecies (93.9-94.1 %) and two Caenispirillum sp. (91.2-91.6 %). Based on these results, the isolate H5569(T) is concluded to represent a new genus and species for which the name Haematospirillum jordaniae gen. nov., sp. nov. is proposed. The type strain is H5569(T) (=DSM(T) 28903 = CCUG 66838(T)).

  2. Electrochemical aptasensor for lung cancer-related protein detection in crude blood plasma samples

    PubMed Central

    Zamay, Galina S.; Zamay, Tatiana N.; Kolovskii, Vasilii A.; Shabanov, Alexandr V.; Glazyrin, Yury E.; Veprintsev, Dmitry V.; Krat, Alexey V.; Zamay, Sergey S.; Kolovskaya, Olga S.; Gargaun, Ana; Sokolov, Alexey E.; Modestov, Andrey A.; Artyukhov, Ivan P.; Chesnokov, Nikolay V.; Petrova, Marina M.; Berezovski, Maxim V.; Zamay, Anna S.

    2016-01-01

    The development of an aptamer-based electrochemical sensor for lung cancer detection is presented in this work. A highly specific DNA-aptamer, LC-18, selected to postoperative lung cancer tissues was immobilized onto a gold microelectrode and electrochemical measurements were performed in a solution containing the redox marker ferrocyanide/ferricyanide. The aptamer protein targets were harvested from blood plasma of lung cancer patients by using streptavidin paramagnetic beads and square wave voltammetry of the samples was performed at various concentrations. In order to enhance the sensitivity of the aptasensor, silica-coated iron oxide magnetic beads grafted with hydrophobic C8 and C4 alkyl groups were used in a sandwich detection approach. Addition of hydrophobic beads increased the detection limit by 100 times. The detection limit of the LC-18 aptasensor was enhanced by the beads to 0.023 ng/mL. The formation of the aptamer – protein – bead sandwich on the electrode surface was visualized by electron microcopy. As a result, the electrochemical aptasensor was able to detect cancer-related targets in crude blood plasma of lung cancer patients. PMID:27694916

  3. Determination of lead in paired samples of human blood and synovial fluid

    SciTech Connect

    Villegas-Navarro, A.; Rosales, D.; Bustos, E.; Reyes, R.; Reyes, J.L.; Dieck, T.A.; Heredia, A. )

    1992-09-01

    In spite of the numerous papers published on the toxicity of lead in mammals, little is known about its effects in synovial fluid and bone joints. Our literature search showed a lack of quantitative studies regarding the concentration of lead in human synovial fluid; in addition, normal values regarding the threshold for poisoning by lead in that fluid are unknown. The available literature published corresponds to samples of human wounds by lead bullets localized close to or in a joint. Some of those papers dealing with lead-induced arthritis include symptoms of plumbism. They clearly demonstrate the ability of synovial fluid to dissolve lead and thereby make it available for systemic absorption. The molecular mechanism whereby this process is performed is still unknown, although it would be of interest because of its possible relationship with joint pain, a common problem in patients with lead poisoning that so far has not been fully explained. In a series of experiments with cattle, we found an average ratio of lead between synovial fluid and blood for paired observations of 4.2, although we have not found similar reports, and there is not sufficient information to make a total interpretation of these data. The purpose of this study was to determine the concentration of lead in synovial fluid and blood of corpses and to establish a possible numerical relationship between those two variables. 19 refs., 1 fig., 1 tab.

  4. Triclosan/triclocarban levels in maternal and umbilical blood samples and their association with fetal malformation.

    PubMed

    Wei, Ling; Qiao, Pengyun; Shi, Ying; Ruan, Yan; Yin, Jie; Wu, Qingqing; Shao, Bing

    2017-03-01

    Triclosan (TCS) and triclocarban (TCC) are widely used as antimicrobial compounds in consumer products. TCS and TCC are frequently found in waste water and sewage. In this study, we investigate the potential impact of exposure to triclosan (TCS) and triclocarban (TCC) on fetal abnormalities. We measured TCS and TCC levels in maternal and umbilical cord blood samples from 39 pregnant women diagnosed with fetal or post-birth abnormalities at Beijing Obstetrics and Gynecology Hospital. 52 pregnant women who gave birth to healthy neonates during the same period of time were included as controls. Applying ultra-performance liquid chromatography-tandem mass spectrometry, TCS and TCC concentrations were measured in maternal and fetal sera. Significantly increased levels of TCS were detected in maternal sera from mothers with abnormal births. Similar levels of TCS or TCC were found in maternal and cord sera in control group. The concentrations of TCS or TCC in maternal sera correlated with those in umbilical cord sera (r=0.649, P<0.01). These observations suggest that maternal blood test could be a useful assay for detecting fetal exposure to TCS and TCC, and high exposure to TCS may be potentially associated with increased risk for fetal malformations. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Antidepressants detection and quantification in whole blood samples by GC-MS/MS, for forensic purposes.

    PubMed

    Truta, Liliana; Castro, André L; Tarelho, Sónia; Costa, Pedro; Sales, M Goreti F; Teixeira, Helena M

    2016-09-05

    Depression is among the most prevalent psychiatric disorders of our society, leading to an increase in antidepressant drug consumption that needs to be accurately determined in whole blood samples in Forensic Toxicology Laboratories. For this purpose, this work presents a new gas chromatography tandem mass spectrometry (GC-MS/MS) method targeting the simultaneous and rapid determination of 14 common Antidepressants in whole blood: 13 Antidepressants (amitriptyline, citalopram, clomipramine, dothiepin, fluoxetine, imipramine, mianserin, mirtazapine, nortryptiline, paroxetine, sertraline, trimipramine and venlafaxine) and 1 Metabolite (N-desmethylclomipramine). Solid-phase extraction was used prior to chromatographic separation. Chromatographic and MS/MS parameters were selected to improve sensitivity, peak resolution and unequivocal identification of the eluted analyte. The detection was performed on a triple quadrupole tandem MS in selected ion monitoring (SIM) mode in tandem, using electronic impact ionization. Clomipramine-D3 and trimipramine-D3 were used as deutered internal standards. The validation parameters included linearity, limits of detection, lower limit of quantification, selectivity/specificity, extraction efficiency, carry-over, precision and robustness, and followed internationally accepted guidelines. Limits of quantification and detection were lower than therapeutic and sub-therapeutic concentration ranges. Overall, the method offered good selectivity, robustness and quick response (<16min) for typical concentration ranges, both for therapeutic and lethal levels. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Putative Epimutagens in Maternal Peripheral and Cord Blood Samples Identified Using Human Induced Pluripotent Stem Cells

    PubMed Central

    Arai, Yoshikazu; Hayakawa, Koji; Arai, Daisuke; Ito, Rie; Iwasaki, Yusuke; Saito, Koichi; Akutsu, Kazuhiko; Takatori, Satoshi; Ishii, Rie; Hayashi, Rumiko; Izumi, Shun-Ichiro; Sugino, Norihiro; Kondo, Fumio; Horie, Masakazu; Nakazawa, Hiroyuki; Makino, Tsunehisa; Hirosawa, Mitsuko; Shiota, Kunio; Ohgane, Jun

    2015-01-01

    The regulation of transcription and genome stability by epigenetic systems are crucial for the proper development of mammalian embryos. Chemicals that disturb epigenetic systems are termed epimutagens. We previously performed chemical screening that focused on heterochromatin formation and DNA methylation status in mouse embryonic stem cells and identified five epimutagens: diethyl phosphate (DEP), mercury (Hg), cotinine, selenium (Se), and octachlorodipropyl ether (S-421). Here, we used human induced pluripotent stem cells (hiPSCs) to confirm the effects of 20 chemicals, including the five epimutagens, detected at low concentrations in maternal peripheral and cord blood samples. Of note, these individual chemicals did not exhibit epimutagenic activity in hiPSCs. However, because the fetal environment contains various chemicals, we evaluated the effects of combined exposure to chemicals (DEP, Hg, cotinine, Se, and S-421) on hiPSCs. The combined exposure caused a decrease in the number of heterochromatin signals and aberrant DNA methylation status at multiple gene loci in hiPSCs. The combined exposure also affected embryoid body formation and neural differentiation from hiPSCs. Therefore, DEP, Hg, cotinine, Se, and S-421 were defined as an “epimutagen combination” that is effective at low concentrations as detected in maternal peripheral and cord blood. PMID:26339649

  7. Identification of malaria infected red blood samples by digital holographic quantitative phase microscope

    NASA Astrophysics Data System (ADS)

    Patel, Nimit R.; Chhaniwal, Vani K.; Javidi, Bahram; Anand, Arun

    2015-07-01

    Development of devices for automatic identification of diseases is desired especially in developing countries. In the case of malaria, even today the gold standard is the inspection of chemically treated blood smears through a microscope. This requires a trained technician/microscopist to identify the cells in the field of view, with which the labeling chemicals gets attached. Bright field microscopes provide only low contrast 2D images of red blood cells and cell thickness distribution cannot be obtained. Quantitative phase contrast microscopes can provide both intensity and phase profiles of the cells under study. The phase information can be used to determine thickness profile of the cell. Since cell morphology is available, many parameters pertaining to the 3D shape of the cell can be computed. These parameters in turn could be used to decide about the state of health of the cell leading to disease diagnosis. Here the investigations done on digital holographic microscope, which provides quantitative phase images, for comparison of parameters obtained from the 3D shape profile of objects leading to identification of diseased samples is described.

  8. Real-time PCR-based identification of bacterial and fungal pathogens from blood samples.

    PubMed

    Mai, Madeleine; Müller, Iris; Maneg, Daniela; Lohr, Benedikt; Haecker, Achim; Haberhausen, Gerd; Hunfeld, Klaus-Peter

    2015-01-01

    Latest major contributions in the field of sepsis diagnostics result from advances in PCR technologies permitting new standards in speed and quality, given the fact that a timely diagnosis is the decisive factor to the survival of patients with bloodstream infections.Multiplex real-time PCR is a quantitative method for simultaneous amplification and detection of different targeted DNA molecules within hours. Nevertheless, various studies have shown a number of technical shortcomings as well as a high heterogeneity in sensitivity.The present method allows the standardized and rapid detection and identification of 25 common bacteria and fungi responsible for bloodstream infections from whole blood samples by using LightCycler(®) SeptiFast (LC-SF) test, based on real-time PCR.

  9. A policy of routine umbilical cord blood gas analysis decreased missing samples from high-risk births.

    PubMed

    Ahlberg, M; Elvander, C; Johansson, S; Cnattingius, S; Stephansson, O

    2017-01-01

    This study compared obstetric units practicing routine or selective umbilical cord blood gas analysis, with respect to the risk of missing samples in high-risk deliveries and in infants with birth asphyxia. This was a Swedish population-based cohort study that used register data for 155 235 deliveries of live singleton infants between 2008 and 2014. Risk ratios and 95% confidence intervals were calculated to estimate the association between routine and selective umbilical cord blood gas sampling strategies and the risk of missing samples. Selective sampling increased the risk ratios when routine sampling was used as the reference, with a value of 1.0, and these were significant in high-risk deliveries and birth asphyxia. The risk ratios for selective sampling were large-for-gestational age (9.07), preterm delivery at up to 36 weeks of gestation (8.24), small-for-gestational age (7.94), two or more foetal scalp blood samples (5.96), an Apgar score of less than seven at one minute (2.36), emergency Caesarean section (1.67) and instrumental vaginal delivery (1.24). Compared with routine sampling, selective umbilical cord blood gas sampling significantly increased the risks of missing samples in high-risk deliveries and in infants with birth asphyxia. ©2016 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

  10. [Comparison of the determination of cyclosporin-A in blood samples collected on filter paper and by the ordinary technique].

    PubMed

    Azevedo, L S; Manrique, R; Sabbaga, E

    1995-01-01

    Monitoring cyclosporin-A (CsA) blood levels is of utmost importance for the rational use of this drug. Although many centers perform transplants, in Brazil there are few laboratories able to measure CsA blood levels. Therefore making blood samples reach the laboratory emerged as a problem. Collection of blood on filter paper has been a technique used for a long time in special cases. PURPOSE--To confirm the usefulness of measuring CsA blood levels in blood samples collected on filter paper and in the usual way. METHOD--We studied twenty renal cadaver kidney recipients who were receiving CsA, azathioprine and prednisone. Ninety five blood samples were collected and divided into two aliquots. One of them was sent routinely to one laboratory to perform whole blood CsA measurements. From the other aliquot, 20 microliters were pipetted on filter paper. When dried they were mailed to the other laboratory, where, after elution, CsA was measured. In both cases radioimmunoassay with polyclonal antibody was used. RESULTS--Linear correlation between both measurements revealed r = 0.81 with no statistical difference. CONCLUSION--The technique showed to be useful in clinical practice. In countries with continental size, as Brazil, it may be very helpful.

  11. Sulfatide Analysis by Mass Spectrometry for Screening of Metachromatic Leukodystrophy in Dried Blood and Urine Samples

    PubMed Central

    Spacil, Zdenek; Kumar, Arun Babu; Liao, Hsuan-Chieh; Auray-Blais, Christiane; Stark, Samantha; Suhr, Teryn R.; Scott, C. Ronald; Turecek, Frantisek; Gelb, Michael H.

    2016-01-01

    BACKGROUND Metachromatic leukodystrophy (MLD) is an autosomal recessive disorder caused by deficiency in arylsulfatase A activity, leading to accumulation of sulfatide substrates. Diagnostic and monitoring procedures include demonstration of reduced arylsulfatase A activity in peripheral blood leukocytes or detection of sulfatides in urine. However, the development of a screening test is challenging because of instability of the enzyme in dried blood spots (DBS), the widespread occurrence of pseudodeficiency alleles, and the lack of available urine samples from newborn screening programs. METHODS We measured individual sulfatide profiles in DBS and dried urine spots (DUS) from MLD patients with LC-MS/MS to identify markers with the discriminatory power to differentiate affected individuals from controls. We also developed a method for converting all sulfatide molecular species into a single species, allowing quantification in positive-ion mode upon derivatization. RESULTS In DBS from MLD patients, we found up to 23.2-fold and 5.1-fold differences in total sulfatide concentrations for early- and late-onset MLD, respectively, compared with controls and pseudodeficiencies. Corresponding DUS revealed up to 164-fold and 78-fold differences for early- and late-onset MLD patient samples compared with controls. The use of sulfatides converted to a single species simplified the analysis and increased detection sensitivity in positive-ion mode, providing a second option for sulfatide analysis. CONCLUSIONS This study of sulfatides in DBS and DUS suggests the feasibility of the mass spectrometry method for newborn screening of MLD and sets the stage for a larger-scale newborn screening pilot study. PMID:26585924

  12. Vaccination and blood sampling acceptability during Ramadan fasting month: A cross-sectional study in Conakry, Guinea.

    PubMed

    Peiffer-Smadja, Nathan; Ouedraogo, Ramatou; D'Ortenzio, Eric; Cissé, Papa Ndiaga; Zeggani, Zahra; Beavogui, Abdoul Habib; Faye, Sylvain Landry; Le Marcis, Frédéric; Yazdanpanah, Yazdan; Nguyen, Vinh-Kim

    2017-05-02

    There are few data on the acceptability of vaccination or blood sampling during Ramadan fasting month in Muslim countries. This could impact vaccination campaigns, clinical trials or healthcare during Ramadan. Using a semi-structured questionnaire, we conducted a cross-sectional study on 201 practising Muslims and 10 religious leaders in Conakry, Guinea in the wake of the recent epidemic Ebola epidemic. Acceptability of vaccination and blood sampling during Ramadan were investigated as well as reasons for refusal. Vaccination was judged acceptable during Ramadan by 46% (93/201, 95% CI 0.40-0.53) of practising Muslims versus 80% (8/10, 95% CI 0.49-0.94) of religious leaders (p=0.11). Blood sampling was judged acceptable during Ramadan by 54% (108/201, 95% CI 0.47-0.60) of practising Muslims versus 80% (8/10, 95% CI 0.49-0.94) of religious leaders (p=0.19). The percentage of participants that judged both blood sampling and vaccination acceptable during Ramadan was 40% (81/201, 95% CI 0.34-0.47) for practising Muslims versus 80% (8/10, 95% CI 0.49-0.94) for religious leaders (p=0.048). The most common reasons for refusal of vaccination or blood sampling were that nothing should enter or leave the body during Ramadan (43%), that adverse events could lead to breaking the fast (32%), that blood should not be seen during Ramadan (9%) and that the Quran explicitly forbids it (9%). Although most Muslims leaders and scientists consider that injections including immunization and blood sampling should be authorized during Ramadan, many Muslims in our study judged vaccination or blood sampling unacceptable when fasting. Widely available recommendations on healthcare during Ramadan would be useful to inform Muslims. Copyright © 2017. Published by Elsevier Ltd.

  13. Preterm Cord Blood Contains a Higher Proportion of Immature Hematopoietic Progenitors Compared to Term Samples

    PubMed Central

    Podestà, Marina; Bruschettini, Matteo; Cossu, Claudia; Sabatini, Federica; Dagnino, Monica; Romantsik, Olga; Spaggiari, Grazia Maria; Ramenghi, Luca Antonio; Frassoni, Francesco

    2015-01-01

    Background Cord blood contains high number of hematopoietic cells that after birth disappear. In this paper we have studied the functional properties of the umbilical cord blood progenitor cells collected from term and preterm neonates to establish whether quantitative and/or qualitative differences exist between the two groups. Methods and Results Our results indicate that the percentage of total CD34+ cells was significantly higher in preterm infants compared to full term: 0.61% (range 0.15–4.8) vs 0.3% (0.032–2.23) p = 0.0001 and in neonates <32 weeks of gestational age (GA) compared to those ≥32 wks GA: 0.95% (range 0.18–4.8) and 0.36% (0.15–3.2) respectively p = 0.0025. The majority of CD34+ cells co-expressed CD71 antigen (p<0.05 preterm vs term) and grew in vitro large BFU-E, mostly in the second generation. The subpopulations CD34+CD38- and CD34+CD45- resulted more represented in preterm samples compared to term, conversely, Side Population (SP) did not show any difference between the two group. The absolute number of preterm colonies (CFCs/10microL) resulted higher compared to term (p = 0.004) and these progenitors were able to grow until the third generation maintaining an higher proportion of CD34+ cells (p = 0.0017). The number of colony also inversely correlated with the gestational age (Pearson r = -0.3001 p<0.0168). Conclusions We found no differences in the isolation and expansion capacity of Endothelial Colony Forming Cells (ECFCs) from cord blood of term and preterm neonates: both groups grew in vitro large number of endothelial cells until the third generation and showed a transitional phenotype between mesenchymal stem cells and endothelial progenitors (CD73, CD31, CD34 and CD144)The presence, in the cord blood of preterm babies, of high number of immature hematopoietic progenitors and endothelial/mesenchymal stem cells with high proliferative potential makes this tissue an important source of cells for developing new cells therapies

  14. Blood typing

    MedlinePlus

    A blood sample is needed. The test to determine your blood group is called ABO typing. Your blood sample is mixed with antibodies against type A and B blood. Then, the sample is checked to see whether ...

  15. Rapid isolation of viable circulating tumor cells from patient blood samples.

    PubMed

    Hughes, Andrew D; Mattison, Jeff; Powderly, John D; Greene, Bryan T; King, Michael R

    2012-06-15

    following isolation, and these samples typically consist of >50% viable primary cancer cells from each patient. This device has been used to capture viable CTC from both diluted whole blood and buffy coat samples. Ultimately, we present a technique with functionality in a clinical setting to develop personalized cancer therapies.

  16. Rapid Isolation of Viable Circulating Tumor Cells from Patient Blood Samples

    PubMed Central

    Hughes, Andrew D.; Mattison, Jeff; Powderly, John D.; Greene, Bryan T.; King, Michael R.

    2012-01-01

    following isolation, and these samples typically consist of >50% viable primary cancer cells from each patient. This device has been used to capture viable CTC from both diluted whole blood and buffy coat samples. Ultimately, we present a technique with functionality in a clinical setting to develop personalized cancer therapies. PMID:22733259

  17. Comparison of glucose determinations on blood samples collected in three types of tubes.

    PubMed

    Li, Geling; Cabanero, Michael; Wang, Zhenglong; Wang, Huiying; Huang, Tao; Alexis, Herol; Eid, Ikram; Muth, Gilad; Pincus, Matthew R

    2013-01-01

    Because of the metabolism of serum glucose in collection tubes containing blood samples, serum glucose levels may be found to decrease over time. Several types of collection tubes have been designed to, at least partially, block glucose metabolism by red blood cells in blood collection tubes that may not be analyzed immediately after blood collection. These include red-top collection tubes with serum separator, grey-top tubes with a fluoride glycolysis inhibitor, and heparin-containing green-top tubes which prevent clot formation. As part of a quality assurance project, we investigated whether glucose levels differed in the three tube types from each of 18 volunteers on a prolonged standing of 4 hours. We then determined the glucose concentrations of all three tubes from each of the 18 volunteers. We used refrigerated samples over a five-day period to determine if the initial values were reproducible. Surprisingly, after standing for four hours at room temperature, we found that the glucose levels in the three tubes from each volunteer were statistically indistinguishable from one another using the two-tailed paired t-test. Also, a linear regression analysis showed that the values of glucose for the three pairs of two tube types were closely correlated with one another, with correlation coefficients of >0.97, slopes close to 1, and Y-intercepts close to 0. These results suggest that blood collection in any of these tubes will render similar values for serum glucose even after standing for four hours. The tubes were then refrigerated at 4°C and re-analyzed after another six hours and then once per day for the next four days. Beginning at the first day at the six-hour determination, the glucose levels in the red- and grey-top tubes were statistically indistinguishable from one another but not in the red- and green-top tubes and in the grey- and green-top tubes. This was due to a steady decrease in the glucose levels in the green-top tubes. The glucose levels in the

  18. Using Dried Blood Spot Sampling to Improve Data Quality and Reduce Animal Use in Mouse Pharmacokinetic Studies

    PubMed Central

    Wickremsinhe, Enaksha R; Perkins, Everett J

    2015-01-01

    Traditional pharmacokinetic analysis in nonclinical studies is based on the concentration of a test compound in plasma and requires approximately 100 to 200 µL blood collected per time point. However, the total blood volume of mice limits the number of samples that can be collected from an individual animal—often to a single collection per mouse—thus necessitating dosing multiple mice to generate a pharmacokinetic profile in a sparse-sampling design. Compared with traditional methods, dried blood spot (DBS) analysis requires smaller volumes of blood (15 to 20 µL), thus supporting serial blood sampling and the generation of a complete pharmacokinetic profile from a single mouse. Here we compare plasma-derived data with DBS-derived data, explain how to adopt DBS sampling to support discovery mouse studies, and describe how to generate pharmacokinetic and pharmacodynamic data from a single mouse. Executing novel study designs that use DBS enhances the ability to identify and streamline better drug candidates during drug discovery. Implementing DBS sampling can reduce the number of mice needed in a drug discovery program. In addition, the simplicity of DBS sampling and the smaller numbers of mice needed translate to decreased study costs. Overall, DBS sampling is consistent with 3Rs principles by achieving reductions in the number of animals used, decreased restraint-associated stress, improved data quality, direct comparison of interanimal variability, and the generation of multiple endpoints from a single study. PMID:25836959

  19. Capillary blood sampling as an alternative to venipuncture in the assessment of serum 25 hydroxyvitamin D levels.

    PubMed

    Dayre McNally, J; Matheson, Loren A; Sankaran, Koravangattu; Rosenberg, Alan M

    2008-11-01

    This study compared 25-hydroxyvitamin D [25(OH)D] measurements in capillary and venous blood samples collected, respectively by fingerprick and venipuncture. Capillary blood for measuring 25(OH)D has potential advantages by reducing blood volume required (2mL versus 0.3mL for venipuncture and capillary sampling, respectively), facilitating blood collection for those populations in whom venipuncture is difficult (e.g. infants and children), improving patient convenience and reducing costs associated with phlebotomy. The results demonstrated a highly significant relationship between 25(OH)D levels in serum derived from venous and capillary blood samples (r(2)=0.901). Despite statistically higher 25(OH)D levels in fingerprick samples (108+/-9nmol/L) compared with venipuncture samples (90+/-7nmol/L), the correlation between venous and capillary samples provides support for this approach as a practical alternative to venipuncture for vitamin D determination. However, clinical application may require the incorporation of a correction factor for the assessment of insufficiency, and research studies should avoid using the two methods interchangeably. Studying vitamin D's role in health and disease requires collection techniques and measurement methods that are reliable, reproducible, easily accessible, inexpensive and minimally burdensome to the patient. The option to collect patient samples by fingerprick may facilitate the collection process.

  20. Using dried blood spot sampling to improve data quality and reduce animal use in mouse pharmacokinetic studies.

    PubMed

    Wickremsinhe, Enaksha R; Perkins, Everett J

    2015-03-01

    Traditional pharmacokinetic analysis in nonclinical studies is based on the concentration of a test compound in plasma and requires approximately 100 to 200 μL blood collected per time point. However, the total blood volume of mice limits the number of samples that can be collected from an individual animal-often to a single collection per mouse-thus necessitating dosing multiple mice to generate a pharmacokinetic profile in a sparse-sampling design. Compared with traditional methods, dried blood spot (DBS) analysis requires smaller volumes of blood (15 to 20 μL), thus supporting serial blood sampling and the generation of a complete pharmacokinetic profile from a single mouse. Here we compare plasma-derived data with DBS-derived data, explain how to adopt DBS sampling to support discovery mouse studies, and describe how to generate pharmacokinetic and pharmacodynamic data from a single mouse. Executing novel study designs that use DBS enhances the ability to identify and streamline better drug candidates during drug discovery. Implementing DBS sampling can reduce the number of mice needed in a drug discovery program. In addition, the simplicity of DBS sampling and the smaller numbers of mice needed translate to decreased study costs. Overall, DBS sampling is consistent with 3Rs principles by achieving reductions in the number of animals used, decreased restraint-associated stress, improved data quality, direct comparison of interanimal variability, and the generation of multiple endpoints from a single study.

  1. Serum levels of perfluoroalkyl compounds in human maternal and umbilical cord blood samples

    SciTech Connect

    Monroy, Rocio; Morrison, Katherine; Teo, Koon; Atkinson, Stephanie; Kubwabo, Cariton; Stewart, Brian; Foster, Warren G.

    2008-09-15

    Perfluoroalkyl compounds (PFCs) are end-stage metabolic products from industrial flourochemicals used in the manufacture of plastics, textiles, and electronics that are widely distributed in the environment. The objective of the present study was to quantify exposure to perfluorooctane sulfonate (PFOS), perfluorooctanoate (PFOA), perfluorodecanoic acid (PFDeA), perfluorohexane sulfonate (PFHxS), perfluoroheptanoic acid (PFHpA), and perfluorononanoic acid (PFNA) in serum samples collected from pregnant women and the umbilical cord at delivery. Pregnant women (n=101) presenting for second trimester ultrasound were recruited and PFC residue levels were quantified in maternal serum at 24-28 weeks of pregnancy, at delivery, and in umbilical cord blood (UCB; n=105) by liquid chromatography-mass spectrometry. Paired t-test and multiple regression analysis were performed to determine the relationship between the concentrations of each analyte at different sample collection time points. PFOA and PFOS were detectable in all serum samples analyzed including the UCB. PFOS serum levels (mean{+-}S.D.) were significantly higher (p<0.001) in second trimester maternal serum (18.1{+-}10.9 ng/mL) than maternal serum levels at delivery (16.2{+-}10.4 ng/mL), which were higher than the levels found in UCB (7.3{+-}5.8 ng/mL; p<0.001). PFHxS was quantifiable in 46/101 (45.5%) maternal and 21/105 (20%) UCB samples with a mean concentration of 4.05{+-}12.3 and 5.05{+-}12.9 ng/mL, respectively. There was no association between serum PFCs at any time point studied and birth weight. Taken together our data demonstrate that although there is widespread exposure to PFCs during development, these exposures do not affect birth weight.

  2. Dried Blood Spot Sampling for Tacrolimus and Mycophenolic Acid in Children: Analytical and Clinical Validation.

    PubMed

    Martial, Lisa C; Hoogtanders, Karin E J; Schreuder, Michiel F; Cornelissen, Elisabeth A; van der Heijden, Jac; Joore, Manuela A; Van Maarseveen, Erik M; Burger, David M; Croes, Sander; Brüggemann, Roger J M; Aarnoutse, Rob E

    2017-08-01

    Tacrolimus and mycophenolic acid (MPA) are the backbone of immunosuppressive therapy after pediatric kidney transplantation. Dosing of these drugs is individualized by therapeutic drug monitoring. Dried blood spot (DBS) sampling may prove beneficial over conventional venous sampling. We aimed to develop and clinically validate a DBS method for tacrolimus and MPA in children. A joint DBS liquid chromatography-mass spectrometry assay for tacrolimus and MPA was developed. DBS-specific items included the hematocrit effect and influence of spot volume. Subsequently, a clinical validation study among children aged 2-18 years was performed to assess the agreement between observed and DBS-predicted venous concentrations. Agreement of the methods was assessed with Passing-Bablok regression, Bland-Altman plots, and quantification of the DBS predictive performance in terms of bias (median percentage prediction error) and precision (median absolute percentage prediction error), both should be <15%. A total of 40 tacrolimus and 32 MPA samples were available from 28 children. Conversion factors were used to predict venous concentrations from DBS. For tacrolimus, 95% of the individual ratios of predicted and observed concentrations were within a range of 0.74-1.28, with 85% of these ratios between 0.80 and 1.20 (Bland-Altman plots). For MPA, the 95% limits of agreement represented a broader range of 0.49-1.49%, and 72% of individual ratios were between the 0.80 and 1.20 limits. Median percentage prediction error and median absolute percentage prediction error were less than 15% for both drugs. A DBS assay was developed for tacrolimus and MPA. Tacrolimus venous concentrations could be adequately predicted from DBS. DBS analysis of MPA seemed to be a semiquantitative measurement at the most when compared with conventional plasma analysis, considering the high variability between observed and predicted concentrations. Next, home-based DBS sampling of tacrolimus for the purpose of

  3. Evaluation of occupational exposure in a slide bearings factory on the basis of urine and blood sample analyses.

    PubMed

    Raińska, Emilia; Biziuk, Marek; Jaremin, Bogdan; Głombiowski, Piotr; Fodor, Peter; Bielawski, Leszek

    2007-04-01

    The impact of a slide bearings factory on its workers was examined. Urine and blood samples were collected from 42 workers and six people employed in the offices in the same factory (control group). Concentrations of Al, Cu, Pb and Zn in blood and urine samples were measured twice (before and after chelation therapy) by ICP-MS technique using standard addition method. The essential differences in concentrations of elements for workers and control group were evaluated using non-parametric Mann-Whitney U-test. Significant differences between workers and control group were found for Pb in blood and Al in urine samples. The study was also undertaken to indicate correlation between blood and urine element content, workers' ages, their period of work and work section. It was also found that intravenous administration of 1 g of calcium-disodium versanate significantly increased urinary excretion of Pb and Zn, but not Al.

  4. Blood parameters are little affected by time of sampling after the application of ketamine in black howler monkeys (Alouatta pigra).

    PubMed

    Rovirosa-Hernández, M J; García-Orduña, F; Caba, M; Canales-Espinosa, D; Hermida-Lagunes, J; Torres-Pelayo, V R

    2011-10-01

    Ketamine hydrochloride is an anesthetic commonly utilized to obtain biological samples in various non-human primates. Its application alters individual hematologic and biochemical values. The aim of this study was to analyze its effect on blood parameters of Alouatta pigra. We collected blood samples at 10 and 40 minutes after the application of ketamine in 12 adult female A. pigra living in free-ranging conditions. The analysis showed that 40 minutes after application of ketamine, the number of platelets, lymphocytes and concentration of phosphorus decreased; however, creatinine, cholesterol, triglycerides, and potassium values increased. Our results suggest that ketamine appears to have little effect on the hematology and blood biochemistry of Alouatta pigra females with respect to those reported for other non-human primates. It is also important to consider the elapsed time after their application when taking blood samples for proper interpretation of the hemogram of Alouatta pigra females. © 2011 John Wiley & Sons A/S.

  5. Comparative Analysis of Clinical Samples Showing Weak Serum Reaction on AutoVue System Causing ABO Blood Typing Discrepancies.

    PubMed

    Jo, Su Yeon; Lee, Ju Mi; Kim, Hye Lim; Sin, Kyeong Hwa; Lee, Hyeon Ji; Chang, Chulhun Ludgerus; Kim, Hyung Hoi

    2017-03-01

    ABO blood typing in pre-transfusion testing is a major component of the high workload in blood banks that therefore requires automation. We often experienced discrepant results from an automated system, especially weak serum reactions. We evaluated the discrepant results by the reference manual method to confirm ABO blood typing. In total, 13,113 blood samples were tested with the AutoVue system; all samples were run in parallel with the reference manual method according to the laboratory protocol. The AutoVue system confirmed ABO blood typing of 12,816 samples (97.7%), and these results were concordant with those of the manual method. The remaining 297 samples (2.3%) showed discrepant results in the AutoVue system and were confirmed by the manual method. The discrepant results involved weak serum reactions (<2+ reaction grade), extra serum reactions, samples from patients who had received stem cell transplants, ABO subgroups, and specific system error messages. Among the 98 samples showing ≤1+ reaction grade in the AutoVue system, 70 samples (71.4%) showed a normal serum reaction (≥2+ reaction grade) with the manual method, and 28 samples (28.6%) showed weak serum reaction in both methods. ABO blood tying of 97.7% samples could be confirmed by the AutoVue system and a small proportion (2.3%) needed to be re-evaluated by the manual method. Samples with a 2+ reaction grade in serum typing do not need to be evaluated manually, while those with ≤1+ reaction grade do.

  6. Comparative Analysis of Clinical Samples Showing Weak Serum Reaction on AutoVue System Causing ABO Blood Typing Discrepancies

    PubMed Central

    Jo, Su Yeon; Lee, Ju Mi; Kim, Hye Lim; Sin, Kyeong Hwa; Lee, Hyeon Ji; Chang, Chulhun Ludgerus

    2017-01-01

    Background ABO blood typing in pre-transfusion testing is a major component of the high workload in blood banks that therefore requires automation. We often experienced discrepant results from an automated system, especially weak serum reactions. We evaluated the discrepant results by the reference manual method to confirm ABO blood typing. Methods In total, 13,113 blood samples were tested with the AutoVue system; all samples were run in parallel with the reference manual method according to the laboratory protocol. Results The AutoVue system confirmed ABO blood typing of 12,816 samples (97.7%), and these results were concordant with those of the manual method. The remaining 297 samples (2.3%) showed discrepant results in the AutoVue system and were confirmed by the manual method. The discrepant results involved weak serum reactions (<2+ reaction grade), extra serum reactions, samples from patients who had received stem cell transplants, ABO subgroups, and specific system error messages. Among the 98 samples showing ≤1+ reaction grade in the AutoVue system, 70 samples (71.4%) showed a normal serum reaction (≥2+ reaction grade) with the manual method, and 28 samples (28.6%) showed weak serum reaction in both methods. Conclusions ABO blood tying of 97.7% samples could be confirmed by the AutoVue system and a small proportion (2.3%) needed to be re-evaluated by the manual method. Samples with a 2+ reaction grade in serum typing do not need to be evaluated manually, while those with ≤1+ reaction grade do. PMID:28028997

  7. [Private umbilical cord blood banking does not reduce the number of samples for scientific stem cell research].

    PubMed

    Jacobs, V R; Niemeyer, M; Gottschalk, N; Schneider, K T; Kiechle, M

    2005-12-01

    Private umbilical cord blood (UCB) banking after delivery has increased over the last decade. For adult/somatic stem cell research UCB is an essential source of stem cells and researchers question if the number of UCB samples for research might be reduced by private banking. A survey among seven private blood banks in Germany and analysis and comparison of the number of UCB samples donated for research within the STEMMAT project with private blood banking were performed from 03/2003 to 06/2005 at the Frauenklinik (OB/GYN), Technical University Munich, Germany. Within 27.5 months 1,551 UCB samples were collected for research purposes; the effective recruitment rate was higher than expectations at an effective 66.2 %. Private UCB banking [n = 24] was distributed among three cord blood banks [n = 16, 6 and 4]. The rate of private blood banking was 0.99 % for all deliveries, thus reducing the effective rate for research purpose by only 1.5 %. Under the assumption of active and successful recruitment of scientific UCB samples, private blood banking does not significantly reduce this rate and therefore is a negligible rival in the competition for sufficient numbers of UCB samples for research.

  8. Utility of the microculture method for Leishmania detection in non-invasive samples obtained from a blood bank.

    PubMed

    Ates, Sezen Canim; Bagirova, Malahat; Allahverdiyev, Adil M; Kocazeybek, Bekir; Kosan, Erdogan

    2013-10-01

    In recent years, the role of donor blood has taken an important place in epidemiology of Leishmaniasis. According to the WHO, the numbers of patients considered as symptomatic are only 5-20% of individuals with asymptomatic leishmaniasis. In this study for detection of Leishmania infection in donor blood samples, 343 samples from the Capa Red Crescent Blood Center were obtained and primarily analyzed by microscopic and serological methods. Subsequently, the traditional culture (NNN), Immuno-chromatographic test (ICT) and Polymerase Chain Reaction (PCR) methods were applied to 21 samples which of them were found positive with at least one method. Buffy coat (BC) samples from 343 blood donors were analyzed: 15 (4.3%) were positive by a microculture method (MCM); and 4 (1.1%) by smear. The sera of these 343 samples included 9 (2.6%) determined positive by ELISA and 7 (2%) positive by IFAT. Thus, 21 of (6.1%) the 343 subjects studied by smear, MCM, IFAT and ELISA techniques were identified as positive for leishmaniasis at least one of the techniques and the sensitivity assessed. According to our data, the sensitivity of the methods are identified as MCM (71%), smear (19%), IFAT (33%), ELISA (42%), NNN (4%), PCR (14%) and ICT (4%). Thus, with this study for the first time, the sensitivity of a MCM was examined in blood donors by comparing MCM with the methods used in the diagnosis of leishmaniasis. As a result, MCM was found the most sensitive method for detection of Leishmania parasites in samples obtained from a blood bank. In addition, the presence of Leishmania parasites was detected in donor bloods in Istanbul, a non-endemic region of Turkey, and these results is a vital importance for the health of blood recipients.

  9. [Detection of antibodies against Trypanosoma cruzi in Somoto, Nicaragua, using indirect ELISA and IFI on blood samples on filter paper].

    PubMed

    Palacios, X; Belli, A; Espino, A M

    2000-12-01

    We standardized a solid-phase enzyme-linked immunosorbent assay (ELISA) in order to study the presence of Trypanosoma cruzi antibodies in asymptomatic persons who live in an area of Nicaragua endemic for Chagas' disease. The test was standardized to analyze filter-paper blood samples, which are easy to transport. In the first phase of our investigation, ELISA was used to study 18 samples of total serum and 18 eluates of blood from patients with chronic Chagas' disease; 30 samples of serum and 30 eluates of blood from healthy people, used as negative controls; and 14 samples of serum and 14 eluates of blood from patients with cutaneous or visceral leishmaniasis, which were used to study cross-reactions. Both with the total-serum and the blood-eluate samples, the ELISA test provided 100% sensitivity and 90% specificity. Cross-reactions in the patient samples were observed only with visceral leishmaniasis. The second phase of our investigation was a population study that included eight rural communities in the area of Somoto, Nicaragua. Through random sampling, filter-paper blood samples were collected from 2,434 people (1,335 men and 1,099 women) from the communities of Aguas Calientes, El Brocal, La Manzana, Las Playas, Los Canales, Santa Isabel, Santa Rosa, and Santa Teresa. Studied by ELISA and by indirect immunofluorescence (IIF), the samples included 260 found seropositive by ELISA (10.7%), of which 207 were positive according to IIF (8.5%). With both techniques, the majority of seropositives were among women, but the difference between men and women was not statistically significant. There was a high level of agreement between the results obtained with the two techniques. There was an upward trend with age, with 5.4% of those found seropositive by ELISA being persons 10 years of age or younger and 42.7% of those found seropositive being older than 50. The vast majority of the individuals analyzed were asymptomatic.

  10. On-chip acoustophoretic isolation of microflora including S. typhimurium from raw chicken, beef and blood samples.

    PubMed

    Ngamsom, Bongkot; Lopez-Martinez, Maria J; Raymond, Jean-Claude; Broyer, Patrick; Patel, Pradip; Pamme, Nicole

    2016-04-01

    Pathogen analysis in food samples routinely involves lengthy growth-based pre-enrichment and selective enrichment of food matrices to increase the ratio of pathogen to background flora. Similarly, for blood culture analysis, pathogens must be isolated and enriched from a large excess of blood cells to allow further analysis. Conventional techniques of centrifugation and filtration are cumbersome, suffer from low sample throughput, are not readily amenable to automation and carry a risk of damaging biological samples. We report on-chip acoustophoresis as a pre-analytical technique for the resolution of total microbial flora from food and blood samples. The resulting 'clarified' sample is expected to increase the performance of downstream systems for the specific detection of the pathogens. A microfluidic chip with three inlets, a central separation channel and three outlets was utilized. Samples were introduced through the side inlets, and buffer solution through the central inlet. Upon ultrasound actuation, large debris particles (10-100 μm) from meat samples were continuously partitioned into the central buffer channel, leaving the 'clarified' outer sample streams containing both, the pathogenic cells and the background flora (ca. 1 μm) to be collected over a 30 min operation cycle before further analysis. The system was successfully tested with Salmonella typhimurium-spiked (ca. 10(3)CFU mL(-1)) samples of chicken and minced beef, demonstrating a high level of the pathogen recovery (60-90%). When applied to S. typhimurium contaminated blood samples (10(7)CFU mL(-1)), acoustophoresis resulted in a high depletion (99.8%) of the red blood cells (RBC) which partitioned in the buffer stream, whilst sufficient numbers of the viable S. typhimurium remained in the outer channels for further analysis. These results indicate that the technology may provide a generic approach for pre-analytical sample preparation prior to integrated and automated downstream detection of

  11. Telomere length measurement by qPCR in birds is affected by storage method of blood samples.

    PubMed

    Reichert, Sophie; Froy, Hannah; Boner, Winnie; Burg, Theresa M; Daunt, Francis; Gillespie, Robert; Griffiths, Kate; Lewis, Sue; Phillips, Richard A; Nussey, Dan H; Monaghan, Pat

    2017-06-01

    Given the potential role of telomeres as biomarkers of individual health and ageing, there is an increasing interest in studying telomere dynamics in a wider range of taxa in the fields of ecology and evolutionary biology. Measuring telomere length across the lifespan in wild animal systems is essential for testing these hypotheses, and may be aided by archived blood samples collected as part of longitudinal field studies. However, sample collection, storage, and DNA extraction methods may influence telomere length measurement, and it may, therefore, be difficult to balance consistency in sampling protocol with making the most of available samples. We used two complementary approaches to examine the impacts of sample storage method on measurements of relative telomere length (RTL) by qPCR, particularly focusing on FTA (Flinders Technology Associates) cards as a long-term storage solution. We used blood samples from wandering albatrosses collected over 14 years and stored in three different ways (n = 179), and also blood samples from captive zebra finches (n = 30) that were each stored using three different methods. Sample storage method influenced RTL in both studies, and samples on FTA cards had significantly shorter RTL measurements. There was no significant correlation between RTL measured in zebra finch blood on FTA cards and the same samples stored either as frozen whole blood or as extracted DNA. These results highlight the importance of consistency of sampling protocol, particularly in the context of long-term field studies, and suggest that FTA cards should not be used as a long-term storage solution to measure RTL without validation.

  12. Detection of Theileria annulata in blood samples of native cattle by PCR and smear method in Southeast of Iran.

    PubMed

    Nourollahi-Fard, Saeid R; Khalili, Mohammad; Ghalekhani, Nima

    2015-06-01

    Theileria annulata, a protozoan parasite of cattle is causes tropical theileriosis. Polymerase chain reaction (PCR) was used to assess the presence and the frequency of T. annulata infection in blood samples obtained from carrier cattle in Kerman, Southeast of Iran. Blood samples were collected in citrate solution from 150 native cattle with mean age of 1 year which selected randomly. Primarily, a thin layer smear was prepared from their ear sublime vein blood and was fixed with methanol and stained with Giemsa dye. Blood smears were examined for the presence of parasites, and blood samples were analyzed by PCR. Piroplasmic forms of T. annulata were seen in 16 of 150 (10.66 %) by examination the blood smears with light microscope, whereas 68 of 150 (45.33 %) cattle were positive by PCR method. All animals that were positive by blood smears were also positive by PCR. Difference between these methods was significant (P < 0.05). Our results demonstrate that this PCR assay in diagnosing T. annulata parasites in carrier cattle is more sensitive than method of smear preparation and can be used in epidemiological studies.

  13. Simplified [18F]FDG Image-Derived Input Function Using the Left Ventricle, Liver, and One Venous Blood Sample

    PubMed Central

    Tantawy, Mohammed Noor; Peterson, Todd E.

    2014-01-01

    A relatively simple, almost entirely noninvasive imaging-based method is presented for deriving arterial blood input functions for quantitative [18F]2-fluoro-2-deoxy-D-glucose (FDG) positron emission tomographic (PET) studies in rodents. It requires only one venous blood sample at the end of the scan. MicroPET images and arterial blood time-activity curves (TACs) were downloaded from the Mouse Quantitation Program database at the University of California, Los Angeles. Three-dimensional regions of interest were drawn around the blood-pool region of the left ventricle and within the liver to derive their respective TACs. To construct the “hybrid” image-derived input function (IDIF), the initial part of the left ventricle TAC, containing the peak concentration of [18F]FDG in the arterial blood, was corrected for spillout (ie, partial-volume effect yielding a recovery coefficient < 1) and then joined to the liver TAC (normalized to the 60-minute arterial blood sample) immediately after it peaks. To validate our method, the [18F]FDG influx constant (Ki) was estimated using a two-tissue compartment model and compared to estimates of Ki obtained using measured arterial blood TACs. No significant difference in the Ki estimates was obtained with the arterial blood input function and our hybrid IDIF. We conclude that the normalized hybrid IDIF can be used in practice to obtain reliable Ki estimates. PMID:20236605

  14. Rapid Detection of Candida albicans by Polymerase Spiral Reaction Assay in Clinical Blood Samples

    PubMed Central

    Jiang, Xiaoqun; Dong, Derong; Bian, Lihong; Zou, Dayang; He, Xiaoming; Ao, Da; Yang, Zhan; Huang, Simo; Liu, Ningwei; Liu, Wei; Huang, Liuyu

    2016-01-01

    Candida albicans is the most common human yeast pathogen which causes mucosal infections and invasive fungal diseases. Early detection of this pathogen is needed to guide preventative and therapeutic treatment. The aim of this study was to establish a polymerase spiral reaction (PSR) assay that rapidly and accurately detects C. albicans and to assess the clinical applicability of PSR-based diagnostic testing. Internal transcribed spacer 2 (ITS2), a region between 5.8S and 28S fungal ribosomal DNA, was used as the target sequence. Four primers were designed for amplification of ITS2 with the PSR method, which was evaluated using real time turbidity monitoring and visual detection using a pH indicator. Fourteen non-C. albicans yeast strains were negative for detection, which indicated the specificity of PSR assay was 100%. A 10-fold serial dilution of C. albicans genomic DNA was subjected to PSR and conventional polimerase chain reaction (PCR) to compare their sensitivities. The detection limit of PSR was 6.9 pg/μl within 1 h, 10-fold higher than that of PCR (69.0 pg/μl). Blood samples (n = 122) were collected from intensive care unit and hematological patients with proven or suspected C. albicans infection at two hospitals in Beijing, China. Both PSR assay and the culture method were used to analyze the samples. Of the 122 clinical samples, 34 were identified as positive by PSR. The result was consistent with those obtained by the culture method. In conclusion, a novel and effective C. albicans detection assay was developed that has a great potential for clinical screening and point-of-care testing. PMID:27379048

  15. Pharmacokinetics and pharmacodynamics of recombinant human erythropoietin in athletes. Blood sampling and doping control

    PubMed Central

    SOUILLARD, AGNES; AUDRAN, MICHEL; BRESSOLLE, FRANÇOISE; GAREAU, RAYNALD; DUVALLET, ALAIN; CHANAL, JEAN-LOUIS

    1996-01-01

    1The pharmacokinetics of recombinant human erythropoietin (rHuEpo) were initially determined in two healthy volunteers after a single subcutaneous dose (50 u kg−1). Twenty subjects then received repeated subcutaneous administrations of high dose (200 u kg−1) rHuEpo and 10 subjects received placebo. An immunoradiometric assay was used to measure the concentrations of erythropoietin (Epo) in serum and urine. 2Serum Epo concentration-time profiles were best described by a one-compartment open model with zero-order input. The mean elimination half-life (±s.d.) was 42.0±34.2 h. Clearance, uncorrected for bioavailability, was 0.05±0.01 l h−1 kg& minus;1. Erythropoietin concentrations returned to normal values in serum and urine, 7 and 4 days after the last administration, respectively. 3The recombinant hormone was well tolerated. Significant changes in reticulocytes and red blood cells, haemoglobin concentrations and haematocrit were observed after administration of rHuEpo. In the control group, these parameters remained unchanged. 4The change in reticulocytes was used as an index of the therapeutic effect of rHuEpo. The concentration-effect relationship was best described by an exponential model. 5These data show the limitations of the measurement of Epo concentrations in blood and urine samples, collected in athletes during competition, for antidoping control. Epo doping can be detected only during or within 4 to 7 days of ending, a course of rHuEpo. PMID:8877027

  16. Coagulation indices in very preterm infants from cord blood and postnatal samples.

    PubMed

    Neary, E; McCallion, N; Kevane, B; Cotter, M; Egan, K; Regan, I; Kirkham, C; Mooney, C; Coulter-Smith, S; Ní Áinle, F

    2015-11-01

    Very premature infants are at high risk of bleeding complications; however, few data exist on ranges for standard coagulation tests. The primary objective of this study was to measure standard plasma coagulation tests and thrombin generation in very premature infants compared with term infants. The secondary objective was to evaluate whether an association existed between coagulation indices and intraventricular hemorrhage (IVH). Cord and peripheral blood of neonates < 30 weeks gestational age (GA) was drawn at birth, on days 1 and 3 and fortnightly until 30 weeks corrected gestational age. Prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen and coagulation factor levels were measured and tissue factor-stimulated thrombin generation was characterized. Control plasma was obtained from cord blood of term neonates. One hundred and sixteen infants were recruited. Median (range) GA was 27.7 (23.7-29.9) weeks and mean (SD) birth weight was 1020 (255) g. Median (5th-95th percentile) day 1 PT, APTT and fibrinogen were 17.5 (12.7-26.6) s, 78.7 (48.7-134.3) s and 1.4 (0.72-3.8) g L(-1) , respectively. No difference in endogenous thrombin potential between preterm and term plasma was observed, where samples were available. Levels of coagulation factors II, VII, IX and X, protein C, protein S and antithrombin were reduced in preterm compared with term plasma. Day 1 APTT and PT were not associated with IVH. In the largest cross-sectional study to date of very preterm infants, typical ranges for standard coagulation tests were determined. Despite long clotting times, thrombin generation was observed to be similar in very preterm and term infants. © 2015 International Society on Thrombosis and Haemostasis.

  17. Parametric images of antibody pharmacokinetics based on serial quantitative whole-body imaging and blood sampling.

    PubMed

    Gleisner, Katarina Sjögreen; Nickel, Mattias; Lindén, Ola; Erlandsson, Kjell; Wingårdh, Karin; Strand, Sven-Erik

    2007-08-01

    We present a method for pharmacokinetic modeling of distributions of (111)In-labeled monoclonal antibodies (mAbs) on individual pixels of planar scintillation-camera images. The method is applied to 2 sets of clinical whole-body images, each consisting of 6 consecutive images acquired over a week. Quantification is performed on a pixel basis, yielding images in units of Bq/pixel. The images acquired on the different occasions are registered using a nonrigid method, and for each pixel location a time-activity curve is obtained for which kinetic modeling is performed. The (111)In-mAb is assumed to be located in either the vascular or the extravascular space. The vascular content is assumed to follow the global blood kinetics as determined from blood samples, together with a model parameter alpha that describes the fraction of the whole-body blood volume present in the particular pixel. The rate of change of the extravascular compartment is described by a linear 1-tissue-compartment model with 2 rate constants, K'1 and k2, reflecting extravasation and washout, respectively. The model is optimized for each pixel position with regard to the values of the 3 parameters (alpha, K'1, and k2), resulting in 3 parametric images. From these, images of the cumulated activity in vascular and extravascular spaces are calculated, as is an image of the rate-constants ratio, which is closely related to the volume of distribution. The resulting parametric images are analyzed in terms of the appearance of the time-activity curves at various locations. Results also include interpretation of the parametric images in their clinical context, and the location of regions that exhibit high extravasation and a low washout rate is compared with confirmed malignant sites. Parametric imaging allows the study and analysis of the spatial and temporal distributions of mAbs simultaneously. Parametric imaging enhances regions where the pharmacokinetics differ from the surrounding tissue and provides a

  18. Associations between Dietary Patterns and Blood Pressure in a Clinical Sample of Overweight Adults.

    PubMed

    Ndanuko, Rhoda N; Tapsell, Linda C; Charlton, Karen E; Neale, Elizabeth P; Batterham, Marijka J

    2017-02-01

    Dietary pattern analysis provides important evidence revealing diet-disease relationships. It may be especially useful in areas less well researched, such as diet and hypertension in clinical populations. The aim of this study was to identify the association between dietary patterns and blood pressure (BP) in a sample of overweight adults volunteering for a clinical trial for weight loss. This cross-sectional analysis used baseline data from the HealthTrack study, a 12-month randomized controlled trial. Dietary intake was evaluated with 4-day food records. Participants were 328 adults recruited from the Illawarra region of New South Wales, Australia, between May 2014 and April 2015. Resting BP and 24-hour urine sodium and potassium were measured. Dietary patterns were derived by principal component analysis from 21 food groups. Multiple regression analysis was performed to assess the association between the extracted dietary patterns and BP. The participants' mean age was 43.6±8.0 years, mean body mass index was 32.4±4.2, and mean systolic BP/diastolic BP was 124.9±14.5/73.3±9.9 mm Hg. Six major dietary patterns were identified: "nuts, seeds, fruit, and fish," "milk and meat," "breads, cereals, and snacks," "cereal-based products, fats, and oils," "alcohol, eggs, and legumes," and "savoury sauces, condiments, and meat." The "nuts, seeds, fruit, and fish" dietary pattern was significantly and inversely associated with systolic BP (F [7,320]=15.248; P<0.0005; adjusted R(2)=0.234 and diastolic BP (F [7,320]=17.351; P<0.0005; adjusted R(2)=0.259) and sodium-to-potassium ratio (F [7,320]=6.210; P<0.0005; adjusted R(2)=0.100). A dietary pattern rich in nuts, seeds, fruit, and fish was inversely associated with blood pressure in this clinical sample. The findings suggest that current dietary guidelines are relevant to an overweight clinical population and support the value of dietary pattern analysis when exploring the diet-disease relationship. Copyright © 2017

  19. PCR-Based Detection of Toxoplasma gondii DNA in Blood and Ocular Samples for Diagnosis of Ocular Toxoplasmosis

    PubMed Central

    Bourdin, C.; Busse, A.; Kouamou, E.; Touafek, F.; Bodaghi, B.; Le Hoang, P.; Mazier, D.; Paris, L.

    2014-01-01

    PCR detection of Toxoplasma gondii in blood has been suggested as a possibly efficient method for the diagnosis of ocular toxoplasmosis (OT) and furthermore for genotyping the strain involved in the disease. To assess this hypothesis, we performed PCR with 121 peripheral blood samples from 104 patients showing clinical and/or biological evidence of ocular toxoplasmosis and from 284 (258 patients) controls. We tested 2 different extraction protocols, using either 200 μl (small volume) or 2 ml (large volume) of whole blood. Sensitivity was poor, i.e., 4.1% and 25% for the small- and large-volume extractions, respectively. In comparison, PCR with ocular samples yielded 35.9% sensitivity, while immunoblotting and calculation of the Goldmann-Witmer coefficient yielded 47.6% and 72.3% sensitivities, respectively. Performing these three methods together provided 89.4% sensitivity. Whatever the origin of the sample (ocular or blood), PCR provided higher sensitivity for immunocompromised patients than for their immunocompetent counterparts. Consequently, PCR detection of Toxoplasma gondii in blood samples cannot currently be considered a sufficient tool for the diagnosis of OT, and ocular sampling remains necessary for the biological diagnosis of OT. PMID:25210066

  20. Human Milk Oligosaccharides and Lewis Blood Group: Individual High-Throughput Sample Profiling to Enhance Conclusions From Functional Studies12

    PubMed Central

    Blank, Dennis; Dotz, Viktoria; Geyer, Rudolf; Kunz, Clemens

    2012-01-01

    Human milk oligosaccharides (HMO) are discussed to play a crucial role in an infant’s development. Lewis blood group epitopes, in particular, seem to remarkably contribute to the beneficial effects of HMO. In this regard, large-scale functional human studies could provide evidence of the variety of results from in vitro investigations, although increasing the amount and complexity of sample and data handling. Therefore, reliable screening approaches are needed. To predict the oligosaccharide pattern in milk, the routine serological Lewis blood group typing of blood samples can be applied due to the close relationship between the biosynthesis of HMO and the Lewis antigens on erythrocytes. However, the actual HMO profile of the individual samples does not necessarily correspond to the serological determinations. This review demonstrates the capabilities of merging the traditional serological Lewis blood group typing with the additional information provided by the comprehensive elucidation of individual HMO patterns by means of state-of-the-art analytics. Deduced from the association of the suggested HMO biosynthesis with the Lewis blood group, the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry profiles of oligosaccharides in individual milk samples exemplify the advantages and the limitations of sample assignment to distinct groups. PMID:22585923

  1. Hemoglobin Variant Analysis via Direct Surface Sampling of Dried Blood Spots Coupled with High-Resolution Mass Spectrometry

    PubMed Central

    2011-01-01

    Hemoglobinopathies are the most common inherited disorders. Newborn blood screening for clinically significant hemoglobin variants, including sickle (HbS), HbC, and HbD, has been adopted in many countries as it is widely acknowledged that early detection improves the outcome. We present a method for determination of Hb variants by direct surface sampling of dried blood spots by use of an Advion Triversa Nanomate automated electrospray system coupled to a high-resolution mass spectrometer. The method involves no sample preparation. It is possible to unambiguously identify homozygous and heterozygous HbS, HbC, and HbD variants in <10 min without the need for additional confirmation. The method allows for repeated analysis of a single blood spot over a prolonged time period and is tolerant of blood spot storage conditions. PMID:21341716

  2. [Control of veterinary communicable diseases in swine and fattening cattle stocks by serological testing of slaughtered animal blood samples].

    PubMed

    Pannwitz, S; Riechert, D; Wolter, F; Franze, F

    1981-01-01

    Economy of prophylactic haemoserological control of pig stocks in a district as well as of a sow breeding unit and of cattle stocks on two industrialised fattening farms has been enhanced by including blood samples from routine slaughtering. Preliminary organisational experience is reported in this paper. The samples collected samples collected from normally slaughtered selected sows may contribute to an improvement of veterinary production control.

  3. Blood and urine samples as useful sources for the direct detection of tuberculosis by polymerase chain reaction.

    PubMed

    Rebollo, María J; San Juan Garrido, Rafael; Folgueira, Dolores; Palenque, Elia; Díaz-Pedroche, C; Lumbreras, Carlos; Aguado, José M

    2006-10-01

    The aim of the study was to assess the utility of the polymerase chain reaction (PCR) assay in blood and urine for the diagnosis of tuberculosis (TB). We prospectively evaluated the usefulness of PCR performed in blood and urine samples from patients with proved or probable TB compared with a control group of patients. The PCR technique was performed using IS6110 primers. We included in the study 57 patients (43 with definite TB and 14 with probable TB) and 26 controls. Blood and urine samples were drawn at the time of microbiologic diagnosis and 3, 6, 9, and 12 months later. Cultures were positive in the early period (<1 month after treatment) in 11 of 57 patients (19%) with probable or definite TB, in comparison with 42% of patients (24/57) who yielded a positive PCR (P = 0.02). Urine samples increased the sensitivity of PCR determination in blood samples by 10%. The PCR in blood and/or urine was positive in 41% of patients with pulmonary TB, in 36% of patients with extrapulmonary TB, and in 50% of patients with disseminated TB. Mycobacterium tuberculosis was still detectable by PCR in 5 of 13 patients with cured TB after 1 or more months of antituberculous treatment. The PCR detection of M. tuberculosis in blood and urine samples is useful for the diagnosis of different clinical forms of TB, mostly in those patients in which sample extraction is difficult or requires aggressive techniques. The sensitivity of this technique could be improved studying more than 1 sample in each patient, even after initiating an antituberculous treatment.

  4. Effect of shear stress in the flow through the sampling needle on concentration of nanovesicles isolated from blood.

    PubMed

    Štukelj, Roman; Schara, Karin; Bedina-Zavec, Apolonija; Šuštar, Vid; Pajnič, Manca; Pađen, Ljubiša; Krek, Judita Lea; Kralj-Iglič, Veronika; Mrvar-Brečko, Anita; Janša, Rado

    2017-02-15

    During harvesting of nanovesicles (NVs) from blood, blood cells and other particles in blood are exposed to mechanical forces which may cause activation of platelets, changes of membrane properties, cell deformation and shedding of membrane fragments. We report on the effect of shear forces imposed upon blood samples during the harvesting process, on the concentration of membrane nanovesicles in isolates from blood. Mathematical models of blood flow through the needle during sampling with vacuumtubes and with free flow were constructed, starting from the Navier-Stokes formalism. Blood was modeled as a Newtonian fluid. Work of the shear stress was calculated. In experiments, nanovesicles were isolated by repeated centrifugation (up to 17,570×g) and washing, and counted by flow cytometry. It was found that the concentration of nanovesicles in the isolates positively corresponded with the work by the shear forces in the flow of the sample through the needle. We have enhanced the effect of the shear forces by shaking the samples prior to isolation with glass beads. Imaging of isolates by scanning electron microscopy revealed closed globular structures of a similar size and shape as those obtained from unshaken plasma by repetitive centrifugation and washing. Furthermore, the sizes and shapes of NVs obtained by shaking erythrocytes corresponded to those isolated from shaken platelet-rich plasma and from unshaken platelet rich plasma, and not to those induced in erythrocytes by exogenously added amphiphiles. These results are in favor of the hypothesis that a significant pool of nanovesicles in blood isolates is created during their harvesting. The identity, shape, size and composition of NVs in isolates strongly depend on the technology of their harvesting. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Establishment of experimental conditions for preserving samples of fish blood for analysis with both comet assay and flow cytometry.

    PubMed

    Ramsdorf, Wanessa A; Guimarães, Fernando de S F; Ferraro, Marcos V M; Gabardo, Juarez; Trindade, Edvaldo da Silva; Cestari, Marta Margarete

    2009-02-19

    When environmental analysis is performed, the high number of samples required and handling conditions during the transport of these samples to the laboratory are common problems. The comet assay is a useful, highly sensitive tool in biomonitoring. Some studies in the literature aim to preserve slides in lysis solution for use in the comet assay. Until now, however, no efficient methodology for preserving blood samples for this assay has been described. Because of this, the present report aimed to establish the proper conditions for samples maintenance prior to comet assay analysis. Samples were conserved in three different solutions: a high protein concentration solution (fetal bovine serum-FBS), an anticoagulant agent (a calcium chelator - ethylenediaminetetracetic acid - EDTA), and a salt buffered solution (phosphate buffered saline-PBS). Therefore, peripheral blood samples of Rhamdia quelen specimens were collected and maintained in these solutions until testing at 72h. Analyses of DNA fragmentation via the comet assay and cell viability via flow cytometry were performed at intervals of 24h. The results showed that samples maintained in FBS were preserved better; this was followed by those preserved in PBS and then last by those preserved in EDTA. In conclusion, blood samples from freshwater fish can be preserved up to 48h in fetal bovine serum at 4 degrees C in the absence of light. In this period, no DNA fragmentation occurs. We thus describe an excellent method of sample conservation for subsequent analysis in the laboratory.

  6. Validation of a fully automated robotic setup for preparation of whole blood samples for LC-MS toxicology analysis.

    PubMed

    Andersen, David; Rasmussen, Brian; Linnet, Kristian

    2012-05-01

    A fully automated setup was developed for preparing whole blood samples using a Tecan Evo workstation. By integrating several add-ons to the robotic platform, the flexible setup was able to prepare samples from sample tubes to a 96-well sample plate ready for injection on liquid chromatography-mass spectrometry using several preparation techniques, including protein precipitation, solid-phase extraction and centrifugation, without any manual intervention. Pipetting of a known aliquot of whole blood was achieved by integrating a balance and performing gravimetric measurements. The system was able to handle 1,073 of 1,092 (98.3%) samples of whole blood from forensic material, including postmortem samples, without any need for repeating sample preparation. Only three samples required special treatment such as dilution. The addition of internal and calibration standards were validated by pipetting a solution of Orange G and measuring the weight and absorbance. Internal standard (20 µL) was added in a multi-pipetting sequence with an accuracy of 99.9% and imprecision (coefficient of variation) of 1.6%. Calibration standards were added with high accuracy at volumes as low as 6.00 µL (±0.21 µL). The general setup of the offline sample preparation and key validation parameters of a quantitative analysis of Δ(9)-tetrahydrocannabinol is presented.

  7. A dried blood spots technique based LC-MS/MS method for the analysis of posaconazole in human whole blood samples.

    PubMed

    Reddy, Todime M; Tama, Cristina I; Hayes, Roger N

    2011-11-15

    A rugged and robust liquid chromatographic tandem mass spectrometric (LC-MS/MS) method utilizing dried blood spots (DBS) was developed and validated for the analysis of posaconazole in human whole blood. Posaconazole fortified blood samples were spotted (15 μL) onto Ahlstrom Alh-226 DBS cards and dried for at least 2h. Punched spots were then extracted by using a mixture of acetonitrile and water containing stable labeled internal standard (IS). Posaconazole and its IS were separated from endogenous matrix components on a Kinetex™ C18 column under gradient conditions with a mobile phase A consisting of 0.1% formic acid and a mobile phase B consisting of 0.1% formic acid in acetonitrile/methanol (70/30, v/v). The analyte and IS were detected using a Sciex API 4000 triple quadrupole LC-MS/MS system equipped with a TurboIonSpray™ source operated in the positive ion mode. The assay was linear over the concentration range of 5-5000 ng/mL. The inter-run accuracy and precision of the assay were -1.8% to 0.8% and 4.0% to 10.4%, respectively. Additional assessments unique to DBS were investigated including sample spot homogeneity, spot volume, and hematocrit. Blood spot homogeneity was maintained and accurate and precise quantitation results were obtained when using a blood spot volume of between 15 and 35 μL. Human blood samples with hematocrit values ranging between 25% and 41% gave acceptable quantitation results. The validation results indicate that the method is accurate, precise, sensitive, selective and reproducible.

  8. Validation of a minimally invasive blood-sampling technique for the analysis of hormones in domestic rabbits, Oryctolagus cuniculus (Lagomorpha).

    PubMed

    Voigt, Christian C; Fassbender, Mirja; Dehnhard, Martin; Wibbelt, Gudrun; Jewgenow, Katarina; Hofer, Heribert; Schaub, Günter A

    2004-01-01

    Previous studies in small mammals showed that blood-sucking bugs (Reduviidae, Heteroptera) can be used to obtain blood from veins difficult to access by human experimenters. In the present study, we validated the use of reduviid bugs for endocrinological studies in endotherms using domestic rabbits as a model organism. Two processes could alter the hormone concentrations in the blood ingested by the bug: (1) Mixing of ingested blood with saliva, gut fluid, or hemolymph and (2) digestive processes. We compared concentrations of progesterone, testosterone, and hydrocortisone in blood samples that were acquired from domestic rabbits (Oryctolagus cuniculus) by bugs (Dipetalogaster maxima) with hormone concentrations in blood obtained from the same individual rabbits with a conventional method, i.e., syringe. We found no significant differences in hormone concentrations between the two methods. Thus, the mixing effect is negligible immediately after the blood meal. In addition, we also could not find significant changes in concentrations of progesterone and hydrocortisone for up to 8h after the blood meal. Whereas levels of hydrocortisone remained unchanged for even 24h, progesterone levels significantly increased between eight and 24h. Thus, the bugs' excretory apparatus did not fractionate between water and hormones. Thirdly, we hypothesized that reduviid bugs impose less stress on the rabbits than the conventional method. We showed that deviations in hydrocortisone concentrations between the two blood sampling routines were lower when the bug method was used first and higher when the conventional method was used first. Thus, bugs imposed less stress on the study animals than the conventional method. Overall, we conclude that reduviid bugs present a minimally invasive method for obtaining blood from endotherm animals for endocrinological studies.

  9. Adaptive sampling of CT data for myocardial blood flow estimation from dose-reduced dynamic CT

    NASA Astrophysics Data System (ADS)

    Modgil, Dimple; Bindschadler, Michael D.; Alessio, Adam M.; La Rivière, Patrick J.

    2015-03-01

    Quantification of myocardial blood flow (MBF) can aid in the diagnosis and treatment of coronary artery disease (CAD). However, there are no widely accepted clinical methods for estimating MBF. Dynamic CT holds the promise of providing a quick and easy method to measure MBF quantitatively, however the need for repeated scans has raised concerns about the potential for high radiation dose. In our previous work, we explored techniques to reduce the patient dose by either uniformly reducing the tube current or by uniformly reducing the number of temporal frames in the dynamic CT sequence. These dose reduction techniques result in very noisy data, which can give rise to large errors in MBF estimation. In this work, we seek to investigate whether nonuniformly varying the tube current or sampling intervals can yield more accurate MBF estimates. Specifically, we try to minimize the dose and obtain the most accurate MBF estimate through addressing the following questions: when in the time attenuation curve (TAC) should the CT data be collected and at what tube current(s). We hypothesize that increasing the sampling rate and/or tube current during the time frames when the myocardial CT number is most sensitive to the flow rate, while reducing them elsewhere, can achieve better estimation accuracy for the same dose. We perform simulations of contrast agent kinetics and CT acquisitions to evaluate the relative MBF estimation performance of several clinically viable adaptive acquisition methods. We found that adaptive temporal and tube current sequences can be performed that impart an effective dose of about 5 mSv and allow for reductions in MBF estimation RMSE on the order of 11% compared to uniform acquisition sequences with comparable or higher radiation doses.

  10. Trace samples of human blood in mosquitoes as a forensic investigation tool.

    PubMed

    Rabêlo, K C N; Albuquerque, C M R; Tavares, V B; Santos, S M; Souza, C A; Oliveira, T C; Oliveira, N C L; Crovella, S

    2015-11-23

    Investigations of any type of crime invariably starts at the crime scene by collecting evidence. Thus, the purpose of this research was to collect and analyze an entomological trace from an environment that is similar to those of indoor crime scenes. Hematophagous mosquitoes were collected from two residential units; saliva of volunteers that were residents in the units was also collected for genetic analysis as reference samples. We examined the allele frequencies of 15 short tandem repeat loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA) and amelogenin. A total of 26 female hematophagous mosquitoes were identified as Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus; we were able to obtain 11 forensically valid genetic profiles, with a minimum of 0.028203 ng/μL of human DNA. Thus, the results of this study showed that it was possible to correlate human genetic information from mosquitoes with the volunteer reference samples, which validates the use of this information as forensic evidence. Furthermore, we observed mixed genetic profiles from one mosquito. Therefore, it is clearly important to collect these insects indoors where crimes were committed, because it may be possible to find intact genetic profiles of suspects in the blood found in the digestive tract of hematophagous mosquitoes for later comparison to identify an offender and/or exclude suspects.

  11. Selective nonenzymatic bilirubin detection in blood samples using a Nafion/Mn-Cu sensor.

    PubMed

    Noh, Hui-Bog; Won, Mi-Sook; Shim, Yoon-Bo

    2014-11-15

    The specific detection of biological organics without the use of an enzyme is challenging, and it is crucial for analytical and clinical chemistry. We report specific nonenzymatic bilirubin detection through the catalytic oxidation of bilirubin molecule on the Nafion/Mn-Cu surface. The catalytic ability, true surface area, morphology, crystallinity, composition, and oxidation state of the sensor surface were assessed using voltammetry, coulometry, XPS, XRD, Brunauer-Emmett-Teller (BET), SEM, EDXS, and TOF-SIMS experiments. The results showed that the surface was composed of microporous Mn-Cu bimetallic crystal in flake shape with a large BET surface area (3.635 m(2)g(-1)), where the surface area and crystallinity mainly affected the sensor performance. Product analysis of the catalytic reaction on the sensor probe revealed a specific two-electron oxidation of dipyrromethane moiety to dipyrromethene in the bilirubin molecule. Experimental variables affecting the analysis of bilirubin were optimized in terms of probe composition, temperature, pH, and potential. At the optimized condition, the dynamic range was between 1.2 μM and 0.42 mM, which yielded the equation of ΔI (μA)=(1.03 ± 0.72)+(457.0 ± 4.03) [C] (mM) with 0.999 of correlation coefficient, and the detection limit was 25.0 ± 1.8 nM (n=5, k=3). The stability test, interference effects, and analysis of real clinical samples, human whole blood and certified serum samples were demonstrated to confirm the reliability of the proposed bilirubin sensor.

  12. The comparison of heparinized insulin syringes and safety-engineered blood gas syringes used in arterial blood gas sampling in the ED setting (randomized controlled study).

    PubMed

    Baskın, Sevcan Baki; Oray, Neşe Çolak; Yanturalı, Sedat; Bayram, Başak

    2014-05-01

    The arterial blood gas measurement process is a painful and invasive procedure, often uncomfortable for both the patient and the physician. Because the patient-related factors that determine the difficulty of the process cannot be controlled, the physician-related factors and blood gas measurement techniques are a modifiable area of improvement that ought to be considered. Many hospitals use insulin syringes or syringes washed with heparin for the purpose of blood gas measurement because they do not have blood gas-specific syringes. In this prospective cross-sectional study, we aimed to compare safety-engineered blood gas syringes and conventional heparinized syringes used during the arterial blood gas extraction process in terms of ease of operation, the physician-patient satisfaction, laboratory appropriateness, and complications. Our study included patients whose arterial blood gas needed to be measured in the emergency department and who agreed to participate in the study. Patients were randomly divided into 2 groups. The arterial blood gas of the patients from the first group was measured by using conventional heparinized syringes, whereas safety-engineered blood gas syringes were used to measure the arterial blood gas of the patients from the second group. The groups were compared in terms of demographic data, the number of attempts, the physician and patient satisfaction, early and late-term complications, and laboratory appropriateness of the taken sample. A total of 550 patients were included in our study in a 2-month study period. There were no significant differences between patients in terms of sex, age, weight, height, body mass index, and wrist circumference. In addition, the number of attempts (P=.489), patients' pain level during the procedure (P=.145), and the degree of difficulty of the procedure according to the patient (P=.109) and physician (P=.554) were not significantly different between the groups. After arterial blood gas extraction

  13. Development of a statistical model for predicting the ethanol content of blood from measurements on saliva or breath samples.

    PubMed

    Ruz, J; Linares, P; Luque de Castro, M D; Caridad, J M; Valcarcel, M

    1989-01-01

    Blood, saliva and breath samples from a population of males and females subjected to the intake of preselected amounts of ethanol, whilst in different physical conditions (at rest, after physical exertion, on an empty stomach and after eating), were analysed by automatic methods employing immobilized (blood) or dissolved (saliva) enzymes and a breathanalyser. Treatment of the results obtained enabled the development of a statistical model for prediction of the ethanol concentration in blood at a given time from the ethanol concentration in saliva or breath obtained at a later time.

  14. Detection of sodium and potassium in single human red blood cells by 193-nm laser ablative sampling: a feasibility demonstration.

    PubMed

    Ng, C W; Cheung, N H

    2000-01-01

    The feasibility of quantifying sodium and potassium in single human erythrocytes was demonstrated by spectrochemical analysis of emissions from plasmas produced by 193-nm laser ablation of blood cells confined in a sheath flow. In one scheme, single blood cells that happened to be in the ablation volume were sampled. In another scheme, individual blood cells were first sighted and then synchronously ablated downstream. Plasma emission spectra of single ablated cells were captured, and the ratios of the analyte line intensity to the root-mean-square fluctuation of the continuum background were measured to be about 18 for sodium and 30 for potassium.

  15. Detection of canine distemper virus in blood samples by reverse transcription loop-mediated isothermal amplification.

    PubMed

    Cho, H S; Park, N Y

    2005-11-01

    Reverse transcription loop-mediated isothermal amplification (RT-LAMP) was used to detect canine distemper virus (CDV) genomic RNA. A set of four primers, two outer and two inner, were designed from CDV genomic RNA targeting the nucleocapsid protein gene. The optimal reaction time and temperature for LAMP were determined to be 60 min at 65 degrees C. The relative sensitivity and specificity of RT-LAMP was found to be 100% and 93.3%, respectively, based on 50 canine blood samples and using RT-PCR as the gold standard. The detection limit of the RT-LAMP method was 100