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Sample records for blood spot samples

  1. Performance of an app measuring spot quality in dried blood spot sampling

    NARCIS (Netherlands)

    Veenhof, Herman

    2016-01-01

    Introduction: The Dried Blood Spot sampling (DBS) method gives patients and health care workers the opportunity for remote sampling using a drop of blood from a fingerprick on a sampling card which can be send to the laboratory by mail. Laboratory analysts frequently reject DBS samples because of

  2. Quantification of multiple elements in dried blood spot samples

    DEFF Research Database (Denmark)

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

    2017-01-01

    in venous blood. Samples with different hematocrit were spotted onto filter paper to assess hematocrit effect. RESULTS: 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......BACKGROUND: 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. METHODS: 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...

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

    Science.gov (United States)

    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.

  4. Dried blood spots of pooled samples for RHD gene screening in blood donors of mixed ancestry.

    Science.gov (United States)

    Silva-Malta, M C F; Araujo, N C Fidélis; Vieira, O V Neves; Schmidt, L Cayres; Gonçalves, P de Cassia; Martins, M Lobato

    2015-10-01

    In this study, we present a strategy for RHD gene screening based on real-time polymerase chain reaction (PCR) using dried blood spots of pooled samples. Molecular analysis of blood donors may be used to detect RHD variants among the presumed D-negative individuals. RHD genotyping using pooled samples is a strategy to test a large number of samples at a more reasonable cost. RHD gene detection based on real-time PCR using dried blood spots of pooled samples was standardised and used to evaluate 1550 Brazilian blood donors phenotyped as RhD-negative. Positive results were re-evaluated by retesting single samples using real-time PCR and conventional multiplex PCR to amplify five RHD-specific exons. PCR-sequence-specific primers was used to amplify RHDψ allele. We devised a strategy for RHD gene screening using dried blood spots of five pooled samples. Among 1550 serologically D-negative blood donors, 58 (3.74%) had the RHD gene. The non-functional RHDψ allele was detected in 47 samples (3.02%). The present method is a promising strategy to detect the RHD gene among presumed RhD-negative blood donors, particularly for populations with African ancestry. © 2015 British Blood Transfusion Society.

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

    Directory of Open Access Journals (Sweden)

    Stuart A Batterman

    2016-04-01

    Full Text Available 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 (R2 > 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

  6. Dried blood spots on carboxymethyl cellulose sheets: Rapid sample preparation based on dissolution and precipitation

    DEFF Research Database (Denmark)

    Skoglund Ask, Kristine; Pedersen-Bjergaard, Stig; Gjelstad, Astrid

    2016-01-01

    This short communication describes the use of carboxymethyl cellulose sheets as sampling material for dried blood spots. Whole blood, spiked with quetiapine, a hydrophobic and basic small molecule drug substance, was spotted on the sheet and subsequently dried. The dried spot was then almost...... completely dissolved in acidified aqueous solution. It was shown that the dissolved polymer, together with major blood components can easily be precipitated and removed with acetonitrile. The presented sampling on a water-soluble biopolymer derivative followed by precipitation resulted in a simple protocol...

  7. Effects of blood sample handling procedures on measurable inflammatory markers in plasma, serum and dried blood spot samples

    DEFF Research Database (Denmark)

    Skogstrand, K.; Thorsen, P.; Vogel, I.

    2008-01-01

    increased when blood samples were stored for a period of time before the centrifugation, for certain cytokines more than 1000 fold compared to serum and plasma isolated and frozen immediately after venepuncture. The concentrations in serum generally increased more than in plasma. The measurable......The interests in monitoring inflammation by immunoassay determination of blood inflammatory markers call for information on the stability of these markers in relation to the handling of blood samples. The increasing use of stored biobank samples for such ventures that may have been collected...... and stored for other purposes, justifies the study hereof. Blood samples were stored for 0, 4, 24, and 48 h at 4 degrees C, room temperature (RT), and at 35 degrees C, respectively, before they were separated into serum or plasma and frozen. Dried blood spot samples (DBSS) were stored for 0, 1, 2, 3, 7...

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

    Science.gov (United States)

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

    2010-07-01

    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.

  9. Plasmodium falciparum HRP2 ELISA for analysis of dried blood spot samples in rural Zambia.

    Science.gov (United States)

    Gibson, Lauren E; Markwalter, Christine F; Kimmel, Danielle W; Mudenda, Lwiindi; Mbambara, Saidon; Thuma, Philip E; Wright, David W

    2017-08-23

    Dried blood spots are commonly used for sample collection in clinical and non-clinical settings. This method is simple, and biomolecules in the samples remain stable for months at room temperature. In the field, blood samples for the study and diagnosis of malaria are often collected on dried blood spot cards, so development of a biomarker extraction and analysis method is needed. A simple extraction procedure for the malarial biomarker Plasmodium falciparum histidine-rich protein 2 (HRP2) from dried blood spots was optimized to achieve maximum extraction efficiency. This method was used to assess the stability of HRP2 in dried blood spots. Furthermore, 328 patient samples made available from rural Zambia were analysed for HRP2 using the developed method. These samples were collected at the initial administration of artemisinin-based combination therapy and at several points following treatment. An average extraction efficiency of 70% HRP2 with a low picomolar detection limit was achieved. In specific storage conditions HRP2 was found to be stable in dried blood spots for at least 6 months. Analysis of patient samples showed the method to have a sensitivity of 94% and a specificity of 89% when compared with microscopy, and trends in HRP2 clearance after treatment were observed. The dried blood spot ELISA for HRP2 was found to be sensitive, specific and accurate. The method was effectively used to assess biomarker clearance characteristics in patient samples, which prove it to be ideal for gaining further insight into the disease and epidemiological applications.

  10. Genome-wide scans using archived neonatal dried blood spot samples

    Directory of Open Access Journals (Sweden)

    Wiuf Carsten

    2009-07-01

    Full Text Available Abstract Background Identification of disease susceptible genes requires access to DNA from numerous well-characterised subjects. Archived residual dried blood spot samples from national newborn screening programs may provide DNA from entire populations and medical registries the corresponding clinical information. The amount of DNA available in these samples is however rarely sufficient for reliable genome-wide scans, and whole-genome amplification may thus be necessary. This study assess the quality of DNA obtained from different amplification protocols by evaluating fidelity and robustness of the genotyping of 610,000 single nucleotide polymorphisms, using the Illumina Infinium HD Human610-Quad BeadChip. Whole-genome amplified DNA from 24 neonatal dried blood spot samples stored between 15 to 25 years was tested, and high-quality genomic DNA from 8 of the same individuals was used as reference. Results Using 3.2 mm disks from dried blood spot samples the optimal DNA-extraction and amplification protocol resulted in call-rates between 99.15% – 99.73% (mean 99.56%, N = 16, and conflicts with reference DNA in only three per 10,000 genotype calls. Conclusion Whole-genome amplified DNA from archived neonatal dried blood spot samples can be used for reliable genome-wide scans and is a cost-efficient alternative to collecting new samples.

  11. Investigations into the environmental conditions experienced during ambient sample transport: impact to dried blood spot sample shipments.

    Science.gov (United States)

    Bowen, Chester L; Dopson, Wesley; Kemp, Daniel C; Lewis, Mark; Lad, Rakesh; Overvold, Carol

    2011-07-01

    Prior to bioanalysis, sample transport and storage are critical considerations in any pharmacokinetic or toxicokinetic study design. Care must be taken to ensure the shipment is properly packaged and tracked to make certain it arrives at the desired, final destination in the appropriate timeframe, and that the integrity of the sample is not compromised. When dealing with biological specimens, environmental conditions may have a deleterious effect on the stability and conditions of the sample. Currently, frozen plasma or blood samples are the matrix of choice within the pharmaceutical industry for analysis within both preclinical and clinical trials. Liquid samples are shipped and received frozen and, therefore, the assumption is made that the frozen conditions are maintained throughout the entire transit process. Dried blood spot and dried matrix spot samples are becoming popular alternatives to plasma sampling in many small- and even large-molecule applications. With the implementation of dried blood spot and dried matrix spot samples, shipping and storage occurs under ambient conditions. In this article we discuss various shipping containers for these samples, illustrate the environmental extremes encountered during the shipping process, demonstrate a cost-effective method of monitoring both temperature and humidity, and discuss validation steps that may be implemented to minimize the impact of these variables on your study design.

  12. Protein expression profiling by antibody array analysis with use of dried blood spot samples on filter paper.

    Science.gov (United States)

    Jiang, Weidong; Mao, Ying Qing; Huang, Ruochun; Duan, Chaohui; Xi, Yun; Yang, Kai; Huang, Ruo-Pan

    2014-01-31

    Dried blood spot samples (DBSS) on filter paper offer several advantages compared to conventional serum/plasma samples: they do not require any phlebotomy or separation of blood by centrifugation; they are less invasive; they allow sample stability and shipment at room temperature; and they pose a negligible risk of infection with blood-borne viruses, such as HIV, HBV and HCV, to those who handle them. Therefore dried blood spot samples (DBSS) on filter paper can be a quick, convenient and inexpensive means of obtaining blood samples for biomarker discovery, disease screening, diagnosis and treatment monitoring in non-hospitalized, public health settings. In this study, we investigated for the first time the potential application of dried blood spot samples (DBSS) in protein expression profiling using antibody array technology. First, optimal conditions for array assay performance using dried blood spot samples (DBSS) was established, including sample elution buffer, elution time, elution temperature and assay blocking buffer. Second, we analyzed dried blood spot samples (DBSS) using three distinct antibody array platforms, including sandwich-based antibody arrays, quantitative antibody arrays and biotin-label-based antibody arrays. In comparison with paired serum samples, detection of circulating proteins in dried blood spot samples (DBSS) correlated well for both low- and high-abundance proteins on all three antibody array platforms. In conclusion, our study strongly indicates the novel application of multiplex antibody array platforms to analyze dried blood spot samples (DBSS) on filter paper represents a viable, cost-effective method for protein profiling, biomarker discovery and disease screening in a large, population-based survey. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Aflatoxin B1-lysine adduct in dried blood spot samples of animals and humans.

    Science.gov (United States)

    Xue, Kathy S; Cai, Wenjie; Tang, Lili; Wang, Jia-Sheng

    2016-12-01

    Dried blood spots (DBS) were proposed as potentially viable method for exposure assessment of environmental toxicants in infant and young children. For this study, we validated an experimental protocol to quantify AFB 1 -lysine adduct in DBS samples of AFB 1 -treated F344 rats, as well as samples from human field study. Significant dose-response relationships in AFB 1 -lysine adduct formation were found in DBS samples of rats treated with single- and repeated-dose AFB 1 . AFB 1 -lysine levels in DBS samples were highly correlated with corresponding serum sample levels. The Person coefficients were 0.997 for the single-dose exposure, and 0.996 for the repeated-dose exposure. Levels of AFB 1 -lysine adduct had also good agreement between DBS and serum samples as shown by Bland-Altman plot analysis. For human field study samples (n = 36), a Pearson correlation coefficient of 0.784 was found between AFB 1 -lysine adduct levels of DBS and corresponding serum samples. Bland-Altman plots showed the distribution of the log differences between DBS and serum AFB 1 -lysine levels are within 95% confidence intervals. These results showed AFB 1 -lysine adduct levels in DBS cards and serum samples from animals and human samples are comparable, and the DBS technique and analytical protocol is a good means to assess AFB 1 exposure in infant and children populations. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Feasibility of self-sampled dried blood spot and saliva samples sent by mail in a population-based study.

    Science.gov (United States)

    Sakhi, Amrit Kaur; Bastani, Nasser Ezzatkhah; Ellingjord-Dale, Merete; Gundersen, Thomas Erik; Blomhoff, Rune; Ursin, Giske

    2015-04-11

    In large epidemiological studies it is often challenging to obtain biological samples. Self-sampling by study participants using dried blood spots (DBS) technique has been suggested to overcome this challenge. DBS is a type of biosampling where blood samples are obtained by a finger-prick lancet, blotted and dried on filter paper. However, the feasibility and efficacy of collecting DBS samples from study participants in large-scale epidemiological studies is not known. The aim of the present study was to test the feasibility and response rate of collecting self-sampled DBS and saliva samples in a population-based study of women above 50 years of age. We determined response proportions, number of phone calls to the study center with questions about sampling, and quality of the DBS. We recruited women through a study conducted within the Norwegian Breast Cancer Screening Program. Invitations, instructions and materials were sent to 4,597 women. The data collection took place over a 3 month period in the spring of 2009. Response proportions for the collection of DBS and saliva samples were 71.0% (3,263) and 70.9% (3,258), respectively. We received 312 phone calls (7% of the 4,597 women) with questions regarding sampling. Of the 3,263 individuals that returned DBS cards, 3,038 (93.1%) had been packaged and shipped according to instructions. A total of 3,032 DBS samples were sufficient for at least one biomarker analysis (i.e. 92.9% of DBS samples received by the laboratory). 2,418 (74.1%) of the DBS cards received by the laboratory were filled with blood according to the instructions (i.e. 10 completely filled spots with up to 7 punches per spot for up to 70 separate analyses). To assess the quality of the samples, we selected and measured two biomarkers (carotenoids and vitamin D). The biomarker levels were consistent with previous reports. Collecting self-sampled DBS and saliva samples through the postal services provides a low cost, effective and feasible

  15. Feasibility of self-sampled dried blood spot and saliva samples sent by mail in a population-based study

    International Nuclear Information System (INIS)

    Sakhi, Amrit Kaur; Bastani, Nasser Ezzatkhah; Ellingjord-Dale, Merete; Gundersen, Thomas Erik; Blomhoff, Rune; Ursin, Giske

    2015-01-01

    In large epidemiological studies it is often challenging to obtain biological samples. Self-sampling by study participants using dried blood spots (DBS) technique has been suggested to overcome this challenge. DBS is a type of biosampling where blood samples are obtained by a finger-prick lancet, blotted and dried on filter paper. However, the feasibility and efficacy of collecting DBS samples from study participants in large-scale epidemiological studies is not known. The aim of the present study was to test the feasibility and response rate of collecting self-sampled DBS and saliva samples in a population–based study of women above 50 years of age. We determined response proportions, number of phone calls to the study center with questions about sampling, and quality of the DBS. We recruited women through a study conducted within the Norwegian Breast Cancer Screening Program. Invitations, instructions and materials were sent to 4,597 women. The data collection took place over a 3 month period in the spring of 2009. Response proportions for the collection of DBS and saliva samples were 71.0% (3,263) and 70.9% (3,258), respectively. We received 312 phone calls (7% of the 4,597 women) with questions regarding sampling. Of the 3,263 individuals that returned DBS cards, 3,038 (93.1%) had been packaged and shipped according to instructions. A total of 3,032 DBS samples were sufficient for at least one biomarker analysis (i.e. 92.9% of DBS samples received by the laboratory). 2,418 (74.1%) of the DBS cards received by the laboratory were filled with blood according to the instructions (i.e. 10 completely filled spots with up to 7 punches per spot for up to 70 separate analyses). To assess the quality of the samples, we selected and measured two biomarkers (carotenoids and vitamin D). The biomarker levels were consistent with previous reports. Collecting self-sampled DBS and saliva samples through the postal services provides a low cost, effective and feasible

  16. Robustness of genome-wide scanning using archived dried blood spot samples as a DNA source

    Directory of Open Access Journals (Sweden)

    Børglum Anders D

    2011-07-01

    Full Text Available Abstract Background The search to identify disease-susceptible genes requires access to biological material from numerous well-characterized subjects. Archived residual dried blood spot (DBS samples, also known as Guthrie cards, from national newborn screening programs may provide a DNA source for entire populations. Combined with clinical information from medical registries, DBS samples could provide a rich source for productive research. However, the amounts of DNA which can be extracted from these precious samples are minute and may be prohibitive for numerous genotypings. Previously, we demonstrated that DBS DNA can be whole-genome amplified and used for reliable genetic analysis on different platforms, including genome-wide scanning arrays. However, it remains unclear whether this approach is workable on a large sample scale. We examined the robustness of using DBS samples for whole-genome amplification following genome-wide scanning, using arrays from Illumina and Affymetrix. Results This study is based on 4,641 DBS samples from the Danish Newborn Screening Biobank, extracted for three separate genome-wide association studies. The amount of amplified DNA was significantly (P Conclusion Our study indicates that archived DBS samples from the Danish Newborn Screening Biobank represent a reliable resource of DNA for whole-genome amplification and subsequent genome-wide association studies. With call-rates equivalent to high quality DNA samples, our results point to new opportunities for using the neonatal biobanks available worldwide in the hunt for genetic components of disease.

  17. Creatinine measurement on dry blood spot sample for chronic kidney disease screening.

    Science.gov (United States)

    Silva, Alan Castro Azevedo E; Gómez, Juan Fidel Bencomo; Lugon, Jocemir Ronaldo; Graciano, Miguel Luis

    2016-03-01

    Chronic kidney disease (CKD) screening is advisable due to its high morbidity and mortality and is usually performed by sampling blood and urine. Here we present an innovative and simpler method, by measuring creatinine on a dry blood spot on filter paper. One-hundred and six individuals at high risk for CKD were enrolled. The creatinine values obtained using both tests and the demographic data of each participant allowed us to determinate the eGFR. The adopted cutoff for CKD was an eGFR value 96%, predictive negative value 55% and accuracy 92%. By the CKD-EPI equation the sensitivity was 94%, specificity 55%, predictive positive value 94%, predictive negative value 55% and accuracy 90%. A Bland and Altman analysis showed a relatively narrow range of creatinine values differences (+ 0.68mg/dl to -0.55mg/dl) inside the ± 1.96 SD, without systematic differences. Measurement of creatinine on dry blood sample is an easily feasible non-invasive diagnostic test with good accuracy that may be useful to screen chronic kidney disease.

  18. Robustness of genome-wide scanning using archived dried blood spot samples as a DNA source.

    Science.gov (United States)

    Hollegaard, Mads V; Grove, Jakob; Grauholm, Jonas; Kreiner-Møller, Eskil; Bønnelykke, Klaus; Nørgaard, Mette; Benfield, Thomas L; Nørgaard-Pedersen, Bent; Mortensen, Preben B; Mors, Ole; Sørensen, Henrik T; Harboe, Zitta B; Børglum, Anders D; Demontis, Ditte; Ørntoft, Torben F; Bisgaard, Hans; Hougaard, David M

    2011-07-04

    The search to identify disease-susceptible genes requires access to biological material from numerous well-characterized subjects. Archived residual dried blood spot (DBS) samples, also known as Guthrie cards, from national newborn screening programs may provide a DNA source for entire populations. Combined with clinical information from medical registries, DBS samples could provide a rich source for productive research. However, the amounts of DNA which can be extracted from these precious samples are minute and may be prohibitive for numerous genotypings. Previously, we demonstrated that DBS DNA can be whole-genome amplified and used for reliable genetic analysis on different platforms, including genome-wide scanning arrays. However, it remains unclear whether this approach is workable on a large sample scale. We examined the robustness of using DBS samples for whole-genome amplification following genome-wide scanning, using arrays from Illumina and Affymetrix. This study is based on 4,641 DBS samples from the Danish Newborn Screening Biobank, extracted for three separate genome-wide association studies. The amount of amplified DNA was significantly (P Biobank represent a reliable resource of DNA for whole-genome amplification and subsequent genome-wide association studies. With call-rates equivalent to high quality DNA samples, our results point to new opportunities for using the neonatal biobanks available worldwide in the hunt for genetic components of disease.

  19. Bridging the Gap between Sample Collection and Laboratory Analysis: Using Dried Blood Spots to Identify Human Exposure to Chemical Agents.

    Science.gov (United States)

    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.

  20. Bridging the gap between sample collection and laboratory analysis: using dried blood spots to identify human exposure to chemical agents

    Science.gov (United States)

    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.

  1. Effects of blood sample handling procedures on measurable inflammatory markers in plasma, serum and dried blood spot samples

    DEFF Research Database (Denmark)

    Skogstrand, K.; Thorsen, P.; Vogel, I.

    2008-01-01

    , and 30 days at the same temperatures. 27 inflammatory markers in serum and plasma and 25 markers in DBSS were measured by a previously validated multiplex sandwich immunoassay using Luminex xMAP technology. The measurable concentrations of several cytokines in serum and plasma were significantly......The interests in monitoring inflammation by immunoassay determination of blood inflammatory markers call for information on the stability of these markers in relation to the handling of blood samples. The increasing use of stored biobank samples for such ventures that may have been collected...... increased when blood samples were stored for a period of time before the centrifugation, for certain cytokines more than 1000 fold compared to serum and plasma isolated and frozen immediately after venepuncture. The concentrations in serum generally increased more than in plasma. The measurable...

  2. Evaluation of dried blood spot samples for hepatitis C virus detection and quantification.

    Science.gov (United States)

    Marques, Brunna Lemos Crespo; do Espírito-Santo, Márcia Paschoal; Marques, Vanessa Alves; Miguel, Juliana Custódio; da Silva, Elisangela Ferreira; Villela-Nogueira, Cristiane Alves; Lewis-Ximenez, Lia Laura; Lampe, Elisabeth; Villar, Livia Melo

    2016-09-01

    Dried blood spots (DBS) could be an excellent alternative for HCV diagnosis, since it is less invasive and can be stored and transported without refrigeration. The aim of this study was to optimize quantitative and qualitative methods for HCV detection in DBS. DBS and serum samples were collected from 99 subjects (59 anti-HCV/HCV RNA positive and 40 seronegative samples). Seven extraction methods and different PCR parameters were evaluated in DBS samples in the quantitative RT-PCR (qRT-PCR) developed to amplify the 5' noncoding region of HCV. A qualitative PCR for amplification of NS5B region of HCV was also valued and the nested-PCR sequenced. The qRT-PCR showed good correlation to commercial assay for HCV viral measurement in serum. To quantify HCV RNA in DBS, it was necessary to increase reverse transcriptase and cDNA concentration. HCV RNA quantification in DBS demonstrated sensitivity of 65.9%, 100% of specificity and kappa statistic of 0.65. The median viral load of DBS samples was 5.38 log10 copies/ml (minimum value=1.76 and maximum value=10.48 log10 copies/ml). HCV RNA was detected in NS5B regions and nucleotide sequences obtained in 43 serum and 11 DBS samples. The presence of the same subtype was observed in paired serum and DBS samples. In this study, it was possible to demonstrate that, despite the low sensitivity, the optimized protocol was able to determine the viral load, as well as, the infecting HCV genotype, validating the usefulness of DBS for viral load determination and molecular epidemiology studies of HCV. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Adiponectin levels measured in dried blood spot samples from neonates born small and appropriate for gestational age

    DEFF Research Database (Denmark)

    Klamer, A; Skogstrand, Kristin; Hougaard, D M

    2007-01-01

    Adiponectin levels measured in neonatal dried blood spot samples (DBSS) might be affected by both prematurity and being born small for gestational age (SGA). The aim of the study was to measure adiponectin levels in routinely collected neonatal DBSS taken on day 5 (range 3-12) postnatal from...

  4. Quantitative determination of atenolol in dried blood spot samples by LC-HRMS: a potential method for assessing medication adherence.

    Science.gov (United States)

    Lawson, Graham; Cocks, Elizabeth; Tanna, Sangeeta

    2012-05-15

    The use of blood spot collection cards was investigated as a means of obtaining small volume samples for the quantification of therapeutic drugs for assessing medication adherence. A liquid chromatography-high resolution TOF mass spectrometry (LC-HRMS) method, based on the measurement at the accurate mass to charge ratio of the target analyte, was used to ensure specificity for atenolol in the dried blood spot (DBS) samples. A working method was developed and validated. For the preparation of DBS samples whole blood spiked with analyte was used to produce 30 μl blood spots on specimen collection cards. A 5mm disc was cut from the dried blood spot and extracted using methanol:water (60:40, v/v) containing the internal standard, atenolol-d(7). Extracts were vortexed, sonicated and then centrifuged. Gradient chromatographic elution was achieved using an Ascentis Express C18 100mm×2.1mm column and a mobile phase flow rate of 0.2 ml/min and the column oven temperature at 30 °C. MS detection was carried out in electrospray positive ion mode for target ions at accurate mass m/z 267.1703 for atenolol and 274.2143 for the IS. Drug extraction efficiency from spiked blood spots was demonstrated to be 96±5% and the drug was stable in DBS for at least 10 weeks. The developed LC-HRMS method was linear within the tested calibration range of 25-1500 ng/ml and validation showed the accuracy (relative error) and precision (coefficient of variation) values were within the pre-defined limits of ≤ 5% at all concentrations with a limit of quantification of 25 ng/ml. Factors with potential to affect drug quantification measurements such as the matrix effects, volume of blood applied onto the collection card and effect of different sampling cards were investigated. The developed LC-HRMS method was applied to blood spots on sampling card taken from adult healthy volunteers previously administered a 50mg atenolol tablet and a DBS concentration-time profile was obtained for atenolol

  5. Quantification of rifapentine, a potent antituberculosis drug, from dried blood spot samples using liquid chromatographic-tandem mass spectrometric analysis.

    Science.gov (United States)

    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. Copyright © 2014

  6. Simplifying sample pretreatment: application of dried blood spot (DBS) method to blood samples, including postmortem, for UHPLC-MS/MS analysis of drugs of abuse.

    Science.gov (United States)

    Odoardi, Sara; Anzillotti, Luca; Strano-Rossi, Sabina

    2014-10-01

    The complexity of biological matrices, such as blood, requires the development of suitably selective and reliable sample pretreatment procedures prior to their instrumental analysis. A method has been developed for the analysis of drugs of abuse and their metabolites from different chemical classes (opiates, methadone, fentanyl and analogues, cocaine, amphetamines and amphetamine-like substances, ketamine, LSD) in human blood using dried blood spot (DBS) and subsequent UHPLC-MS/MS analysis. DBS extraction required only 100μL of sample, added with the internal standards and then three droplets (30μL each) of this solution were spotted on the card, let dry for 1h, punched and extracted with methanol with 0.1% of formic acid. The supernatant was evaporated and the residue was then reconstituted in 100μL of water with 0.1% of formic acid and injected in the UHPLC-MS/MS system. The method was validated considering the following parameters: LOD and LOQ, linearity, precision, accuracy, matrix effect and dilution integrity. LODs were 0.05-1ng/mL and LOQs were 0.2-2ng/mL. The method showed satisfactory linearity for all substances, with determination coefficients always higher than 0.99. Intra and inter day precision, accuracy, matrix effect and dilution integrity were acceptable for all the studied substances. The addition of internal standards before DBS extraction and the deposition of a fixed volume of blood on the filter cards ensured the accurate quantification of the analytes. The validated method was then applied to authentic postmortem blood samples. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  7. Comparative value of blood and skin samples for diagnosis of spotted fever group rickettsial infection in model animals.

    Science.gov (United States)

    Levin, Michael L; Snellgrove, Alyssa N; Zemtsova, Galina E

    2016-07-01

    The definitive diagnosis of spotted fever group (SFG) rickettsioses in humans is challenging due to the retrospective nature and cross reactivity of the serological methods and the absence of reliable and consistent samples for molecular diagnostics. Existing data indicate the transient character of bacteremia in experimentally infected animals. The ability of arthropod vectors to acquire rickettsial infection from the laboratory animals in the absence of systemic infection and known tropism of rickettsial agents to endothelial cells of peripheral blood vessels underline the importance of local infection and consequently the diagnostic potential of skin samples. In order to evaluate the diagnostic sensitivity of rickettsial DNA detection in blood and skin samples, we compared results of PCR testing in parallel samples collected from model laboratory animals infected with Rickettsia rickettsii, Rickettsia parkeri and Rickettsia slovaca-like agent at different time points after infection. Skin samples were collected from ears - away from the site of tick placement and without eschars. Overall, testing of skin samples resulted in a higher proportion of positive results than testing of blood samples. Presented data from model animals demonstrates that testing of skin samples from sites of rickettsial proliferation can provide definitive molecular diagnosis of up to 60-70% of tick-borne SFG rickettsial infections during the acute stage of illness. Detection of pathogen DNA in cutaneous samples is a valuable alternative to blood-PCR at least in model animals. Published by Elsevier GmbH.

  8. Evaluation of a novel dried blood spot collection device (HemaSpot™) to test blood samples collected from dogs for antibodies to Leishmania infantum.

    Science.gov (United States)

    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

  9. Direct analysis of site-specific N-glycopeptides of serological proteins in dried blood spot samples.

    Science.gov (United States)

    Choi, Na Young; Hwang, Heeyoun; Ji, Eun Sun; Park, Gun Wook; Lee, Ju Yeon; Lee, Hyun Kyoung; Kim, Jin Young; Yoo, Jong Shin

    2017-08-01

    Dried blood spot (DBS) samples have a number of advantages, especially with respect to ease of collection, transportation, and storage and to reduce biohazard risk. N-glycosylation is a major post-translational modification of proteins in human blood that is related to a variety of biological functions, including metastasis, cell-cell interactions, inflammation, and immunization. Here, we directly analyzed tryptic N-glycopeptides from glycoproteins in DBS samples using liquid chromatography-tandem mass spectrometry (LC-MS/MS) without centrifugation of blood samples, depletion of major proteins, desalting of tryptic peptides, and enrichment of N-glycopeptides. Using this simple method, we identified a total of 41 site-specific N-glycopeptides from 16 glycoproteins in the DBS samples, from immunoglobulin gamma 1 (IgG-1, 10 mg/mL) down to complement component C7 (50 μg/mL). Of these, 32 N-glycopeptides from 14 glycoproteins were consistently quantified over 180 days stored at room temperature. The major abundant glycoproteins in the DBS samples were IgG-1 and IgG-2, which contain nine asialo-fucosylated complex types of 16 different N-glycopeptide isoforms. Sialo-non-fucosylated complex types were primarily detected in the other glycoproteins such as alpha-1-acid glycoprotein 1, 2, alpha-1-antitypsin, alpha-2-macroglobulin, haptoglobin, hemopexin, Ig alpha 1, 2 chain C region, kininogen-1, prothrombin, and serotransferrin. We first report the characterization of site-specific N-glycoproteins in DBS samples by LC-MS/MS with minimal sample preparation.

  10. Evaluation of HBsAg and anti-HBc assays in saliva and dried blood spot samples according HIV status.

    Science.gov (United States)

    Flores, Geane Lopes; Cruz, Helena Medina; Potsch, Denise Vigo; May, Silvia Beatriz; Brandão-Mello, Carlos Eduardo; Pires, Marcia Maria Amendola; Pilotto, Jose Henrique; Lewis-Ximenez, Lia Laura; Lampe, Elisabeth; Villar, Livia Melo

    2017-09-01

    Influence of HIV status in HBV markers detection in saliva and dried blood spots (DBS) was not well established. This study aims to evaluate the performance of optimized commercial immunoassay for identifying HBsAg and anti-HBc in saliva and DBS according HIV status. A sum of 535 individuals grouped as HIV + , HBV + , HIV/HBV + and HIV/HBV- were recruited where 347 and 188 were included for HBsAg and anti-HBc evaluation, respectively. Serum, DBS collected in Whatman 903 paper and saliva obtained using salivette device were analyzed using EIA. Increased sample volume and ROC curve analysis for cut off determination were used for DBS and saliva testing. HBsAg detection in saliva and DBS exhibited sensitivities of 80.9% and 85.6% and specificities of 86.8% and 96.3%. Sensitivity of anti-HBc in saliva and DBS were 82.4% and 76.9% and specificities in saliva and DBS were 96.9% and 91.7%. Low sensitivities were observed for HBsAg (62%) and anti-HBc (47%) detection in saliva of HIV/HBV+ individuals. OD values were also lower for HBsAg detection in DBS and saliva of HIV/HBV+ individuals compared to their serum samples. Statistical significance was found for sensitivities in HBsAg detection between saliva and DBS demonstrating high sensitivity for DBS specimens. In conclusion, HIV status or antiretroviral treatment appears to interfere in the performance of HBsAg and anti-HBc detection in DBS and saliva samples using the adapted commercial EIA. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Archived neonatal dried blood spot samples can be used for accurate whole genome and exome-targeted next-generation sequencing

    DEFF Research Database (Denmark)

    Hollegaard, Mads Vilhelm; Grauholm, Jonas; Nielsen, Ronni

    2013-01-01

    Dried blood spot samples (DBSS) have been collected and stored for decades as part of newborn screening programmes worldwide. Representing almost an entire population under a certain age and collected with virtually no bias, the Newborn Screening Biobanks are of immense value in medical studies......, for example, to examine the genetics of various disorders. We have previously demonstrated that DNA extracted from a fraction (2×3.2mm discs) of an archived DBSS can be whole genome amplified (wgaDNA) and used for accurate array genotyping. However, until now, it has been uncertain whether wgaDNA from DBSS...... can be used for accurate whole genome sequencing (WGS) and exome sequencing (WES). This study examined two individuals represented by three different types of samples each: whole-blood (reference samples), 3-year-old DBSS spotted with reference material (refDBSS), and 27- to 29-year-old archived...

  12. Study of measurement of the alcohol biomarker phosphatidylethanol (PEth) in dried blood spot (DBS) samples and application of a volumetric DBS device.

    Science.gov (United States)

    Beck, Olof; Kenan Modén, Naama; Seferaj, Sabina; Lenk, Gabriel; Helander, Anders

    2018-04-01

    Phosphatidylethanol (PEth) is a group of phospholipids formed in cell membranes following alcohol consumption. PEth measurement in whole blood samples is established as a specific alcohol biomarker with clinical and medico-legal applications. This study further evaluated the usefulness of dried blood spot (DBS) samples collected on filter paper for PEth measurement. Specimens used were surplus volumes of venous whole blood sent for routine LC-MS/MS quantification of PEth 16:0/18:1, the major PEth homolog. DBS samples were prepared by pipetting blood on Whatman 903 Protein Saver Cards and onto a volumetric DBS device (Capitainer). The imprecision (CV) of the DBS sample amount based on area and weight measurements of spot punches were 23-28%. Investigation of the relationship between blood hematocrit and PEth concentration yielded a linear, positive correlation, and at around 1.0-1.5μmol/L PEth 16:0/18:1, the PEth concentration increased by ~0.1μmol/L for every 5% increase in hematocrit. There was a close agreement between the PEth concentrations obtained with whole blood samples and the corresponding results using Whatman 903 (PEth DBS =1.026 PEth WB +0.013) and volumetric device (PEth DBS =1.045 PEth WB +0.016) DBS samples. The CV of PEth quantification in DBS samples at concentrations≥0.05μmol/L were ≤15%. The present results further confirmed the usefulness of DBS samples for PEth measurement. Copyright © 2018 Elsevier B.V. All rights reserved.

  13. Detecting 22q11.2 deletions by use of multiplex ligation-dependent probe amplification on DNA from neonatal dried blood spot samples

    DEFF Research Database (Denmark)

    Sørensen, Karina; Agergaard, Peter Juul; Olesen, Charlotte

    2010-01-01

    of 22q11.2 deletions among certain manifestations, eg, congenital heart disease, on selected Danes, a multiplex ligation-dependant probe amplification (MLPA) analysis was designed. The analysis was planned to be performed on DNA extracted from dried blood spot samples (DBSS) obtained from Guthrie cards...... probe design is successful and reliable using minimal amounts of DNA. This allows for use of DBSS samples in a retrospective study of 22q11.2 deletion among certain manifestations associated with DiGeorge Syndrome....

  14. Archived neonatal dried blood spot samples can be used for accurate whole genome and exome-targeted next-generation sequencing.

    Science.gov (United States)

    Hollegaard, Mads Vilhelm; Grauholm, Jonas; Nielsen, Ronni; Grove, Jakob; Mandrup, Susanne; Hougaard, David Michael

    2013-01-01

    Dried blood spot samples (DBSS) have been collected and stored for decades as part of newborn screening programmes worldwide. Representing almost an entire population under a certain age and collected with virtually no bias, the Newborn Screening Biobanks are of immense value in medical studies, for example, to examine the genetics of various disorders. We have previously demonstrated that DNA extracted from a fraction (2×3.2mm discs) of an archived DBSS can be whole genome amplified (wgaDNA) and used for accurate array genotyping. However, until now, it has been uncertain whether wgaDNA from DBSS can be used for accurate whole genome sequencing (WGS) and exome sequencing (WES). This study examined two individuals represented by three different types of samples each: whole-blood (reference samples), 3-year-old DBSS spotted with reference material (refDBSS), and 27- to 29-year-old archived neonatal DBSS (neoDBSS) stored at -20°C in the Danish Newborn Screening Biobank. The reference samples were genotyped using an Illumina Omni2.5M array, and all samples were sequenced on a HighSeq2000 Paired-End flow cell. First, we compared the array single nucleotide polymorphism (SNP) genotype data to the single nucleotide variation (SNV) calls from the WGS and WES SNV calls. We also compared the WGS and WES reference sample SNV calls to the DBSS SNV calls. The overall performance of the archived DBSS was similar to the whole blood reference sample. Plotting the error rates relative to coverage revealed that the error rates of DBSS were similar to that of their reference samples. SNVs called with a coverage<×8 had error rates between 1.5 and 35%, whereas the error rates of SNVs called with a coverage≥8 were <1.5%. In conclusion, the wgaDNA amplified from both new and old neonatal DBSS perform as well as their whole-blood reference samples with regards to error rates, strongly indicating that neonatal DBSS collected shortly after birth and stored for decades comprise an

  15. Application of dried blood spots to determine vitamin D status in a large nutritional study with unsupervised sampling: the Food4Me project.

    Science.gov (United States)

    Hoeller, Ulrich; Baur, Manuela; Roos, Franz F; Brennan, Lorraine; Daniel, Hannelore; Fallaize, Rosalind; Forster, Hannah; Gibney, Eileen R; Gibney, Mike; Godlewska, Magdalena; Hartwig, Kai; Kolossa, Silvia; Lambrinou, Christina P; Livingstone, Katherine M; Lovegrove, Julie A; Macready, Anna L; Manios, Yannis; Marsaux, Cyril F M; Martinez, J Alfredo; Celis-Morales, Carlos; Moschonis, George; Navas-Carretero, Santiago; O'Donovan, Clare B; San-Cristobal, Rodrigo; Saris, Wim H M; Surwiłło, Agnieszka; Traczyk, Iwona; Tsirigoti, Lydia; Walsh, Marianne C; Woolhead, Clara; Mathers, John C; Weber, Peter

    2016-01-28

    An efficient and robust method to measure vitamin D (25-hydroxy vitamin D3 (25(OH)D3) and 25-hydroxy vitamin D2 in dried blood spots (DBS) has been developed and applied in the pan-European multi-centre, internet-based, personalised nutrition intervention study Food4Me. The method includes calibration with blood containing endogenous 25(OH)D3, spotted as DBS and corrected for haematocrit content. The methodology was validated following international standards. The performance characteristics did not reach those of the current gold standard liquid chromatography-MS/MS in plasma for all parameters, but were found to be very suitable for status-level determination under field conditions. DBS sample quality was very high, and 3778 measurements of 25(OH)D3 were obtained from 1465 participants. The study centre and the season within the study centre were very good predictors of 25(OH)D3 levels (Pvitamin D status information in a large nutritional study.

  16. A simple assay of paracetamol based on dried blood spot suitable ...

    African Journals Online (AJOL)

    Dried blood spots in Guthrie cards are a reliable means of blood sampling suitable for pharmacoki-netic analysis in children. The aim of this study was to develop a simple and reliable bioanalytical method to measure the concentration of paracetamol in dried blood spots. Paracetamol was ex-tracted from dry blood spots by ...

  17. Diagnostic accuracy of serological diagnosis of hepatitis C and B using dried blood spot samples (DBS: two systematic reviews and meta-analyses

    Directory of Open Access Journals (Sweden)

    Berit Lange

    2017-11-01

    Full Text Available Abstract Background Dried blood spots (DBS are a convenient tool to enable diagnostic testing for viral diseases due to transport, handling and logistical advantages over conventional venous blood sampling. A better understanding of the performance of serological testing for hepatitis C (HCV and hepatitis B virus (HBV from DBS is important to enable more widespread use of this sampling approach in resource limited settings, and to inform the 2017 World Health Organization (WHO guidance on testing for HBV/HCV. Methods We conducted two systematic reviews and meta-analyses on the diagnostic accuracy of HCV antibody (HCV-Ab and HBV surface antigen (HBsAg from DBS samples compared to venous blood samples. MEDLINE, EMBASE, Global Health and Cochrane library were searched for studies that assessed diagnostic accuracy with DBS and agreement between DBS and venous sampling. Heterogeneity of results was assessed and where possible a pooled analysis of sensitivity and specificity was performed using a bivariate analysis with maximum likelihood estimate and 95% confidence intervals (95%CI. We conducted a narrative review on the impact of varying storage conditions or limits of detection in subsets of samples. The QUADAS-2 tool was used to assess risk of bias. Results For the diagnostic accuracy of HBsAg from DBS compared to venous blood, 19 studies were included in a quantitative meta-analysis, and 23 in a narrative review. Pooled sensitivity and specificity were 98% (95%CI:95%–99% and 100% (95%CI:99–100%, respectively. For the diagnostic accuracy of HCV-Ab from DBS, 19 studies were included in a pooled quantitative meta-analysis, and 23 studies were included in a narrative review. Pooled estimates of sensitivity and specificity were 98% (CI95%:95–99 and 99% (CI95%:98–100, respectively. Overall quality of studies and heterogeneity were rated as moderate in both systematic reviews. Conclusion HCV-Ab and HBsAg testing using DBS compared to venous

  18. Diagnostic accuracy of serological diagnosis of hepatitis C and B using dried blood spot samples (DBS): two systematic reviews and meta-analyses.

    Science.gov (United States)

    Lange, Berit; Cohn, Jennifer; Roberts, Teri; Camp, Johannes; Chauffour, Jeanne; Gummadi, Nina; Ishizaki, Azumi; Nagarathnam, Anupriya; Tuaillon, Edouard; van de Perre, Philippe; Pichler, Christine; Easterbrook, Philippa; Denkinger, Claudia M

    2017-11-01

    Dried blood spots (DBS) are a convenient tool to enable diagnostic testing for viral diseases due to transport, handling and logistical advantages over conventional venous blood sampling. A better understanding of the performance of serological testing for hepatitis C (HCV) and hepatitis B virus (HBV) from DBS is important to enable more widespread use of this sampling approach in resource limited settings, and to inform the 2017 World Health Organization (WHO) guidance on testing for HBV/HCV. We conducted two systematic reviews and meta-analyses on the diagnostic accuracy of HCV antibody (HCV-Ab) and HBV surface antigen (HBsAg) from DBS samples compared to venous blood samples. MEDLINE, EMBASE, Global Health and Cochrane library were searched for studies that assessed diagnostic accuracy with DBS and agreement between DBS and venous sampling. Heterogeneity of results was assessed and where possible a pooled analysis of sensitivity and specificity was performed using a bivariate analysis with maximum likelihood estimate and 95% confidence intervals (95%CI). We conducted a narrative review on the impact of varying storage conditions or limits of detection in subsets of samples. The QUADAS-2 tool was used to assess risk of bias. For the diagnostic accuracy of HBsAg from DBS compared to venous blood, 19 studies were included in a quantitative meta-analysis, and 23 in a narrative review. Pooled sensitivity and specificity were 98% (95%CI:95%-99%) and 100% (95%CI:99-100%), respectively. For the diagnostic accuracy of HCV-Ab from DBS, 19 studies were included in a pooled quantitative meta-analysis, and 23 studies were included in a narrative review. Pooled estimates of sensitivity and specificity were 98% (CI95%:95-99) and 99% (CI95%:98-100), respectively. Overall quality of studies and heterogeneity were rated as moderate in both systematic reviews. HCV-Ab and HBsAg testing using DBS compared to venous blood sampling was associated with excellent diagnostic accuracy

  19. Analysis of blood spots for polyfluoroalkyl chemicals

    Energy Technology Data Exchange (ETDEWEB)

    Kato, Kayoko; Wanigatunga, Amal A.; Needham, Larry L. [Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA (United States); Calafat, Antonia M., E-mail: acalafat@cdc.gov [Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA (United States)

    2009-12-10

    Polyfluoroalkyl chemicals (PFCs) have been detected in humans, in the environment, and in ecosystems around the world. The potential for developmental and reproductive toxicities of some PFCs is of concern especially to children's health. In the United States, a sample of a baby's blood, called a 'dried blood spot' (DBS), is obtained from a heel stick within 48 h of a child's birth. DBS could be useful for assessing prenatal exposure to PFCs. We developed a method based on online solid phase extraction coupled with high performance liquid chromatography-isotope dilution tandem mass spectrometry for measuring four PFCs in DBS, perfluorooctane sulfonate (PFOS), perfluorohexane sulfonate, perfluorooctanoate (PFOA), and perfluorononanoate. The analytical limits of detection using one whole DBS ({approx}75 {mu}L of blood) were <0.5 ng mL{sup -1}. To validate the method, we analyzed 98 DBS collected in May 2007 in the United States. PFOS and PFOA were detected in all DBS at concentrations in the low ng mL{sup -1} range. These data suggest that DBS may be a suitable matrix for assessing perinatal exposure to PFCs, but additional information related to sampling and specimen storage is needed to demonstrate the utility of these measures for assessing exposure.

  20. Automated high-capacity on-line extraction and bioanalysis of dried blood spot samples using liquid chromatography/high-resolution accurate mass spectrometry.

    Science.gov (United States)

    Oliveira, Regina V; Henion, Jack; Wickremsinhe, Enaksha R

    2014-11-30

    Pharmacokinetic data to support clinical development of pharmaceuticals are routinely obtained from liquid plasma samples. The plasma samples require frozen shipment and storage and are extracted off-line from the liquid chromatography/tandem mass spectrometry (LC/MS/MS) systems. In contrast, the use of dried blood spot (DBS) sampling is an attractive alternative in part due to its benefits in microsampling as well as simpler sample storage and transport. However, from a practical aspect, sample extraction from DBS cards can be challenging as currently performed. The goal of this report was to integrate automated serial extraction of large numbers of DBS cards with on-line liquid chromatography/high-resolution accurate mass spectrometry (LC/HRAMS) bioanalysis. An automated system for direct DBS extraction coupled to a LC/HRAMS was employed for the quantification of midazolam (MDZ) and α-hydroxymidazolam (α-OHMDZ) in human blood. The target analytes were directly extracted from the DBS cards onto an on-line chromatographic guard column followed by HRAMS detection. No additional sample treatment was required. The automated DBS LC/HRAMS method was developed and validated, based on the measurement at the accurate mass-to-charge ratio of the target analytes to ensure specificity for the assay. The automated DBS LC/HRAMS method analyzed a DBS sample within 2 min without the need for punching or additional off-line sample treatment. The fully automated analytical method was shown to be sensitive and selective over the concentration range of 5 to 2000 ng/mL. Intra- and inter-day precision and accuracy was less than 15% (less than 20% at the LLOQ). The validated method was successfully applied to measure MDZ and α-OHMDZ in an incurred human sample after a single 7.5 mg dose of MDZ. The direct DBS LC/HRAMS method demonstrated successful implementation of automated DBS extraction and bioanalysis for MDZ and α-OHMDZ. This approach has the potential to promote workload

  1. Diagnostic accuracy of detection and quantification of HBV-DNA and HCV-RNA using dried blood spot (DBS) samples - a systematic review and meta-analysis.

    Science.gov (United States)

    Lange, Berit; Roberts, Teri; Cohn, Jennifer; Greenman, Jamie; Camp, Johannes; Ishizaki, Azumi; Messac, Luke; Tuaillon, Edouard; van de Perre, Philippe; Pichler, Christine; Denkinger, Claudia M; Easterbrook, Philippa

    2017-11-01

    The detection and quantification of hepatitis B (HBV) DNA and hepatitis C (HCV) RNA in whole blood collected on dried blood spots (DBS) may facilitate access to diagnosis and treatment of HBV and HCV infection in resource-poor settings. We evaluated the diagnostic performance of DBS compared to venous blood samples for detection and quantification of HBV-DNA and HCV-RNA in two systematic reviews and meta-analyses on the diagnostic accuracy of HBV DNA and HCV RNA from DBS compared to venous blood samples. We searched MEDLINE, Embase, Global Health, Web of Science, LILAC and Cochrane library for studies that assessed diagnostic accuracy with DBS. Heterogeneity was assessed and where appropriate pooled estimates of sensitivity and specificity were generated using bivariate analyses with maximum likelihood estimates and 95% confidence intervals. We also conducted a narrative review on the impact of varying storage conditions or different cut-offs for detection from studies that undertook this in a subset of samples. The QUADAS-2 tool was used to assess risk of bias. In the quantitative synthesis for diagnostic accuracy of HBV-DNA using DBS, 521 citations were identified, and 12 studies met the inclusion criteria. Overall quality of studies was rated as low. The pooled estimate of sensitivity and specificity for HBV-DNA was 95% (95% CI: 83-99) and 99% (95% CI: 53-100), respectively. In the two studies that reported on cut-offs and limit of detection (LoD) - one reported a sensitivity of 98% for a cut-off of ≥2000 IU/ml and another reported a LoD of 914 IU/ml using a commercial assay. Varying storage conditions for individual samples did not result in a significant variation of results. In the synthesis for diagnostic accuracy of HCV-RNA using DBS, 15 studies met the inclusion criteria, and this included six additional studies to a previously published review. The pooled sensitivity and specificity was 98% (95% CI:95-99) and 98% (95% CI:95-99.0), respectively

  2. Comparing identified and statistically significant lipids and polar metabolites in 15-year old serum and dried blood spot samples for longitudinal studies: Comparing lipids and metabolites in serum and DBS samples

    Energy Technology Data Exchange (ETDEWEB)

    Kyle, Jennifer E. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Casey, Cameron P. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Stratton, Kelly G. [National Security Directorate, Pacific Northwest National Laboratory, Richland WA USA; Zink, Erika M. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Kim, Young-Mo [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Zheng, Xueyun [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Monroe, Matthew E. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Weitz, Karl K. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Bloodsworth, Kent J. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Orton, Daniel J. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Ibrahim, Yehia M. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Moore, Ronald J. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Lee, Christine G. [Department of Medicine, Bone and Mineral Unit, Oregon Health and Science University, Portland OR USA; Research Service, Portland Veterans Affairs Medical Center, Portland OR USA; Pedersen, Catherine [Department of Medicine, Bone and Mineral Unit, Oregon Health and Science University, Portland OR USA; Orwoll, Eric [Department of Medicine, Bone and Mineral Unit, Oregon Health and Science University, Portland OR USA; Smith, Richard D. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Burnum-Johnson, Kristin E. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA; Baker, Erin S. [Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland WA USA

    2017-02-05

    The use of dried blood spots (DBS) has many advantages over traditional plasma and serum samples such as smaller blood volume required, storage at room temperature, and ability for sampling in remote locations. However, understanding the robustness of different analytes in DBS samples is essential, especially in older samples collected for longitudinal studies. Here we analyzed DBS samples collected in 2000-2001 and stored at room temperature and compared them to matched serum samples stored at -80°C to determine if they could be effectively used as specific time points in a longitudinal study following metabolic disease. Four hundred small molecules were identified in both the serum and DBS samples using gas chromatograph-mass spectrometry (GC-MS), liquid chromatography-MS (LC-MS) and LC-ion mobility spectrometry-MS (LC-IMS-MS). The identified polar metabolites overlapped well between the sample types, though only one statistically significant polar metabolite in a case-control study was conserved, indicating degradation occurs in the DBS samples affecting quantitation. Differences in the lipid identifications indicated that some oxidation occurs in the DBS samples. However, thirty-six statistically significant lipids correlated in both sample types indicating that lipid quantitation was more stable across the sample types.

  3. Screening for HIV, hepatitis B and syphilis on dried blood spots: A promising method to better reach hidden high-risk populations with self-collected sampling.

    Directory of Open Access Journals (Sweden)

    Inge H M van Loo

    Full Text Available Many people at high risk for sexually transmitted infections (STIs, e.g., men who have sex with men (MSM, are not optimally reached by current sexual health care systems with testing. To facilitate testing by home-based sampling or sampling in outreach setting we evaluated dried blood spots (DBS, a method for self-collected blood sampling for serological screening of HIV, hepatitis B (HBV and syphilis. The aims of this study were to assess the acceptability and feasibility of self-collected DBS and to compare the test results for screening of HIV, HBV and syphilis from DBS with blood drawn by venous puncture.DBS were collected from men who have sex with men (MSM, visiting the STI clinic of the public health service South Limburg (n = 183 and HIV positive and HBV positive patients (n = 34, visiting the outpatient clinics of the Maastricht University Medical Centre in the period January 2012-April 2015. The 93 first participating MSM visiting the STI clinic were asked to fill in a questionnaire about the feasibility and acceptability about self-collection of DBS in a setting without going to a health care facility and were asked to collect the DBS themselves. Serological screening tests for HIV (HIV combi PT, Roche, HBV (HBsAg, Roche and syphilis (Treponema pallidum Ig, Biokit 3.0 were performed on DBS and on blood drawn by venous puncture, which was routinely taken for screening.In total 217 participants were included in the study with a median age of 40 years (range between 17-80. Of MSM 84% agreed that it was clear and easy to do the finger-prick, while 53% agreed that it was clear and easy to apply the blood onto the DBS card. Also, 80% of MSM would use the bloodspot test again. In 91% (198 of DBS, sufficient material was collected to perform the three tests. No difference was observed in DBS quality between self-collected DBS and health care worker collected DBS. For HIV (n = 195 DBS-serum pairs sensitivity and specificity were 100%. For HBV

  4. Use of dried-blood-spot samples and in-house assays to identify antiretroviral drug resistance in HIV-infected children in resource-constrained settings.

    Science.gov (United States)

    Ziemniak, Carrie; Mengistu, Yohannes; Ruff, Andrea; Chen, Ya-Hui; Khaki, Leila; Bedri, Abubaker; Simen, Birgitte B; Palumbo, Paul; Eshleman, Susan H; Persaud, Deborah

    2011-12-01

    Monitoring HIV drug resistance is an important component of the World Health Organization's global HIV program. HIV drug resistance testing is optimal with commercially available clinically validated test kits using plasma; however, that type of testing may not be feasible or affordable in resource-constrained settings. HIV genotyping from dried blood spots (DBS) with noncommercial (in-house) assays may facilitate the capture of HIV drug resistance outcomes in resource-constrained settings but has had varying rates of success. With in-house assays for HIV reverse transcriptase, we evaluated the yield of genotyping DBS samples collected from HIV-infected children who were enrolled in two clinical trials conducted in sub-Saharan Africa (median HIV viral load, 5.88 log(10) HIV RNA copies/ml; range, 4.04 to 6.99). Overall, HIV genotypes were obtained for 94 (89.5%) of 105 samples tested (95% and 84% from clinical trials #1 and #2, respectively); however, successful analysis of 15 (16.1%) of the 94 samples required repeat testing using a different set of primers on previously synthesized cDNA. The yield of genotyping was lower on the DBS that were stored suboptimally from clinical trial #2 (56% versus 88% for optimally stored). Concordance with plasma genotypes derived using a clinically validated, commercial kit-based assay (ViroSeq HIV-1 genotyping system) was also assessed in a subset of children with paired testing. For 34 samples with paired DBS and plasma genotypes, there was 100% concordance for major drug resistance mutations. DBS genotyping using in-house assays provides an alternative for antiretroviral drug resistance testing in children in resource-constrained regions but may require region-specific optimization before widespread use.

  5. Use of Dried-Blood-Spot Samples and In-House Assays To Identify Antiretroviral Drug Resistance in HIV-Infected Children in Resource-Constrained Settings ▿

    Science.gov (United States)

    Ziemniak, Carrie; Mengistu, Yohannes; Ruff, Andrea; Chen, Ya-Hui; Khaki, Leila; Bedri, Abubaker; Simen, Birgitte B.; Palumbo, Paul; Eshleman, Susan H.; Persaud, Deborah

    2011-01-01

    Monitoring HIV drug resistance is an important component of the World Health Organization's global HIV program. HIV drug resistance testing is optimal with commercially available clinically validated test kits using plasma; however, that type of testing may not be feasible or affordable in resource-constrained settings. HIV genotyping from dried blood spots (DBS) with noncommercial (in-house) assays may facilitate the capture of HIV drug resistance outcomes in resource-constrained settings but has had varying rates of success. With in-house assays for HIV reverse transcriptase, we evaluated the yield of genotyping DBS samples collected from HIV-infected children who were enrolled in two clinical trials conducted in sub-Saharan Africa (median HIV viral load, 5.88 log10 HIV RNA copies/ml; range, 4.04 to 6.99). Overall, HIV genotypes were obtained for 94 (89.5%) of 105 samples tested (95% and 84% from clinical trials #1 and #2, respectively); however, successful analysis of 15 (16.1%) of the 94 samples required repeat testing using a different set of primers on previously synthesized cDNA. The yield of genotyping was lower on the DBS that were stored suboptimally from clinical trial #2 (56% versus 88% for optimally stored). Concordance with plasma genotypes derived using a clinically validated, commercial kit-based assay (ViroSeq HIV-1 genotyping system) was also assessed in a subset of children with paired testing. For 34 samples with paired DBS and plasma genotypes, there was 100% concordance for major drug resistance mutations. DBS genotyping using in-house assays provides an alternative for antiretroviral drug resistance testing in children in resource-constrained regions but may require region-specific optimization before widespread use. PMID:21956987

  6. Performance of the biomerieux DBS puncher and dried blood spots ...

    African Journals Online (AJOL)

    Introduction: The latest World Health Organization recommendations request viral load (VL) testing, if possible, for monitoring HIV-1 infections. However, the use of plasma is an obstacle to realize this test in sub-Saharan Africa. In this context, the dried blood spot (DBS) is an interesting tool for sample collections.

  7. Mass spectrometric studies on the in vivo metabolism and excretion of SIRT1 activating drugs in rat urine, dried blood spots, and plasma samples for doping control purposes.

    Science.gov (United States)

    Höppner, Sebastian; Delahaut, Philippe; Schänzer, Wilhelm; Thevis, Mario

    2014-01-01

    The NAD(+) depending enzyme SIRT1 regulates the mitochondrial biogenesis, fat and glucose metabolism through catalyzing the deacetylation of several metabolism-related protein-substrates. Recently, synthetic activators of SIRT1 referred to as STACs (Sirtuin activating compounds, e.g. SRT2104) were identified and tested in clinical studies for the treatment of aging-related diseases such as type 2 diabetes, Alzheimer's and obesity. Although the mechanism of SIRT1 activation by small molecules has caused considerable controversy, STACs demonstrated a significant performance enhancement in mice experiments including an improvement of endurance, muscle strength, and locomotor behavior. Due to their potential to increase exercise tolerance in healthy individuals, SIRT1 activators are currently being monitored by anti-doping authorities. In the present study, the in vivo metabolic clearance of three SIRT1 activators was investigated in rats by the collection of urine, DBS (dried blood spots) and plasma samples following a single oral administration. The resulting metabolic products were studied by positive electrospray ionization - (tandem) mass spectrometry and confirmed by the comparison with in vitro generated metabolites using human and rat liver microsomal preparations. Subsequently, a screening procedure for five SIRT1 activators and the metabolite M1-SRT1720 in DBS specimens was developed. Liquid-liquid-extraction and liquid chromatography/tandem mass spectrometry was employed based on diagnostic ion transitions recorded in multiple reaction monitoring mode and two deuterated internal standards namely d8-SRT1720 and d8-M1-SRT1720 were utilized. The doping control assay was characterized with regard to specificity, limit of detection (10-50ng/ml), recovery (65-83%) and imprecision (7-20%) and ion suppression/enhancement effects (drug testing applications. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. Blood and dried blood spot telomere length measurement by qPCR: assay considerations.

    Directory of Open Access Journals (Sweden)

    DeAnna L Zanet

    Full Text Available Measurement of telomere length is crucial for the study of telomere maintenance and its role in molecular pathophysiology of diseases and in aging. Several methods are used to measure telomere length, the choice of which usually depends on the type and size of sample to be assayed, as well as cost and throughput considerations. The goal of this study was to investigate the factors that may influence the reliability of qPCR-based relative telomere length measurements in whole blood. Day to day intra-individual variability, types of blood anticoagulant, sample storage conditions, processing and site of blood draw were investigated. Two qPCR-based methods to measure telomere length (monoplex vs. multiplex were also investigated and showed a strong correlation between them. Freezing and thawing of the blood and storage of the blood at 4°C for up to 4 days did not affect telomere length values. Telomere lengths in dried blood spots were significantly higher than both whole blood and peripheral mononuclear blood cells, and were highly correlated with both. We found that telomere length measurements were significantly higher in dried blood spots collected directly from fingertip prick compared to dried blood spots prepared with anticoagulated whole blood collected from the finger, and non-blotted whole blood taken from both finger and arm venipuncture. This suggests that DNA from cells blotted on paper is not equivalent to that collected from venipuncture whole blood, and caution should be taken when comparing between blood sample types.

  9. A field trial of a PCR-based Mansonella ozzardi diagnosis assay detects high-levels of submicroscopic M. ozzardi infections in both venous blood samples and FTA card dried blood spots.

    Science.gov (United States)

    Medeiros, Jansen Fernandes; Almeida, Tatiana Amaral Pires; Silva, Lucyane Bastos Tavares; Rubio, Jose Miguel; Crainey, James Lee; Pessoa, Felipe Arley Costa; Luz, Sergio Luiz Bessa

    2015-05-20

    Mansonella ozzardi is a poorly understood human filarial parasite with a broad distribution throughout Latin America. Most of what is known about its parasitism has come from epidemiological studies that have estimated parasite incidence using light microscopy. Light microscopy can, however, miss lighter, submicroscopic, infections. In this study we have compared M. ozzardi incidence estimates made using light microscopy, with estimates made using PCR. 214 DNA extracts made from Large Volume Venous Blood Samples (LVVBS) were taken from volunteers from two study sites in the Rio Solimões region: Codajás [n = 109] and Tefé [n = 105] and were subsequently assayed for M. ozzardi parasitism using a diagnostic PCR (Mo-dPCR). Peripheral finger-prick blood samples were taken from the same individuals and used for microscopic examination. Finger-prick blood, taken from individuals from Tefé, was also used for the creation of FTAcard dried blood spots (DBS) that were subsequently subjected to Mo-dPCR. Overall M. ozzardi incidence estimates made with LVVBS PCRs were 1.8 times higher than those made using microscopy (44.9% [96/214] compared with 24.3% [52/214]) and 1.5 times higher than the PCR estimates made from FTAcard DBS (48/105 versus 31/105). PCR-based detection of FTAcard DBS proved 1.3 times more sensitive at diagnosing infections from peripheral blood samples than light microscopy did: detecting 24/105 compared with 31/105. PCR of LVVBS reported the fewest number of false negatives, detecting: 44 of 52 (84.6%) individuals diagnosed by microscopy; 27 of 31 (87.1%) of those diagnosed positive from DBSs and 17 out of 18 (94.4%) of those diagnosed as positive by both alternative methodologies. In this study, Mo-dPCR of LVVBS was by far the most sensitive method of detecting M. ozzardi infections and detected submicroscopic infections. Mo-dPCR FTAcard DBS also provided a more sensitive test for M. ozzardi diagnosis than light microscopy based diagnosis did and

  10. Analysis of ochratoxin A in dried blood spots - Correlation between venous and finger-prick blood, the influence of hematocrit and spotted volume.

    Science.gov (United States)

    Osteresch, Bernd; Cramer, Benedikt; Humpf, Hans-Ulrich

    2016-05-01

    We report the improvement of a method for the detection of ochratoxin A (OTA) and its thermal degradation product 2'R-ochratoxin A in dried blood spots (DBS) by high performance liquid chromatographic (HPLC) tandem mass spectrometry (MS/MS). The DBS technique was advanced for the analysis of these two compounds in DBS with unknown amounts of blood as well as varying hematocrit values. Furthermore the comparability of venous vs. capillary blood was investigated. Human whole blood samples were spotted, dried, and extracted with a solvent consisting of acetone, acetonitrile and water for analysis by HPLC-MS/MS. Quantification was carried out by stable isotope labelled internal standards. Blood samples of volunteers (n=50) were used to further optimize and simplify the procedure. Ochratoxin A and 2'R-ochratoxin A concentrations found in the entire spots (approx. 100 μL blood) were compared with punched DBS discs of 8.8mm size containing approximately 20 μL blood. As a result the amounts of both toxins in a punched 8.8mm disc correlate well with the entire DBS. Also the use of capillary blood from finger-pricks versus venous blood was evaluated. The analyte levels correlate as well indicating that the less invasive finger-prick sampling gives also reliable results. The influence of hematocrit was investigated in a range of 25-55% according to the hematocrit in the used real blood samples (34-46% hematocrit). However no significant hematocrit effect was observed for the utilized real blood samples. Moreover different blood volumes were spotted and punched as a minimal spot size is usually recommended for accurate analysis. In this experiment finger-prick samples typically consist of about 90 μL blood. Therefore spots of 75, 100 and 125 μL blood were prepared and analyzed. Similar to the hematocrit effect, no considerable influence was observed. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Quantitation of 5-Methyltetrahydrofolic Acid in Dried Blood Spots and Dried Plasma Spots by Stable Isotope Dilution Assays.

    Directory of Open Access Journals (Sweden)

    Markus Kopp

    Full Text Available Because of minimal data available on folate analysis in dried matrix spots (DMSs, we combined the advantages of stable isotope dilution assays followed by LC-MS/MS analysis with DMS sampling to develop a reliable method for the quantitation of plasma 5-methyltetrahydrofolic acid in dried blood spots (DBSs and dried plasma spots (DPSs as well as for the quantitation of whole blood 5-methyltetrahydrofolic acid in DBSs. We focused on two diagnostically conclusive parameters exhibited by the plasma and whole blood 5-methyltetrahydrofolic acid levels that reflect both temporary and long-term folate status. The method is performed using the [2H4]-labeled isotopologue of the vitamin as the internal standard, and three steps are required for the extraction procedure. Elution of the punched out matrix spots was performed using stabilization buffer including Triton X-100 in a standardized ultrasonication treatment followed by enzymatic digestion (whole blood only and solid-phase extraction with SAX cartridges. This method is sensitive enough to quantify 27 nmol/L whole blood 5-methyltetrahydrofolic acid in DBSs and 6.3 and 4.4 nmol/L plasma 5-methyltetrahydrofolic acid in DBSs and DPSs, respectively. The unprecedented accurate quantification of plasma 5-methyltetrahydrofolic acid in DBSs was achieved by thermal treatment prior to ultrasonication, inhibiting plasma conjugase activity. Mass screenings are more feasible and easier to facilitate for this method in terms of sample collection and storage compared with conventional clinical sampling for the assessment of folate status.

  12. THE USE OF DRIED BLOOD SPOTS AND A MODIFIED PARTICLE ...

    African Journals Online (AJOL)

    The use of dried blood spots (DBS) for the testing of HIV-1 antibodies has been described since 1988.' Numerous studies have proved that this method of prevalence testing has high sensitivity and specificity and is easy to use, as well as having the advantage of being less expensive than conventional tests on whole blood.

  13. A Novel, Nondestructive, Dried Blood Spot-Based Hematocrit Prediction Method Using Noncontact Diffuse Reflectance Spectroscopy

    NARCIS (Netherlands)

    Capiau, S.; Wilk, L.S.; Aalders, M.C.G.; Stove, C.P.

    2016-01-01

    Dried blood spot (DBS) sampling is recognized as a valuable alternative sampling strategy both in research and in clinical routine. Although many advantages are associated with DBS sampling, its more widespread use is hampered by several issues, of which the hematocrit effect on DBS-based

  14. Retinol analysis in dried blood spots by HPLC.

    Science.gov (United States)

    Craft, N E; Haitema, T; Brindle, L K; Yamini, S; Humphrey, J H; West, K P

    2000-04-01

    There are many advantages to measuring vitamin A in dried blood spots (DBS) from a finger prick as compared to plasma collected by venipuncture. The advantages include easier collection, transport and storage; accessibility to younger and more remote populations; and decreased risk of disease transmission. We describe a method for the extraction of retinol from DBS for analysis by HPLC and initial comparison to plasma retinol. The effects of various buffers, detergents, antioxidants and chelators were evaluated to establish the most effective approach to elute the retinol: retinol binding protein (holo-RBP) complex from the blood collection cards. The process involves ultrasonic agitation to elute holo-RBP into a phosphate buffer containing an antioxidant and metal chelator. The holo-RBP complex was denatured by the addition of ethanol containing additional antioxidants permitting the extraction of free retinol into hexane. Following solvent evaporation, the extract was dissolved in methanol for HPLC analysis. The initial measured retinol levels in freshly collected DBS declined for 6-10 d whether stored at 25, 4 or -20 degrees C, but remained consistent thereafter (homeostatic). By incorporating a "recovery/volume adjustment" factor, measured retinol values in homeostatic DBS were adjusted to the equivalent of plasma retinol. For 17 normal adults, the correlation coefficient was 0.90 between plasma retinol and adjusted DBS retinol in samples that had been stored at -70 degrees C for < 9 mo. The use of this new sample matrix for vitamin A assessment will allow access to previously unavailable populations.

  15. Manual versus automated blood sampling

    DEFF Research Database (Denmark)

    Teilmann, A C; Kalliokoski, Otto; Sørensen, Dorte B

    2014-01-01

    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......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...

  16. Dried blood spot analysis : facing new challenges

    NARCIS (Netherlands)

    Koster, Remco A; Touw, Daan J; Alffenaar, Jan-willem C

    2015-01-01

    Over the last decade, DBS analysis has gained popularity for TDM because it’s a patient friendly sampling proce- dure [1-4]. Additional advantages are prolonged sample stability, lower risk of infections and transportation at ambient temperature [1-3,5]. These advantages may fa- cilitate

  17. Validation and Application of a Dried Blood Spot Ceftriaxone Assay.

    Science.gov (United States)

    Page-Sharp, Madhu; Nunn, Troy; Salman, Sam; Moore, Brioni R; Batty, Kevin T; Davis, Timothy M E; Manning, Laurens

    2015-10-05

    Dried blood spot (DBS) antibiotic assays can facilitate pharmacokinetic/pharmacodynamic (PK/PD) studies in situations where venous blood sampling is logistically and/or ethically problematic. In this study, we aimed to develop, validate, and apply a DBS ceftriaxone assay. A liquid chromatography-tandem mass spectroscopy (LC-MS/MS) DBS ceftriaxone assay was assessed for matrix effects, process efficiency, recovery, variability, and limits of quantification (LOQ) and detection (LOD). The effects of hematocrit, protein binding, red cell partitioning, and chad positioning were evaluated, and thermal stability was assessed. Plasma, DBS, and cell pellet ceftriaxone concentrations in 10 healthy adults were compared, and plasma concentration-time profiles of DBS and plasma ceftriaxone were incorporated into population PK models. The LOQ and LOD for ceftriaxone in DBS were 0.14 mg/liter and 0.05 mg/liter, respectively. Adjusting for hematocrit, red cell partitioning, and relative recovery, DBS-predicted plasma concentrations were comparable to measured plasma concentrations (r > 0.95, P ceftriaxone retained >95% initial concentrations in DBS for 14 h, 35 h, 30 days, 21 weeks, and >11 months, respectively. The present DBS ceftriaxone assay is robust and can be used as a surrogate for plasma concentrations to provide valid PK and PK/PD data in a variety of clinical situations, including in studies of young children and of those in remote or resource-poor settings. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  18. Therapeutic drug monitoring of everolimus using the dried blood spot method in combination with liquid chromatography-mass spectrometry.

    Science.gov (United States)

    van der Heijden, J; de Beer, Y; Hoogtanders, K; Christiaans, M; de Jong, G J; Neef, C; Stolk, L

    2009-11-01

    An assay of everolimus based on finger prick sampling and consecutive application as a blood spot on sampling paper has been developed. We explored several methods [K. Hoogtanders, J. van der Heijden, M. Christiaans, P. Edelbroek, J. van Hooff, L. Stolk, J. Pharm. Biomed. Anal. 44 (2006) 658-664; A. Allanson, M. Cotton, J. Tettey, et al., J. Pharm. Biomed. Anal. 44 (2007) 963-969] and developed a new method, namely the impregnation of sampling paper with a solution of plasma-protein, formic acid and ammonium acetate, in combination with the extraction of the blood spot by filter filtration. This kind of sample preparation provides new possibilities for blood spot sampling especially if analytes are adsorbed to the paper. The dried blood spot was analysed using the HPLC-electrospray-tandem mass spectrometry method, with 32-desmethoxyrapamycin as the internal standard. The working range of our study was 2-30 microg/l. Within this range, intra-and inter-assay variability for precision and accuracy was spot samples proved stable for 3 days at 60 degrees C and for 32 days at 4 degrees C. Everolimus concentrations of one stable out-patient were compared after both blood spot sampling and conventional venous sampling on various occasions. Results indicate that this new method is promising for therapeutic drug monitoring in stable renal transplant patients.

  19. School based screening for hypothyroidism in Down's syndrome by dried blood spot TSH measurement

    OpenAIRE

    Noble, S; Leyland, K; Findlay, C; Clark, C; Redfern, J; Mackenzie, J; Girdwood, R; Donaldson, M

    2000-01-01

    OBJECTIVE—To determine the feasibility of annual hypothyroid screening of children with Down's syndrome by measuring thyroid stimulating hormone (TSH) on dried blood spots at school, and to describe the outcome in positive children.
DESIGN—Establishment of a register of school children with Down's syndrome, and procedures for obtaining permission from parents, annual capillary blood samples, TSH measurement, and clinical assessment of children with TSH values > 10 mU/litr...

  20. Plasma androgen concentrations in initial samples from spotted ...

    African Journals Online (AJOL)

    1990-01-31

    Jan 31, 1990 ... 1991,26(1). Plasma androgen concentrations in initial samples from spotted hyaenas immobilized with Zoletil (CI-744) reflect hormonal status estimated by GnRH challenge and immobilization stress response. A.S. van Jaarsveld * and J.D. Skinner. Mammal Research Institute, University of Pretoria, Pretoria ...

  1. Estimating population salt intake in India using spot urine samples.

    Science.gov (United States)

    Petersen, Kristina S; Johnson, Claire; Mohan, Sailesh; Rogers, Kris; Shivashankar, Roopa; Thout, Sudhir Raj; Gupta, Priti; He, Feng J; MacGregor, Graham A; Webster, Jacqui; Santos, Joseph Alvin; Krishnan, Anand; Maulik, Pallab K; Reddy, K Srinath; Gupta, Ruby; Prabhakaran, Dorairaj; Neal, Bruce

    2017-11-01

    To compare estimates of mean population salt intake in North and South India derived from spot urine samples versus 24-h urine collections. In a cross-sectional survey, participants were sampled from slum, urban and rural communities in North and in South India. Participants provided 24-h urine collections, and random morning spot urine samples. Salt intake was estimated from the spot urine samples using a series of established estimating equations. Salt intake data from the 24-h urine collections and spot urine equations were weighted to provide estimates of salt intake for Delhi and Haryana, and Andhra Pradesh. A total of 957 individuals provided a complete 24-h urine collection and a spot urine sample. Weighted mean salt intake based on the 24-h urine collection, was 8.59 (95% confidence interval 7.73-9.45) and 9.46 g/day (8.95-9.96) in Delhi and Haryana, and Andhra Pradesh, respectively. Corresponding estimates based on the Tanaka equation [9.04 (8.63-9.45) and 9.79 g/day (9.62-9.96) for Delhi and Haryana, and Andhra Pradesh, respectively], the Mage equation [8.80 (7.67-9.94) and 10.19 g/day (95% CI 9.59-10.79)], the INTERSALT equation [7.99 (7.61-8.37) and 8.64 g/day (8.04-9.23)] and the INTERSALT equation with potassium [8.13 (7.74-8.52) and 8.81 g/day (8.16-9.46)] were all within 1 g/day of the estimate based upon 24-h collections. For the Toft equation, estimates were 1-2 g/day higher [9.94 (9.24-10.64) and 10.69 g/day (9.44-11.93)] and for the Kawasaki equation they were 3-4 g/day higher [12.14 (11.30-12.97) and 13.64 g/day (13.15-14.12)]. In urban and rural areas in North and South India, most spot urine-based equations provided reasonable estimates of mean population salt intake. Equations that did not provide good estimates may have failed because specimen collection was not aligned with the original method.

  2. Method for the Determination of Iodide in Dried Blood Spots from Newborns by High Performance Liquid Chromatography Tandem Mass Spectrometry.

    Science.gov (United States)

    Kim, Un-Jung; Kannan, Kurunthachalam

    2018-03-06

    Dried blood spots (DBS), collected for newborn screening programs in the United States, have been used to screen for congenital metabolic diseases in newborns for over 50 years. DBS provide an easy and inexpensive way to collect and store peripheral blood specimens and present an excellent resource for studies on the assessment of chemical exposures in newborns. In this study, a selective and sensitive method was developed for the analysis of iodide in DBS by high performance liquid chromatography electrospray tandem mass spectrometry. Accuracy, inter- and intraday precision, matrix effects, and detection limits of the method were determined. Further validation of the method was accomplished by concurrent analysis of whole blood and fortified blood spotted on a Whatman 903 filter card. A significant positive correlation was found between measured concentrations of iodide in venous whole blood and the same blood spotted as DBS. The method limit of detection was 0.15 ng/mL iodide. The method was further validated by the analysis of a whole blood sample certified for iodide levels (proficiency testing sample) by spotting on a filter card. Twenty DBS samples collected from newborns in New York State were analyzed to demonstrate the applicability of the method. The measured concentrations of iodide in whole blood of newborns from New York State ranged between

  3. A simple method to quantitate IP-10 in dried blood and plasma spots.

    Directory of Open Access Journals (Sweden)

    Martine G Aabye

    Full Text Available BACKGROUND: Antigen specific release of IP-10 is an established marker for infection with M.tuberculosis. Compared to IFN-γ, IP-10 is released in 100-fold higher concentrations enabling the development of novel assays for detection. Dried blood spots are a convenient sample for high throughput newborn screening. AIM: To develop a robust and sensitive ELISA-based assay for IP-10 detection in plasma, dried blood spots (DBS and dried plasma spots (DPS; to validate the ELISA in clinically relevant samples; and to assess the performance of the assay for detection of Cytomegalovirus (CMV and M.tuberculosis specific immune responses. METHOD: We raised mice and rat monoclonal antibodies against human IP-10 and developed an ELISA. The assay was validated and applied to the detection of CMV and M.tuberculosis specific responses in 18 patients with immune reactivity towards M.tuberculosis and 32 healthy controls of which 22 had immune reactivity towards CMV and none towards M.tuberculosis. We compared the performance of this new assay to IFN-γ. RESULTS: The ELISA was reliable for IP-10 detection in both plasma and filter paper samples. The linear range of the ELISA was 2.5-600 pg/ml. IFN-γ was not readily detectable in DPS samples. IP-10 was stabile in filter paper samples for at least 4 weeks at 37 °C. The correlation between IP-10 detected in plasma, DPS and DBS samples was excellent (r(2>0.97. CONCLUSIONS: This newly developed assay is reliable for IP-10 quantification in plasma, DBS and DPS samples from antigen stimulated and non-stimulated whole blood. The filter paper assays enable easy sample acquisition and transport at ambient temperature e.g. via the postal system. The system can potentially simplify diagnostic assays for M.tuberculosis and CMV infection.

  4. Improving FoRe: A New Inlet Design for Filtering Samples through Individual Microarray Spots.

    Science.gov (United States)

    de Lange, Victoria; Habegger, Marco; Schmidt, Marco; Vörös, János

    2017-03-24

    In this publication we present an improvement to our previously introduced vertical flow microarray, the FoRe array, which capitalizes on the fusion of immunofiltration and densely packed micron test sites. Filtering samples through individual microarray spots allows us to rapidly analyze dilute samples with high-throughput and high signal-to-noise. Unlike other flowthrough microarrays, in the FoRe design samples are injected into micron channels and sequentially exposed to different targets. This arrangement makes it possible to increase the sensitivity of the microarray by simply increasing the sample volume or to rapidly reconcentrate samples after preprocessing steps dilute the analyte. Here we present a new inlet system which allows us to increase the analyzed sample volume without compromising the micron spot size and dense layout. We combined this with a model assay to demonstrate that the device is sensitive to the amount of antigen, and as a result, sample volume directly correlates to sensitivity. We introduced a simple technique for analysis of blood, which previously clogged the nanometer-sized pores, requiring only microliter volumes expected from an infant heel prick. A drop of blood is mixed with buffer to separate the plasma before reconcentrating the sample on the microarray spot. We demonstrated the success of this procedure by spiking TNF-α into blood and achieved a limit of detection of 18 pM. Compared to traditional protein microarrays, the FoRe array is still inexpensive, customizable, and simple to use, and thanks to these improvements has a broad range of applications from small animal studies to environmental monitoring.

  5. Neonatal blood gas sampling methods

    African Journals Online (AJOL)

    adequate collateral circulation before radial arterial puncture may not be a reliable predictor of subsequent risk of vascular injury.75. Conclusion and recommendations. Indwelling arterial catheters remain a practical, reliable and accurate method of neonatal blood gas sampling, provided they are inserted and maintained ...

  6. Carbamazepine, lamotrigine, levetiracetam and valproic acid in dried blood spots with liquid chromatography tandem mass spectrometry; method development and validation.

    Science.gov (United States)

    Linder, Camilla; Hansson, Anna; Sadek, Sara; Gustafsson, Lars L; Pohanka, Anton

    2018-01-01

    Monitoring of antiepileptic drugs in children with epilepsy require multiple visits at a clinic for blood collection. Dried blood spot sampling is an alternative way of collection, performed at home by self-collection and can save time and costs for patients and family members. The aim was to develop and validate an LC-MS/MS dried blood spot method for carbamazepine, lamotrigine, levetiracetam and valproic acid with the requirements of using standard equipment and material in a routine laboratory setting. Whatman-903 filter paper was utilized, and discs were punched into a 96 well plate with an automated puncher and barcode reading. Extraction with methanol/water solution including internal standards on an orbital shaker was followed by a vacuum centrifuge step and reconstitution in mobile phase. Bioanalytical validation was performed according to guidelines from European Medicines Agency and additional dried blood spot specific validation. Calibration curves of the four included drugs had R 2 values ≥0.994. Therapeutic relevant concentrations were well within measuring ranges. Within and -between run precision had %CV:s of 2.9-10.5%. Accuracy (%bias) was between -16.5% (lower limit of quantification) to +7.4%. Blood spots in a volume range of 15-50μL with hematocrit in expected ranges for this patient group were within precision and accuracy limits. To test the method, concentrations from dried blood spot venous and capillary patient samples (n=50) were compared with plasma concentrations. Good correlations for all four drugs with R 2 of >0.92 was shown. In summary, a fast method for dried blood spots based on a 96 well format was developed for four commonly prescribed antiepileptic drugs. This validated method with traceability in sample preparation by bar code reading makes it suitable for the clinical laboratory. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Design of an online spot air sampling system

    International Nuclear Information System (INIS)

    Rath, D.P.; Govalkar, Atul; Yadav, S.A.; Mukherjee, Govinda; Murali, S.; Babu, D.A.R.; Prasad, R.S.

    2012-01-01

    In all the alpha handling facilities spot air sampling is essential while clearing a lab consequent to week end ventilation shut down or subsequent to completion of work/operations goes inside the facility. The sampling head is designed for 5 micron particles. Following pints are noticed over a period of such operational activities: Health Physicist goes inside a lab with a half face mask whose protection factor is 10, thereby useful for lab air activity up to ten DACs. Many times HP goes inside the lab when activity is more which is highly unsafe. On many occasions simultaneously a number of samples have to be taken by HP from a large number of laboratories inside the facility or for any special job where continuous radiological protection is required. Based on this it is proposed to design a sampling system which will overcome the above limitations. The design of the sampling head has been carried out which is for 5 micron particle size. The pump which is available in the existing facilities can be utilized. An innovative way of counting for large number of samples is fabricated in the RHC wing, RMD which can count ten samples at a time. Removal and counting of the sample may be carried out in a similar sampling carousel which is being used in RHC Unit, Radiometallurgy wing successfully with a little modification. In the proposed system, three samples can be operated in-line such that health physics intervention during the active operation would be minimum and during alarm situations (i.e. on the DAC level) proper protective equipment shall be advised by health physicist or he may suggest any other protective action. This type of online monitors will help in establishing the airborne activity inside the lab where special jobs are being carried out which will provide maximum protection to the lab personnel as well as to the health physicist who supervises the entire operation

  8. An automated immunoradiometric assay of thyrotrophin (TSH) in dried blood filter paper spots

    International Nuclear Information System (INIS)

    John, R.; Woodhead, J.S.

    1982-01-01

    An immunoradiometric two-site assay for thyrotrophin (TSH) in dried blood filter paper spots is described. The assay is automated by means of the Kemtek 3000 automated immunoassay system. The technique uses a 6.0 mm disc punched from the dried blood samples collected as part of the screening programme for phenylketonuria. The method is sensitive and precise, and results correlate well with those obtained in TSH assays of serum samples. The procedure is rapid, results being available within 24 h of receipt of samples. Of 25204 specimens so far screened by this assay, 99.9% have TSH levels less than 15 mU/l. One false positive result has been obtained and six confirmed cases of neonatal hypothyroidism detected, giving a prevalence of 1 in 4200. (Auth.)

  9. Automated immunoradiometric assay of thyrotrophin (TSH) in dried blood filter paper spots

    Energy Technology Data Exchange (ETDEWEB)

    John, R.; Woodhead, J.S. (Welsh National School of Medicine, Cardiff (UK))

    1982-11-10

    An immunoradiometric two-site assay for thyrotrophin (TSH) in dried blood filter paper spots is described. The assay is automated by means of the Kemtek 3000 automated immunoassay system. The technique uses a 6.0 mm disc punched from the dried blood samples collected as part of the screening programme for phenylketonuria. The method is sensitive and precise, and results correlate well with those obtained in TSH assays of serum samples. The procedure is rapid, results being available within 24 h of receipt of samples. Of 25204 specimens so far screened by this assay, 99.9% have TSH levels less than 15 mU/l. One false positive result has been obtained and six confirmed cases of neonatal hypothyroidism detected, giving a prevalence of 1 in 4200.

  10. The Influence of Education on Public Trust and Consent Preferences With Residual Newborn Screening Dried Blood spots.

    Science.gov (United States)

    Rothwell, Erin; Wong, Bob; Anderson, Rebecca A; Botkin, Jeffrey R

    2016-07-01

    The objectives of this study were to evaluate the impact of educational interventions during prenatal care on public trust for newborn screening and consent preferences for the retention and use of leftover newborn screening dried blood spots. Women who were 30 to 36 weeks pregnant were recruited, and outcomes were measured by telephone survey 2 to 4 weeks postpartum (n = 901). Approximately 40% of the sample chose the opt-out approach but those who watched educational interventions delivered during prenatal care were significantly associated with higher levels of trust and support for an opt-out consent approach. Providing education during prenatal care about newborn screening and the storage and use of leftover dried blood spots along with brochure-based education provided in the hospital when the baby is born is associated with improved trust for the program and support for research with the leftover blood spots. © The Author(s) 2016.

  11. Fully automated drug screening of dried blood spots using online LC-MS/MS analysis

    Directory of Open Access Journals (Sweden)

    Stefan Gaugler

    2018-01-01

    Full Text Available A new and fully automated workflow for the cost effective drug screening of large populations based on the dried blood spot (DBS technology was introduced in this study. DBS were prepared by spotting 15 μL of whole blood, previously spiked with alprazolam, amphetamine, cocaine, codeine, diazepam, fentanyl, lysergic acid diethylamide (LSD, 3,4-methylenedioxymethamphet-amine (MDMA, methadone, methamphetamine, morphine and oxycodone onto filter paper cards. The dried spots were scanned, spiked with deuterated standards and directly extracted. The extract was transferred online to an analytical LC column and then to the electrospray ionization tandem mass spectrometry system. All drugs were quantified at their cut-off level and good precision and correlation within the calibration range was obtained. The method was finally applied to DBS samples from two patients with back pain and codeine and oxycodone could be identified and quantified accurately below the level of misuse of 89.6 ng/mL and 39.6 ng/mL respectively.

  12. Validation study of the Tanaka and Kawasaki equations to estimate the daily sodium excretion by a spot urine sample.

    Science.gov (United States)

    Mill, José Geraldo; Rodrigues, Sérgio Lamêgo; Baldo, Marcelo Perim; Malta, Deborah Carvalho; Szwarcwald, Celia Landmann

    2015-12-01

    To validate Tanaka and Kawasaki's formulas to calculate the salt intake by the sodium/creatinine ratio in spot of urine. Two hundred and seventy two adults (20 - 69 years old; 52.6% women) with 24 h urine collection and two urinary spots collected on the same day (while fasting - spot 1 - or not fasting - spot 2). Anthropometry, blood pressure and fasting blood were measured on the same day. The analysis of agreement between salt consumption measured in the 24 h urine test and urinary spots were determined by the Pearson's correlation (r) and the Bland & Altman method. The mean salt consumption measured by the 24 h sodium excretion was 10.4 ± 5.3 g/day. The correlation between the measured 24 h sodium excretion and the estimation based on spots 1 and 2, respectively, was only moderated according to Tanaka (r = 0.51 and r = 0.55; p salt consumption by Tanaka to increasing salt consumption and conversely, an overestimation of consumption by the Kawasaki formula. The estimation of salt consumption (difference between measured and calculated salt consumption lower than 1 g/day) was adequate only when the consumption was between 9 - 12 g/day (Tanaka) and 12 - 18 g/day (Kawasaki). Spot urine sampling is adequate to estimate salt consumption only among individuals with an actual consumption near the population mean.

  13. Dried blood spot validation of five immunosuppressants, without hematocrit correction, on two LC-MS/MS systems

    NARCIS (Netherlands)

    Koster, Remco A; Veenhof, Herman; Botma, Rixt; Hoekstra, Alle Tjipke; Berger, Stefan P; Bakker, Stephan JL; Alffenaar, Jan-Willem C; Touw, Daan J

    2017-01-01

    AIM: Hematocrit (Ht) effects remain a challenge in dried blood spot (DBS) sampling. The aim was to develop an immunosuppressant DBS assay on two LC-MS/MS systems covering a clinically relevant Ht range without Ht correction. RESULTS: The method was partially validated for tacrolimus, sirolimus,

  14. Percutaneous umbilical cord blood sampling - slideshow

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/presentations/100196.htm Percutaneous umbilical cord blood sampling - series—Normal anatomy To use the ... or blood disorder, your doctor may recommend percutaneous umbilical cord blood sampling (PUBS), which is performed at 18 ...

  15. Silica Coated Paper Substrate for Paper-Spray Analysis of Therapeutic Drugs in Dried Blood Spots

    Science.gov (United States)

    Zhang, Zhiping; Xu, Wei; Manicke, Nicholas E.; Cooks, R. Graham; Ouyang, Zheng

    2011-01-01

    Paper spray is a newly developed ambient ionization method that has been applied for direct qualitative and quantitative analysis of biological samples. The properties of the paper substrate and spray solution have a significant impact on the release of chemical compounds from complex sample matrices, the diffusion of the analytes through the substrate, and the formation of ions for mass spectrometry analysis. In this study, a commercially available silica-coated paper was explored in an attempt to improve the analysis of therapeutic drugs in dried blood spots (DBS). The dichloromethane/isopropanol solvent has been identified as an optimal spray solvent for the analysis. The comparison was made with paper spray using chromatography paper as substrate with methanol/water as solvent for the analysis of verapamil, citalopram, amitriptyline, lidocaine and sunitinib in dried blood spots. It has been demonstrated the efficiency of recovery of the analytes was notably improved with the silica coated paper and the limit of quantitation (LOQ) for the drug analysis was 0.1 ng mL−1 using a commercial triple quadrupole mass spectrometer. The use of silica paper substrate also resulted in a sensitivity improvement of 5-50 fold in comparison with chromatography papers, including the Whatmann ET31 paper used for blood card. Analysis using a handheld miniature mass spectrometer Mini 11 gave LOQs of 10~20 ng mL−1 for the tested drugs, which is sufficient to cover the therapeutic ranges of these drugs. PMID:22145627

  16. Use of Dried Blood Spots to Elucidate Full-Length Transmitted/Founder HIV-1 Genomes

    Directory of Open Access Journals (Sweden)

    Jesus F. Salazar-Gonzalez

    2016-07-01

    Full Text Available Background: Identification of HIV-1 genomes responsible for establishing clinical infection in newly infected individuals is fundamental to prevention and pathogenesis research. Processing, storage, and transportation of the clinical samples required to perform these virologic assays in resource-limited settings requires challenging venipuncture and cold chain logistics. Here, we validate the use of dried-blood spots (DBS as a simple and convenient alternative to collecting and storing frozen plasma. Methods: We performed parallel nucleic acid extraction, single genome amplification (SGA, next generation sequencing (NGS, and phylogenetic analyses on plasma and DBS. Results: We demonstrated the capacity to extract viral RNA from DBS and perform SGA to infer the complete nucleotide sequence of the transmitted/founder (TF HIV-1 envelope gene and full-length genome in two acutely infected individuals. Using both SGA and NGS methodologies, we showed that sequences generated from DBS and plasma display comparable phylogenetic patterns in both acute and chronic infection. SGA was successful on samples with a range of plasma viremia, including samples as low as 1,700 copies/ml and an estimated ~50 viral copies per blood spot. Further, we demonstrated reproducible efficiency in gp160 env sequencing in DBS stored at ambient temperature for up to three weeks or at -20ºC for up to five months. Conclusions: These findings support the use of DBS as a practical and cost-effective alternative to frozen plasma for clinical trials and translational research conducted in resource-limited settings.

  17. Hyperspectral imaging and multivariate analysis in the dried blood spots investigations

    Science.gov (United States)

    Majda, Alicja; Wietecha-Posłuszny, Renata; Mendys, Agata; Wójtowicz, Anna; Łydżba-Kopczyńska, Barbara

    2018-04-01

    The aim of this study was to apply a new methodology using the combination of the hyperspectral imaging and the dry blood spot (DBS) collecting. Application of the hyperspectral imaging is fast and non-destructive. DBS method offers the advantage also on the micro-invasive blood collecting and low volume of required sample. During experimental step, the reflected light was recorded by two hyperspectral systems. The collection of 776 spectral bands in the VIS-NIR range (400-1000 nm) and 256 spectral bands in the SWIR range (970-2500 nm) was applied. Pixel has the size of 8 × 8 and 30 × 30 µm for VIS-NIR and SWIR camera, respectively. The obtained data in the form of hyperspectral cubes were treated with chemometric methods, i.e., minimum noise fraction and principal component analysis. It has been shown that the application of these methods on this type of data, by analyzing the scatter plots, allows a rapid analysis of the homogeneity of DBS, and the selection of representative areas for further analysis. It also gives the possibility of tracking the dynamics of changes occurring in biological traces applied on the surface. For the analyzed 28 blood samples, described method allowed to distinguish those blood stains because of time of apply.

  18. Evaluation of DNA extraction methods for the detection of Cytomegalovirus in dried blood spots

    Science.gov (United States)

    Koontz, D.; Baecher, K.; Amin, M.; Nikolova, S.; Gallagher, M.; Dollard, S.

    2015-01-01

    Background Dried blood spots (DBS) are collected universally from newborns and may be valuable for the diagnosis of congenital Cytomegalovirus (CMV) infection. The reported analytical sensitivity for DBS testing compared to urine or saliva varies greatly across CMV studies. The purpose of this study was to directly compare the performance of various DNA extraction methods for identification of CMV in DBS including those used most often in CMV studies. Study design Whatman® Grade 903 filter paper cards were spotted with blood samples from 25 organ transplant recipients who had confirmed CMV viremia. Six DNA extraction methods were compared for relative yield of viral and cellular DNA: 2 manual solution-based methods (Gentra Puregene, thermal shock), 2 manual silica column-based methods (QIAamp DNA Mini, QIAamp DNA Investigator), and 2 automated methods (M48 MagAttract Mini, QIAcube Investigator). DBS extractions were performed in triplicate followed by real-time quantitative PCR (qPCR). Results For extraction of both viral and cellular DNA, two methods (QIAamp DNA Investigator and thermal shock) consistently gave the highest yields, and two methods (M48 MagAttract Mini and QIAamp DNA Mini) consistently gave the lowest yields. There was an average 3-fold difference in DNA yield between the highest and lowest yield methods. Conclusion The choice of DNA extraction method is a major factor in the ability to detect low levels of CMV in DBS and can largely account for the wide range of DBS sensitivities reported in studies to date. PMID:25866346

  19. Common criteria among States for storage and use of dried blood spot specimens after newborn screening

    Directory of Open Access Journals (Sweden)

    Carlo Petrini

    2012-06-01

    Full Text Available Biological samples collected in biobanks are a resource with significant research potential. The Italian Joint Group cNB - cNBBSV (National committee of Bioethics - National committee for Biosecurity, Biotechnologies and Life Sciences published a document reporting recommendations on storage and use of dried blood spot (DBS and on the development of a National Network of Regional Newborn Screening Repositories for collection of residual DBS. Several ethical questions (about consent, possible use of genetic information, unanticipated possible usages for research purposes rise from residual newborn screening specimens collections. Moreover, legal and ethical controversies are accentuated by the conflicts between the interests of sample donors, biobank holders, researchers and the public. To overcome these difficulties the identification of a few criteria for storage and research usage of DBS is crucial.

  20. A novel polymerase chain reaction method for detection of human immunodeficiency virus in dried blood spots on filter paper.

    Science.gov (United States)

    Yourno, J; Conroy, J

    1992-11-01

    A method for detection of proviral human immunodeficiency virus DNA in dried blood spots on filter paper by direct polymerase chain reaction (PCR) has been developed. To develop the method, a standard system was used which was prepared from cells each containing a single integrated provirus and titrated with normal donor blood. This rapid procedure provides virtually quantitative yields of nuclear DNA and exploits most of the standard methodology described for blood specimens. A nested PCR using SK38-SK39 gag as the internal primer pair was also designed; this PCR detected a single copy of provirus per filter at near theoretical frequency with SK19 probe. The utility of the procedure was demonstrated with clinical specimens. Blood spot filters from human immunodeficiency virus-infected and uninfected individuals were readily and unequivocally discriminated. The method is designed for ultimate use with large (1.5-ml) sample preparation tubes that are compatible as PCR tubes with thermal cyclers. This will permit convenient, direct single-tube PCR of dried blood specimens on filters. It should be adaptable to analysis of dried blood spots for a variety of infectious or genetic diseases.

  1. Whole genome sequencing of Plasmodium falciparum from dried blood spots using selective whole genome amplification.

    Science.gov (United States)

    Oyola, Samuel O; Ariani, Cristina V; Hamilton, William L; Kekre, Mihir; Amenga-Etego, Lucas N; Ghansah, Anita; Rutledge, Gavin G; Redmond, Seth; Manske, Magnus; Jyothi, Dushyanth; Jacob, Chris G; Otto, Thomas D; Rockett, Kirk; Newbold, Chris I; Berriman, Matthew; Kwiatkowski, Dominic P

    2016-12-20

    Translating genomic technologies into healthcare applications for the malaria parasite Plasmodium falciparum has been limited by the technical and logistical difficulties of obtaining high quality clinical samples from the field. Sampling by dried blood spot (DBS) finger-pricks can be performed safely and efficiently with minimal resource and storage requirements compared with venous blood (VB). Here, the use of selective whole genome amplification (sWGA) to sequence the P. falciparum genome from clinical DBS samples was evaluated, and the results compared with current methods that use leucodepleted VB. Parasite DNA with high (>95%) human DNA contamination was selectively amplified by Phi29 polymerase using short oligonucleotide probes of 8-12 mers as primers. These primers were selected on the basis of their differential frequency of binding the desired (P. falciparum DNA) and contaminating (human) genomes. Using sWGA method, clinical samples from 156 malaria patients, including 120 paired samples for head-to-head comparison of DBS and leucodepleted VB were sequenced. Greater than 18-fold enrichment of P. falciparum DNA was achieved from DBS extracts. The parasitaemia threshold to achieve >5× coverage for 50% of the genome was 0.03% (40 parasites per 200 white blood cells). Over 99% SNP concordance between VB and DBS samples was achieved after excluding missing calls. The sWGA methods described here provide a reliable and scalable way of generating P. falciparum genome sequence data from DBS samples. The current data indicate that it will be possible to get good quality sequence on most if not all drug resistance loci from the majority of symptomatic malaria patients. This technique overcomes a major limiting factor in P. falciparum genome sequencing from field samples, and paves the way for large-scale epidemiological applications.

  2. Blood parasites in Owls with conservation implications for the Spotted Owl (Strix occidentalis)

    Science.gov (United States)

    Ishak, H.D.; Dumbacher, J.P.; Anderson, N.L.; Keane, J.J.; Valkiunas, G.; Haig, S.M.; Tell, L.A.; Sehgal, R.N.M.

    2008-01-01

    The three subspecies of Spotted Owl (Northern, Strix occidentalis courina; California, S. o. occidentalis; and Mexican, S. o. lucida) are all threatened by habitat loss and range expansion of the Barred Owl (S. varia). An unaddressed threat is whether Barred Owls could be a source of novel strains of disease such as avian malaria (Plasmodium spp.) or other blood parasites potentially harmful for Spotted Owls. Although Barred Owls commonly harbor Plasmodium infections, these parasites have not been documented in the Spotted Owl. We screened 111 Spotted Owls, 44 Barred Owls, and 387 owls of nine other species for haemosporidian parasites (Leucocytozoon, Plasmodium, and Haemoproteus spp.). California Spotted Owls had the greatest number of simultaneous multi-species infections (44%). Additionally, sequencing results revealed that the Northern and California Spotted Owl subspecies together had the highest number of Leucocytozoon parasite lineages (n=17) and unique lineages (n=12). This high level of sequence diversity is significant because only one leucocytozoon species (L. danilewskyi) has been accepted as valid among all owls, suggesting that L. danilewskyi is a cryptic species. Furthermore, a Plasmodium parasite was documented in a Northern Spotted Owl for the first time. West Coast Barred Owls had a lower prevalence of infection (15%) when compared to sympatric Spotted Owls (S. o. caurina 52%, S. o. occidentalis 79%) and Barred Owls from the historic range (61%). Consequently, Barred Owls on the West Coast may have a competitive advantage over the potentially immune compromised Spotted Owls. ?? 2008 Ishak et al.

  3. External quality assessment of cytomegalovirus DNA detection on dried blood spots

    Directory of Open Access Journals (Sweden)

    Binda Sandro

    2008-01-01

    Full Text Available Abstract Background Testing for viral DNA in neonatal blood dried on paper (DBS has proved a valid means of diagnosing congenital CMV infection with both clinical and epidemiological relevance. To assess the quality of the detection of CMV-DNA on DBS in laboratories performing this test a proficiency panel consisting of nine samples with two blood spots on each filter paper was produced and distributed. Six samples were derived from whole blood, negative for CMV DNA and antibody, and spiked with cell-grown CMV Towne in various concentrations (7.3 × 102 – 9.6 × 105 copies/ml, one was a CMV positive clinical specimen (3.9 × 106 copies/ml, and two samples were CMV-negative whole blood. Results The 27 responding laboratories from 14 countries submitted 33 datasets obtained by means of conventional PCR (n = 5 or real-time PCR (n = 28 technologies. A correct positive result was reported in at least 91% of datasets in samples with a viral load of 8.8 × 104 copies/ml or higher. However only 59% and 12% identified the 9.4 × 103 and 7.3 × 102 copies/ml samples, respectively, correctly as positive. False positive results were reported by 9% of laboratories and in 11% of datasets. Conclusion These results indicate a clear need for improvement of methods as sensitivity and false-positivity still appear to be a major problem in a considerable number of laboratories.

  4. Application of Dried Blood Spots on Filter Paper for Detection of HIV ...

    African Journals Online (AJOL)

    Moreover the present technique of collecting blood for diagnosis is associated with increased risk of infection, loss and contamination of specimen. This study aimed at evaluating the use of blood spot dried on filter paper (under different storage conditions) for detection of HIV antibodies in patient\\'s serum as an alternative ...

  5. Evaluation of the human immunodeficiency virus type 1 and 2 antibodies detection in dried whole blood spots (dbs samples Pesquisa de anticorpos contra o vírus da imunodeficiência humana tipos 1 e 2 em amostras de sangue seco coletadas em papel filtro

    Directory of Open Access Journals (Sweden)

    Andréa Cauduro de Castro

    2008-06-01

    Full Text Available Human Immunodeficiency Vírus Type 1 and 2 antibodies detection was performed in 457 dried whole blood spots samples (S&S 903. Q-Preven HIV 1+2 was the screening test used. The results were compared with the gold standard serum tests by ELISA (Cobas Core e Axsym HIV1/2 gO and imunofluorescence was the definitive confirmatory test. The samples were obtained from the Hospital Nossa Senhora da Conceição in Porto Alegre, RS - Brazil, through whole blood transfer to filter paper card and sent to Caxias do Sul, RS - Brazil where the tests were performed. The dried whole blood spot stability was evaluated with two different panels. The first one was composed of five negative and five positive samples stored at room temperature, 4 ºC, -20 ºC and -70 ºC, while the second was composed of two negative and three positive samples stored at 37 ºC (humidity Foram realizados 457 testes para detectar anticorpos contra o Vírus da Imunodeficiência Humana tipos 1 e 2, em amostras de sangue total seco coletadas em papel filtro (S&S 903, com o teste de triagem Q-Preven HIV 1+2, comparando-se com os resultados dos testes de triagem no soro (Cobas Core e Axsym HIV1/2 gO, sendo a imunofluorescência indireta o teste confirmatório. As amostras foram obtidas no Hospital Conceição em Porto Alegre, pela transferência de sangue total para cartão de papel filtro e encaminhadas para Caxias do Sul para a realização dos testes. Foi analisada a estabilidade da amostra em papel filtro com a utilização de dois painéis: o primeiro com cinco amostras negativas e cinco positivas armazenadas por seis semanas à temperatura ambiente, 4 ºC, -20 ºC e -70 ºC; o segundo com duas negativas e três positivas armazenadas por seis semanas com avaliações semanais a 37 ºC (umidade <50%. Os resultados de todas as amostras testadas foram mantidos. A sensibilidade foi de 100%, a especificidade de 99,6%, o valor preditivo positivo de 99,5% e o valor preditivo negativo de 100

  6. Evaluation of the dried blood spot filter paper technology and five testing strategies of HIV-1 and HIV-2 infections in West Africa

    NARCIS (Netherlands)

    Sarge-Njie, Ramu; Schim van der Loeff, Maarten; Ceesay, Saihou; Cubitt, David; Sabally, Saihou; Corrah, Tumani; Whittle, Hilton

    2006-01-01

    Simple robust approaches are needed to monitor the prevalence and incidence of HIV in Africa. The aim of this study was to evaluate the use of dried blood spot (DBS) as an alternative to serum or plasma for sentinel surveillance. Paired DBS and blood samples were obtained from 200 patients attending

  7. Diagnosis of Morquio Syndrome in Dried Blood Spots Based on a New MRM-MS Assay.

    Directory of Open Access Journals (Sweden)

    Claudia Cozma

    Full Text Available Mucopolysaccharidosis IVA (MPS IVA; Morquio A disease is an autosomal recessive disease caused and characterized by a decreased activity of N-acetylgalactosamine-6-sulfate sulfatase (GALNS, resulting in accumulation of keratan sulfate and chondroitin-6-sulfate in tissues and secondary organ damage. Recently approved enzyme replacement therapy renders the easy and early identification of MPS IVA of out-most importance.We propose a completely new assay for the stable and reproducible detection of GALNS deficiency in dry blood spots (DBS. For the validation blood samples were taken from 59 healthy individuals and 24 randomly selected genetically confirmed MPS IVA patients. The material extracted from DBS was incubated with a 4-methylumbelliferyl-β-D-galactopyranoside-6-sulfate as a specific substrate. Final enzymatic product, 4-methylumbelliferone, obtained after adding exogenous beta-galactosidase, was quantified by LC/MRM-MS (liquid-chromatography/multiple-reaction-monitoring mass-spectrometry. 4-propyl-5-hydroxy-7-methyl-2h-chromen-2-one was used as internal standard, a compound with a similar molecular structure and fragmentation pattern in negative ion mode as 4-methylumbelliferone.The enzymatic assay yielded a positive and negative predictive value of 1.0 for genetically confirmed MPS IVA patients (GALNS activity of 0.35 ± 0.21 μmol/L/h and for controls with normal GALNS activity (23.1 ± 5.3 μmol/L /h. With present enzymatic conditions, the reaction yield in dried blood spots is at least 20 fold higher than any previously reported data with other assays.The present LC/MRM-MS based assay for MPS IVA diagnosis provides an easy, highly-standardized, accurate and innovative quantification of the enzymatic product in vitro and distinguishes perfectly between MPS IVA affected patients and normal controls. This technique will significantly simplify the early detection of MPS IVA patients.

  8. Diagnosis of Morquio Syndrome in Dried Blood Spots Based on a New MRM-MS Assay.

    Science.gov (United States)

    Cozma, Claudia; Eichler, Sabrina; Wittmann, Gyula; Flores Bonet, Alba; Kramp, Guido Johannes; Giese, Anne-Katrin; Rolfs, Arndt

    2015-01-01

    Mucopolysaccharidosis IVA (MPS IVA; Morquio A disease) is an autosomal recessive disease caused and characterized by a decreased activity of N-acetylgalactosamine-6-sulfate sulfatase (GALNS), resulting in accumulation of keratan sulfate and chondroitin-6-sulfate in tissues and secondary organ damage. Recently approved enzyme replacement therapy renders the easy and early identification of MPS IVA of out-most importance. We propose a completely new assay for the stable and reproducible detection of GALNS deficiency in dry blood spots (DBS). For the validation blood samples were taken from 59 healthy individuals and 24 randomly selected genetically confirmed MPS IVA patients. The material extracted from DBS was incubated with a 4-methylumbelliferyl-β-D-galactopyranoside-6-sulfate as a specific substrate. Final enzymatic product, 4-methylumbelliferone, obtained after adding exogenous beta-galactosidase, was quantified by LC/MRM-MS (liquid-chromatography/multiple-reaction-monitoring mass-spectrometry). 4-propyl-5-hydroxy-7-methyl-2h-chromen-2-one was used as internal standard, a compound with a similar molecular structure and fragmentation pattern in negative ion mode as 4-methylumbelliferone. The enzymatic assay yielded a positive and negative predictive value of 1.0 for genetically confirmed MPS IVA patients (GALNS activity of 0.35 ± 0.21 μmol/L/h) and for controls with normal GALNS activity (23.1 ± 5.3 μmol/L /h). With present enzymatic conditions, the reaction yield in dried blood spots is at least 20 fold higher than any previously reported data with other assays. The present LC/MRM-MS based assay for MPS IVA diagnosis provides an easy, highly-standardized, accurate and innovative quantification of the enzymatic product in vitro and distinguishes perfectly between MPS IVA affected patients and normal controls. This technique will significantly simplify the early detection of MPS IVA patients.

  9. High-Quality Exome Sequencing of Whole-Genome Amplified Neonatal Dried Blood Spot DNA.

    Directory of Open Access Journals (Sweden)

    Jesper Buchhave Poulsen

    Full Text Available Stored neonatal dried blood spot (DBS samples from neonatal screening programmes are a valuable diagnostic and research resource. Combined with information from national health registries they can be used in population-based studies of genetic diseases. DNA extracted from neonatal DBSs can be amplified to obtain micrograms of an otherwise limited resource, referred to as whole-genome amplified DNA (wgaDNA. Here we investigate the robustness of exome sequencing of wgaDNA of neonatal DBS samples. We conducted three pilot studies of seven, eight and seven subjects, respectively. For each subject we analysed a neonatal DBS sample and corresponding adult whole-blood (WB reference sample. Different DNA sample types were prepared for each of the subjects. Pilot 1: wgaDNA of 2x3.2mm neonatal DBSs (DBS_2x3.2 and raw DNA extract of the WB reference sample (WB_ref. Pilot 2: DBS_2x3.2, WB_ref and a WB_ref replica sharing DNA extract with the WB_ref sample. Pilot 3: DBS_2x3.2, WB_ref, wgaDNA of 2x1.6 mm neonatal DBSs and wgaDNA of the WB reference sample. Following sequencing and data analysis, we compared pairwise variant calls to obtain a measure of similarity--the concordance rate. Concordance rates were slightly lower when comparing DBS vs WB sample types than for any two WB sample types of the same subject before filtering of the variant calls. The overall concordance rates were dependent on the variant type, with SNPs performing best. Post-filtering, the comparisons of DBS vs WB and WB vs WB sample types yielded similar concordance rates, with values close to 100%. WgaDNA of neonatal DBS samples performs with great accuracy and efficiency in exome sequencing. The wgaDNA performed similarly to matched high-quality reference--whole-blood DNA--based on concordance rates calculated from variant calls. No differences were observed substituting 2x3.2 with 2x1.6 mm discs, allowing for additional reduction of sample material in future projects.

  10. Rapid assays of clozapine and its metabolites in dried blood spots by liquid chromatography and microextraction by packed sorbent procedure.

    Science.gov (United States)

    Saracino, Maria Addolorata; Lazzara, Giuseppe; Prugnoli, Benedetta; Raggi, Maria Augusta

    2011-04-22

    A novel analytical approach has been developed for the determination of clozapine and its metabolites in dried blood spots on filter paper, using a chromatographic method coupled with a microextraction by packed sorbent procedure. The analytes were separated on a RP-C18 column using a mobile phase composed of 20% methanol, 16% acetonitrile and 64% aqueous phosphate buffer. Coulometric detection was used, setting the guard cell at +0.050 V, the first analytical cell at -0.200 V and the second analytical cell at +0.500 V. Clozapine and its metabolites were extracted from dried blood spots with phosphate buffer and, then, a microextraction by packed sorbent procedure for the sample clean-up was implemented obtaining good extraction yields. The calibration curve was linear over the 2.5-1000 ng mL(-1) blood concentration ranges for all the analytes. The method validation gave satisfactory results in terms of sensitivity, precision, selectivity and accuracy. The analytical method was successfully applied to dried blood spots from several psychiatric patients for therapeutic drug monitoring purpose. Copyright © 2011 Elsevier B.V. All rights reserved.

  11. The development and validation of dried blood spots for external quality assurance of syphilis serology.

    Science.gov (United States)

    Smit, Pieter W; van der Vlis, Thomas; Mabey, David; Changalucha, John; Mngara, Julius; Clark, Benjamin D; Andreasen, Aura; Todd, Jim; Urassa, Mark; Zaba, Basia; Peeling, Rosanna W

    2013-02-26

    Syphilis causes up to 1,500,000 congenital syphilis cases annually. These could be prevented if all pregnant women were screened, and those with syphilis treated with a single dose of penicillin before 28 weeks gestation. In recent years, rapid point-of-care tests have allowed greater access to syphilis screening, especially in rural or remote areas, but the lack of quality assurance of rapid testing has been a concern. We determined the feasibility of using dried blood spots (DBS) as specimens for quality assurance of syphilis serological assays. We developed DBS extraction protocols for use with Treponema pallidum particle agglutination assay (TPPA), Treponema pallidum haemagglutination assay (TPHA) and an enzyme immunoassay (EIA) and compared the results with those using matching plasma samples from the same patient. Since DBS samples showed poor performance with TPHA and EIA (TPHA sensitivity was 50.5% (95% confidence interval: 39.9-61.2%) and EIA specificity was 50.4% (95% CI: 43.7-57.1%), only the DBS TPPA was used in the final evaluation. DBS TPPA showed an sensitivity of 95.5% (95% CI: 91.3-98.0%) and a specificity of 99.0% (95% CI: 98.1-99.5%) compared to TPPA using plasma samples as a reference. DBS samples can be recommended for use with TPPA, and may be of value for external quality assurance of point-of-care syphilis testing.

  12. Energy dispersive X-ray fluorescence spectrometry for the direct multi-element analysis of dried blood spots

    Science.gov (United States)

    Marguí, E.; Queralt, I.; García-Ruiz, E.; García-González, E.; Rello, L.; Resano, M.

    2018-01-01

    Home-based collection protocols for clinical specimens are actively pursued as a means of improving life quality of patients. In this sense, dried blood spots (DBS) are proposed as a non-invasive and even self-administered alternative to sampling whole venous blood. This contribution explores the potential of energy dispersive X-ray fluorescence spectrometry for the simultaneous and direct determination of some major (S, Cl, K, Na), minor (P, Fe) and trace (Ca, Cu, Zn) elements in blood, after its deposition onto clinical filter papers, thus giving rise to DBS. For quantification purposes the best strategy was to use matrix-matched blood samples of known analyte concentrations. The accuracy and precision of the method were evaluated by analysis of a blood reference material (Seronorm™ trace elements whole blood L3). Quantitative results were obtained for the determination of P, S, Cl, K and Fe, and limits of detection for these elements were adequate, taking into account their typical concentrations in real blood samples. Determination of Na, Ca, Cu and Zn was hampered by the occurrence of high sample support (Na, Ca) and instrumental blanks (Cu, Zn). Therefore, the quantitative determination of these elements at the levels expected in blood samples was not feasible. The methodology developed was applied to the analysis of several blood samples and the results obtained were compared with those reported by standard techniques. Overall, the performance of the method developed is promising and it could be used to determine the aforementioned elements in blood samples in a simple, fast and economic way. Furthermore, its non-destructive nature enables further analyses by means of complementary techniques to be carried out.

  13. Comparison of Hepatitis C Virus RNA and antibody detection in dried blood spots and plasma specimens.

    Science.gov (United States)

    Dokubo, E Kainne; Evans, Jennifer; Winkelman, Valerie; Cyrus, Sherri; Tobler, Leslie H; Asher, Alice; Briceno, Alya; Page, Kimberly

    2014-04-01

    Current diagnostic tests for Hepatitis C Virus (HCV) involve phlebotomy and serologic testing for HCV antibodies (anti-HCV) and RNA, which are not always feasible. Dried blood spots (DBS) present a minimally invasive sampling method and are suitable for sample collection, storage and testing. To assess the utility of DBS in HCV detection, we evaluated the sensitivity and specificity of DBS for anti-HCV and HCV RNA detection compared to plasma specimens. This cross-sectional validation study was conducted in the context of an existing prospective study of HCV in young injection drug users. Blood samples were collected by venipuncture into serum separator tubes (SST) and via finger stick onto Whatman 903(®) protein-saver cards. Plasma samples and eluates from the DBS were tested for anti-HCV using either a third generation enzyme-linked or chemiluminescent immunoassay (IA), and HCV RNA using discriminatory HCV transcription-mediated amplification assay (dHCV TMA). DBS results were compared to their corresponding plasma sample results. 148 participants were tested for anti-HCV and 132 participants were tested for HCV RNA. For anti-HCV, the sensitivity of DBS was 70%, specificity was 100%, positive predictive value (PPV) was 100%, negative predictive value (NPV) was 76% and Kappa was 0.69. For HCV RNA, the sensitivity of DBS was 90%, specificity was 100%, PPV was 100%, NPV was 94% and Kappa was 0.92. DBS are sensitive and very specific in detecting anti-HCV and HCV RNA, demonstrate good correlation with plasma results, and have potential to facilitate diagnosis of HCV infection. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Blood sampling from adrenal gland vein

    International Nuclear Information System (INIS)

    Sun Yong; Ni Caifang

    2009-01-01

    Adrenal gland vein sampling is an interventional method to get the blood samples from the adrenal gland vein. The blood is obtained via a catheter which is selectively inserted in the adrenal gland vein. This technique is mainly used to be diagnostic for primary hyperaldosteronism. A full knowledge of the anatomy and variations of the adrenal gland vein, serious preoperative preparation and skilled catheterization manipulation are necessary for obtaining sufficient blood sample and for reducing the occurrence of complications. Providing the physicians with definite diagnostic evidence and being technically feasible, adrenal gland vein sampling should become one of the routine examinations for clarifying the cause of primary hyperaldosteronism. (authors)

  15. Determination of glucose levels using dried filter paper blood spots: new perspective in home monitoring

    OpenAIRE

    Ward, Laura Sterian; Novis, Renata Britto; Nascimento, Veridiana Toledo; Nóbrega, Miriam Siesler; Saad, Mário José Abdalla

    1996-01-01

    We present a method for the determination of blood glucose using dried filter paper blood spots.To validate this method, we compared our results using filter paper and simultaneously collected venous blood. We demonstrated that there is a linear relationship between the filter paper glucose levels and those determined in whole blood (r=0.98). There was no significant difference between the results of the two methods (p>0.05).This method is a cheap alternative which may improve the control of ...

  16. Determination of glucose levels using dried filter paper blood spots: new perspective in home monitoring

    Directory of Open Access Journals (Sweden)

    Laura Sterian Ward

    Full Text Available We present a method for the determination of blood glucose using dried filter paper blood spots.To validate this method, we compared our results using filter paper and simultaneously collected venous blood. We demonstrated that there is a linear relationship between the filter paper glucose levels and those determined in whole blood (r=0.98. There was no significant difference between the results of the two methods (p>0.05.This method is a cheap alternative which may improve the control of diabetes mellitus, and may also be very useful in the diagnosis of postprandial hypoglycemia and other special situations.

  17. Preanalytical Blood Sampling Errors in Clinical Settings

    International Nuclear Information System (INIS)

    Zehra, N.; Malik, A. H.; Arshad, Q.; Sarwar, S.; Aslam, S.

    2016-01-01

    Background: 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. Methods: 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. Results: When asked in the questionnaire, around 61.6 percent of the paramedical staff stated that they cleaned the vein by moving the alcohol swab from inward to outwards while 20.8 percent of the staff reported that they felt the vein after disinfection. On contrary to WHO guidelines, 89.6 percent 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 percent thought that they had ample knowledge regarding the blood sampling process but they did not practice it properly. Conclusion: 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. (author)

  18. Fetal scalp blood sampling during labor

    DEFF Research Database (Denmark)

    Chandraharan, Edwin; Wiberg, Nana

    2014-01-01

    Fetal cardiotocography is characterized by low specificity; therefore, in an attempt to ensure fetal well-being, fetal scalp blood sampling has been recommended by most obstetric societies in the case of a non-reassuring cardiotocography. The scientific agreement on the evidence for using fetal...... scalp blood sampling to decrease the rate of operative delivery for fetal distress is ambiguous. Based on the same studies, a Cochrane review states that fetal scalp blood sampling increases the rate of instrumental delivery while decreasing neonatal acidosis, whereas the National Institute of Health...... and Clinical Excellence guideline considers that fetal scalp blood sampling decreases instrumental delivery without differences in other outcome variables. The fetal scalp is supplied by vessels outside the skull below the level of the cranial vault, which is likely to be compressed during contractions...

  19. Application of nuclear magnetic resonance spectroscopy combined with principal component analysis in detecting inborn errors of metabolism using blood spots: a metabonomic approach

    International Nuclear Information System (INIS)

    Constantinou, M.A.; Papakonstantinou, E.; Benaki, D.; Spraul, M.; Shulpis, K.; Koupparis, M.A.; Mikros, E.

    2004-01-01

    NMR spectra of extracted blood spots were used to investigate the possibility for the development of a new method for mass screening concerning the diagnosis of inborn errors of metabolism (IEM). Blood spots were collected on filter papers from normal, phenylketonuric (PKU) and maple syrup urine disease (MSUD) subjects and their Carr-Purcell-Meiboom-Gill (CPMG) 1 H NMR spectra were acquired. The spectra were reduced to a number of spectral descriptors and principal component analysis (PCA) was performed. The scores plot showed that PKU and MSUD samples were well discriminated from the main cluster of points

  20. Use of dried spots of whole blood, plasma, and mother's milk collected on filter paper for measurement of human immunodeficiency virus type 1 burden

    NARCIS (Netherlands)

    Ayele, Workenesh; Schuurman, Rob; Messele, Tsehaynesh; Dorigo-Zetsma, Wendelien; Mengistu, Yohannes; Goudsmit, Jaap; Paxton, William A.; de Baar, Michel P.; Pollakis, Georgios

    2007-01-01

    We studied the use of dried spots of bodily fluids (plasma, whole blood, and mother's milk) on filter paper as a means of sample collection and storage for human immunodeficiency virus type 1 (HIV-1) viral load testing under stringent field conditions. Plasma placed directly in lysis buffer, which

  1. Cytomegalovirus antibodies in dried blood spots: a minimally invasive method for assessing stress, immune function, and aging

    Directory of Open Access Journals (Sweden)

    Chyu Laura

    2011-01-01

    Full Text Available Abstract Background Cytomegalovirus (CMV is a prevalent herpesvirus with links to both stress and aging. This paper describes and validates a minimally invasive method for assessing antibodies against CMV in finger stick whole blood spot samples for use as an indirect marker of an aspect of cell-mediated immunity. Results Analysis of CMV in dried blood spot samples (DBS was based on modifications of a commercially available protocol for quantifying CMV antibodies in serum or plasma. The method was evaluated through analysis of precision, reliability, linearity, and correlation between matched serum and DBS samples collected from 75 volunteers. Correlation between DBS and plasma values was linear and high (Pearson correlation R = .96, and precision, reliability, and linearity of the DBS assay were within acceptable ranges. Conclusions The validity of a DBS assay for CMV antibodies will enable its inclusion in population-based surveys and other studies collecting DBS samples in non-clinical settings, increasing scientific understanding of the interaction of social and biological stress and immune function.

  2. Blood Sample Transportation by Pneumatic Transportation Systems

    DEFF Research Database (Denmark)

    Nybo, Mads; Lund, Merete E; Titlestad, Kjell

    2018-01-01

    BACKGROUND: Pneumatic transportation systems (PTSs) are increasingly used for transportation of blood samples to the core laboratory. Many studies have investigated the impact of these systems on different types of analyses, but to elucidate whether PTSs in general are safe for transportation...... of blood samples, existing literature on the subject was systematically assessed. METHODS: A systematic literature review was conducted following the preferred reporting items for systematic reviews and metaanalyses (PRISMA) Statement guidelines to gather studies investigating the impact of PTS on analyses...... in blood samples. Studies were extracted from PubMed and Embase. The search period ended November 2016. RESULTS: A total of 39 studies were retrieved. Of these, only 12 studies were conducted on inpatients, mainly intensive care unit patients. Blood gases, hematology, and clinical chemistry were well...

  3. The development and validation of dried blood spots for external quality assurance of syphilis serology

    Directory of Open Access Journals (Sweden)

    Smit Pieter W

    2013-02-01

    Full Text Available Abstract Background Syphilis causes up to 1,500,000 congenital syphilis cases annually. These could be prevented if all pregnant women were screened, and those with syphilis treated with a single dose of penicillin before 28 weeks gestation. In recent years, rapid point-of-care tests have allowed greater access to syphilis screening, especially in rural or remote areas, but the lack of quality assurance of rapid testing has been a concern. We determined the feasibility of using dried blood spots (DBS as specimens for quality assurance of syphilis serological assays. Methods We developed DBS extraction protocols for use with Treponema pallidum particle agglutination assay (TPPA, Treponema pallidum haemagglutination assay (TPHA and an enzyme immunoassay (EIA and compared the results with those using matching plasma samples from the same patient. Results Since DBS samples showed poor performance with TPHA and EIA (TPHA sensitivity was 50.5% (95% confidence interval: 39.9–61.2% and EIA specificity was 50.4% (95% CI: 43.7–57.1%, only the DBS TPPA was used in the final evaluation. DBS TPPA showed an sensitivity of 95.5% (95% CI: 91.3–98.0% and a specificity of 99.0% (95% CI: 98.1–99.5% compared to TPPA using plasma samples as a reference. Conclusion DBS samples can be recommended for use with TPPA, and may be of value for external quality assurance of point-of-care syphilis testing.

  4. Prenatal Education of Parents About Newborn Screening and Residual Dried Blood Spots: A Randomized Clinical Trial.

    Science.gov (United States)

    Botkin, Jeffrey R; Rothwell, Erin; Anderson, Rebecca A; Rose, Nancy C; Dolan, Siobhan M; Kuppermann, Miriam; Stark, Louisa A; Goldenberg, Aaron; Wong, Bob

    2016-06-01

    Research clearly indicates that current approaches to newborn blood spot screening (NBS) education are ineffective. Incorporating NBS education into prenatal care is broadly supported by lay and professional opinion. To determine the efficacy and effect of prenatal education about newborn screening and use of residual dried blood spots (DBS) in research on parental knowledge, attitudes, and behaviors. A randomized clinical trial of prenatal educational interventions, with outcomes measured by survey at 2 to 4 weeks postpartum. Participants were recruited from obstetric clinics in Salt Lake City, Utah; San Francisco, California; and the Bronx, New York. Eligible women were English- or Spanish-speaking adults and did not have a high-risk pregnancy. A total of 901 women were enrolled. Participants who completed the follow-up survey included 212 women in the usual care group (70% retention), 231 in the NBS group (77% retention), and 221 women in the NBS + DBS group (75% retention). Those who completed the survey were similar across the 3 groups with respect to age, ethnicity, race, education, marital status, income, obstetric history, and language. Participants were randomized into 1 of 3 groups: usual care (n = 305), those viewing an NBS movie and brochure (n = 300), and those viewing both the NBS and DBS movies and brochures (n = 296). Two to four weeks postpartum, women completed a 91-item survey by telephone, addressing knowledge, attitudes, and behavior with respect to opting out of NBS or DBS for their child. A total of 901 women (mean age, 31 years) were randomized and 664 completed the follow-up survey. The total correct responses on the knowledge instrument in regard to NBS were 69% in the usual care group, 79% in the NBS group, and 75% in the NBS + DBS group, a significant between-group difference (P women who were "very supportive" was highest in the NBS group (94%), followed by the NBS + DBS group (86%) and was lowest in the usual care

  5. Comparison of cross-sectional HIV incidence assay results from dried blood spots and plasma.

    Directory of Open Access Journals (Sweden)

    Katherine E Schlusser

    Full Text Available Assays have been developed for cross-sectional HIV incidence estimation using plasma samples. Large scale surveillance programs are planned using dried blood spot (DBS specimens for incidence assessment. However, limited information exists on the performance of HIV cross-sectional incidence assays using DBS.The assays evaluated were: Maxim HIV-1 Limiting Antigen Avidity EIA (LAg-Avidity, Sedia HIV-1 BED-Capture EIA (BED-CEIA, and CDC modified BioRad HIV-1/2 Plus O Avidity-based Assay (CDC-BioRad Avidity using pre-determined cutoff values. 100 matched HIV-1 positive plasma and DBS samples, with known duration of infection, from the Consortium for the Evaluation and Performance of HIV Incidence Assays repository were tested. All assays were run in duplicate. To examine the degree of variability within and between results for each sample type, both categorical and continuous results were analyzed. Associations were assessed with Bland Altman, R2 values and Cohen's kappa coefficient (ĸ.Intra-assay variability using the same sample type was similar for all assays (R2 0.96 to 1.00. The R2 values comparing DBS and plasma results for LAg-Avidity, BED-CEIA, and CDC-BioRad Avidity were 0.96, 0.94, and 0.84, respectively. The concordance and ĸ values between DBS and plasma for all three assays were >87% and >0.64, respectively. The Bland-Altman analysis showed significant differences between plasma and DBS samples. For all three assays, a higher number of samples were classified as recent infections using DBS samples.DBS and plasma sample results were highly correlated. However, when compared to plasma, each assay performed somewhat differently in DBS at the lower and higher ends of the dynamic range. DBS samples were more likely to be classified as recently infected by all three assays, which may lead to overestimation of incidence in surveys using performance criteria derived for plasma samples.

  6. [Comparison of arterial blood sample kits].

    Science.gov (United States)

    Calaf, N; Giner, J; Codina, E; Feixas, T; González, M; Casan, P

    2004-08-01

    Most inaccuracies in the analysis of gases and electrolytes in arterial blood samples are due to preanalytic factors, among which is the type of equipment used for blood collection. Our objective was to compare arterial blood gas sample kits used under clinical conditions and to evaluate the impact of delay in estimation on variability in results. In 2 types of study we compared 5 kits (Radiometer's Pico 70, Becton Dickinson's Preset, SIMS Portex's Pro-Vent, SIMS-Concord's Pulsator, and Marquest's Quick ABG). In the first study kitsyringe assignment was randomized for collecting arterial blood samples from 160 consecutive patients to evaluate practical aspects of using them and the presence of bubbles in the samples taken. The second study evaluated the effects of delays of 30 and 60 minutes in estimation and of the type of heparin used in 54 blood samples. The kits which produced the fewest bubbles, gave samples with the greatest stability, and had the least impact on ion concentration were Radiometer's Pico 70 and SIMS-Portex's Pro-vent.

  7. Isolation of human genomic DNA for genetic analysis from premature neonates: a comparison between newborn dried blood spots, whole blood and umbilical cord tissue

    Science.gov (United States)

    2013-01-01

    Background Genotyping requires biological sample collection that must be reliable, convenient and acceptable for patients and clinicians. Finding the most optimal procedure of sample collection for premature neonates who have a very limited blood volume is a particular challenge. The aim of the current study was to evaluate the use of umbilical cord (UC) tissue and newborn dried blood spot (DBS)-extracted genomic DNA (gDNA) as an alternative to venous blood-derived gDNA from premature neonates for molecular genetic analysis. All samples were obtained from premature newborn infants between 24-32 weeks of gestation. Paired blood and UC samples were collected from 31 study participants. gDNA was extracted from ethylenediaminetetraacetic acid (EDTA) anticoagulant-treated blood samples (~500 μl) and newborn DBSs (n = 723) using QIAamp DNA Micro kit (Qiagen Ltd., Crawley, UK); and from UC using Qiagen DNAeasy Blood and Tissue kit (Qiagen Ltd., Crawley, UK). gDNA was quantified and purity confirmed by measuring the A260:A280 ratio. PCR amplification and pyrosequencing was carried out to determine suitability of the gDNA for molecular genetic analysis. Minor allele frequency of two unrelated single nucleotide polymorphisms (SNPs) was calculated using the entire cohort. Results Both whole blood samples and UC tissue provided good quality and yield of gDNA, which was considerably less from newborn DBS. The gDNA purity was also reduced after 3 years of storage of the newborn DBS. PCR amplification of three unrelated genes resulted in clear products in all whole blood and UC samples and 86%-100% of newborn DBS. Genotyping using pyrosequencing showed 100% concordance in the paired UC and whole blood samples. Minor allele frequencies of the two SNPs indicated that no maternal gDNA contamination occurred in the genotyping of the UC samples. Conclusions gDNAs from all three sources are suitable for standard PCR and pyrosequencing assays. Given that UC provide good quality

  8. Estimation of HIV-1 DNA Level Interfering with Reliability of HIV-1 RNA Quantification Performed on Dried Blood Spots Collected from Successfully Treated Patients

    OpenAIRE

    Zida, Sylvie; Tuaillon, Edouard; Barro, Makoura; Kwimatouo Lekpa Franchard, Arnaud; Kagoné, Thérèse; Nacro, Boubacar; Ouedraogo, Abdoul Salam; Bolloré, Karine; Sanosyan, Armen; Plantier, Jean-Christophe; Meda, Nicolas; Sangaré, Lassana; Rouzioux, Christine; Rouet, François; Kania, Dramane

    2016-01-01

    The impact of HIV-1 DNA coamplification during HIV-1 RNA quantification on dried blood spots (DBS) was explored. False-positive HIV RNA detection (22/62, 35%) was associated with high HIV-1 DNA levels. Specificity of HIV-1 RNA assays on DBS should be evaluated following manufacturer protocols on samples with HIV-1 DNA levels of ≥1,000 copies/106 peripheral blood mononuclear cells.

  9. Evaluation of market samples of ′Yashada bhasma′ using ′Namburi Phased Spot Test′

    Directory of Open Access Journals (Sweden)

    Santhosh Bhojashettar

    2011-01-01

    Full Text Available Yashada bhasma (Calx of Yashada i.e. Zinc which has its main indication in Prameha (Diabetes and Netra vikaras (Eye disorders was prepared according to the prescription in the Ayurvedic classics and subjected to various bhasma parikshas, including the Namburi Phased Spot Test (NPST, one of the qualitative tests described for various Ayurvedic preparations. NPST helps differentiate between, and thus identify, various bhasmas. It depends upon the pattern of the spot, which develops after a specific chemical reaction. Three market samples of Yashada bhasma, which were said to be Parada marita (incinerated using Mercury, were also subjected to the above tests and results compared. The various bhasmas exhibited marked differences in colour, and though NPST yielded desired results for all the samples, there were differences in their spot patterns and colour. The bhasma prepared in our department produced the most accurate results.

  10. Determination of gemifloxacin on dried blood spots by hydrophilic interaction liquid chromatography with fluorescence detector: application to pharmacokinetics in rats.

    Science.gov (United States)

    Nageswara Rao, R; Naidu, Ch Gangu; Guru Prasad, K; Padiya, Raju; Agwane, Sachin B

    2012-12-01

    A highly selective, sensitive and rapid hydrophilic liquid interaction chromatographic method was developed and validated for determination of gemifloxacin on dried blood spots. The chromatographic separation was achieved on a reversed-phase zwitterionic hydrophilic interaction liquid chromatographic ZIC®HILIC-C₁₈ (4.6 × 100 mm; 5 µm) column using acetonitrile-10 mM ammonium acetate (pH 3.5; 80:20, v/v) as a mobile phase in an isocratic elution mode at a flow rate 0.6 mL/min at 27 °C. An on-line fluorescence detector set at excitation and emission wavelengths of 269 and 393 nm, respectively was used for monitoring column eluents. Ciprofloxacin was used as an internal standard. The method was validated for accuracy, precision, linearity and selectivity by design of experiments following ICH guidelines. The assay exhibited a linear range of 25-5000 ng/mL for gemifloxacin on dried blood spots. The lower limit of detection was found to be 10 ng/mL. The intra- and inter-assay coefficients of variation did not exceed 7.4% deviation of the nominal concentration. The recovery of GFX from dried blood spots was >95.0% and its stability was excellent with no evidence of degradation during sample processing for at least 3 months storage in a freezer at -20 °C. Copyright © 2012 John Wiley & Sons, Ltd.

  11. Reduction of Radiation Exposure Using Dynamic Trace Digital Angiography and Spot Fluoroscopy During Adrenal Venous Sampling.

    Science.gov (United States)

    Morita, Satoru; Endo, Kenji; Suzaki, Shingo; Ishizaki, Umiko; Yamazaki, Hiroshi; Nishina, Yu; Sakai, Shuji

    2017-05-01

    To compare radiation exposure of adrenal venous sampling (AVS) using dynamic trace digital angiography (DTDA) and spot fluoroscopy with that using conventional methods. AVS was performed in 11 patients using DTDA and spot fluoroscopy (Group A) and 11 patients using conventional digital subtraction angiography (DSA) with collimation (Group B). Radiation exposure and image quality of adrenal venography using a five-point scale were compared between the groups. The acquisition dose-area product (DAP) using DTDA and fluoro-DAP using spot fluoroscopy in Group A were lower than those using conventional DSA (5.3 ± 3.7 vs. 29.1 ± 20.1 Gy cm 2 , p fluoroscopy without sacrificing image quality.

  12. Improving blood sample logistics using simulation

    DEFF Research Database (Denmark)

    Jørgensen, Pelle Morten Thomas; Jacobsen, Peter

    2012-01-01

    Using simulation as an approach to display and improve internal logistics and handling at hospitals has great potential. This research will show how a simulation model can be used to evaluate changes made to two different cases of transportation of blood samples at a hospital, by evaluating...

  13. Automated system for fractionation of blood samples

    Energy Technology Data Exchange (ETDEWEB)

    Lee, N. E.; Genung, R. K.; Johnson, W. F.; Mrochek, J. E.; Scott, C. D.

    1978-01-01

    A prototype system for preparing multiple fractions of blood components (plasma, washed red cells, and hemolysates) using automated techniques has been developed. The procedure is based on centrifugal separation and differential pressure-induced transfer in a rotor that has been designed to process numerous samples simultaneously. Red cells are sedimented against the outer walls of the sample chamber, and plasma is syphoned, by imposition of eithr a slight positive or negative pressure, into individual reservoirs in a collection ring. Washing of cells is performed in situ; samples of washed cells, either packed or in saline solution, can be recovered. Cellular hemolysates are prepared and automatically transferred to individual, commercially available collection vials ready for storage in liquid nitrogen or immediate analysis. The system has potential application in any biomedical area which requires high sample throughput and in which one or more of the blood fractions will be used. A separate unit has been designed and developed for the semiautomated cleaning of the blood processing vessel.

  14. Measurement and application of purine derivatives: Creatinine ratio in spot urine samples of ruminants

    International Nuclear Information System (INIS)

    Chen, X.B.; Jayasuriya, M.C.N.; Makkar, H.P.S.

    2004-01-01

    The daily excretion of purine derivatives in urine has been used to estimate the supply of microbial protein to ruminant animals. The method provides a simple and non-invasive tool to indicate the nutritional status of farm animals. However due to the need for complete collection of urine the potential application at farm level is restricted. Research conducted under the FAO/IAEA Co-ordinated Research Project has indicated that it is possible to use the purine derivatives:creatinine ratio measured in several spot urine samples collected within a day, as an index of microbial protein supply in a banding system for farm application. Some theoretical and experimental aspects in the measurement of purine derivatives:creatinine ratio in spot urine samples and the possible application of the banding system at the farm level are discussed. (author)

  15. Automated blood sampling systems for positron emission tomography

    International Nuclear Information System (INIS)

    Eriksson, L.; Holte, S.; Bohm, C.; Kesselberg, M.; Hovander, B.

    1988-01-01

    An automated blood sampling system has been constructed and evaluated. Two different detector units in the blood sampling system are compared. Results from studies of blood-brain barrier transfer of a C-11 labelled receptor antagonist will be discussed

  16. Streamlined determination of lysophosphatidylcholines in dried blood spots for newborn screening of X-linked adrenoleukodystrophy.

    Science.gov (United States)

    Turgeon, Coleman T; Moser, Ann B; Mørkrid, Lars; Magera, Mark J; Gavrilov, Dimitar K; Oglesbee, Devin; Raymond, Kimiyo; Rinaldo, Piero; Matern, Dietrich; Tortorelli, Silvia

    2015-01-01

    Pre-symptomatic hematopoietic stem cell transplantation is essential to achieve best possible outcomes for patients with the childhood cerebral form of X-linked adrenoleukodystrophy (X-ALD). We describe a high-throughput method for measurement of C20-C26 lysophosphatidylcholines (LPCs) and biochemical diagnosis of X-ALD using the same dried blood spots (DBS) routinely used for newborn screening. LPCs are extracted from 3-mm DBS punch with methanol containing an isotopically labeled LPC as internal standard. This extract is transferred to a 96-well plate, evaporated and then reconstituted in mobile phase for flow injection analysis tandem mass spectrometry (FIA-MS/MS) in selected reaction monitoring mode for measurement of four different LPCs (C20, C22, C24, C26) and the internal standard (d4-C26-LPC). Analysis time is 1.5min per sample. The mean CVs from the intra- and inter-assay experiments for LPCs were 6.3-15.1% for C20-LPC, 4.4-18.6% for C22-LPC and 4.5-14.3% for C24-LPC. Limits of detection were determined for C20-LPC (LOD=0.03μg/mL), C22-LPC (0.03μg/mL), C24-LPC (0.03μg/mL) and C26-LPC (0.01μg/mL). Reference ranges were established from DBS of 130 newborns and 20 adults. Samples of patients with X-ALD (n=16), peroxisomal biogenesis disorders (n=8), and X-ALD carriers (n=12) were analyzed blindly and all were correctly identified. Analysis of LPC species by FIA-MS/MS is a fast, simple and reliable method to screen for X-ALD and other peroxisomal disorders in DBS. To maximize specificity, abnormal results can be verified by a 2nd tier assay using LC-MS/MS. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. The influence of the dried blood spot drying time on the recoveries of six immunosuppressants

    Directory of Open Access Journals (Sweden)

    Remco A. Koster

    2015-10-01

    Full Text Available Investigation of the drying time of dried blood spots (DBS is currently not included in DBS validations. The influence of the DBS drying time on the recovery of tacrolimus, ascomycin, sirolimus, everolimus, cyclosporin A and temsirolimus was evaluated by measuring DBS with a fixed blood volume at a hematocrit range between 0.1 and 0.6 L/L at 3, 24 and 48 hours of drying time. Results showed that the recovery of sirolimus, everolimus, temsirolimus and cyclosporin A was influenced by the DBS drying time, while the recovery of tacrolimus and ascomycin was not. A drying time of at least 24 hours is advised in order to stabilize hematocrit and concentration related recovery effects of sirolimus, everolimus, temsirolimus and cyclosporin A.

  18. 21 CFR 868.1100 - Arterial blood sampling kit.

    Science.gov (United States)

    2010-04-01

    ...) 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 samples... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Arterial blood sampling kit. 868.1100 Section 868...

  19. Comparison of calcium, magnesium, sodium, potassium, zinc, and creatinine concentration in 24-h and spot urine samples in women.

    Science.gov (United States)

    Ilich, Jasminka Z; Blanusa, Maja; Orlić, Zeljka Crncević; Orct, Tatjana; Kostial, Krista

    2009-01-01

    The 24-h urine sample is considered as the most reliable material for testing many but not necessarily all constituents in urine. However, its collection is tedious for both patients and research participants. The aim of this study was to compare concentrations of essential elements calcium (Ca), magnesium (Mg), sodium (Na), potassium (K), and zinc (Zn) in 24-h and spot urine samples. Urine samples were collected from 143 generally healthy women, aged 30-79 years. Fasting spot urine was collected immediately after the end of the 24-h collection, therefore being of the same content as the first morning urine which ended the 24-h collection. Elements were analyzed by flame atomic absorption/emission spectrometry and expressed as mg/g and/or mmol/mol of creatinine (Cr). Spearman rank order correlations between 24-h and spot urine were carried out for each element. Ratios of elements in 24-h to spot urine samples were calculated to estimate the element-proportion of spot in the 24-h sample. All coefficients of correlation between 24-h and spot urine of measured elements and Cr were significant (purine samples for each element (except K) were similar and close to 2, indicating uniform proportion of elements from spot urine sample in the 24-h sample. In addition, a high correlation between various pairs of elements was obtained in both 24-h and spot urine; the highest being between Na/Ca (0.435) and (0.578), respectively. This is in accordance with theoretical presumptions and previous findings regarding those relationships. Although replacing burdensome 24-h urine collection with spot urine sampling might not provide the solution in all cases, our results show that for the elements analyzed, spot urine could be a reliable alternative.

  20. DEVELOPMENT OF REAL-TIME MULTIPLEX PCR FOR THE QUANTITATIVE DETERMINATION OF TREC'S AND KREC'S IN WHOLE BLOOD AND IN DRIED BLOOD SPOTS

    Directory of Open Access Journals (Sweden)

    M. A. Gordukova

    2015-01-01

    Full Text Available Primary immunodeficiencies (PID such as severe combined immunodeficiency (SCID and X-linked agammaglobulinemia are characterized by the lack of functional Tand B-cells, respectively. Without early diagnosis and prompt treatment children with PID suffer from severe infectious diseases, leading to their death or disability. Our purpose was developing of simple, inexpensive, high throughput technique based on the quantitative determination of TREC and KREC molecules by real-time PCR, and its validation in a group of children with a verified diagnosis of SCID and X-linked agammaglobulinemia.In this study, we developed and validated multiplex real-time PCR for the TREC’s and KREC’s quantitative analysis. We have shown that linear range of Ct changes depending on the concentrations of targets with a correlation coefficient R2 not worse than 0.98 was observed at concentrations from 109 to 5 × 104 copies per ml. The lowest amount of targets reliably detected in a reaction volume was 10 TREC’s copies, 5 KREC ‘s copies and 5 copies of internal control (IL17RA. We determined the age-depended reference values of TRECs and KRECs in whole blood in 29 boys and 27 girls with normal immunological parameters. The normal cut-offs for TRECs and KRECs were defined in dry blood spots depending on the method of extraction.The proposed method showed 100% diagnostic sensitivity and specificity in the studied group. The method can be proposed as a screening tool for the diagnosis of SCID and X-linked agammaglobulinemia both in whole blood and in the dry blood spots. The further investigation is required with larger number of samples

  1. Technical Stability and Biological Variability in MicroRNAs from Dried Blood Spots: A Lung Cancer Therapy-Monitoring Showcase.

    Science.gov (United States)

    Kahraman, Mustafa; Laufer, Thomas; Backes, Christina; Schrörs, Hannah; Fehlmann, Tobias; Ludwig, Nicole; Kohlhaas, Jochen; Meese, Eckart; Wehler, Thomas; Bals, Robert; Keller, Andreas

    2017-09-01

    Different work flows have been proposed to use miRNAs as blood-borne biomarkers. In particular, the method used for collecting blood from patients can considerably influence the diagnostic results. We explored whether dried blood spots (DBSs) facilitate stable miRNA measurements and compared its technical stability with biological variability. First, we tested the stability of DBS samples by generating from 1 person 18 whole-genome-wide miRNA profiles of DBS samples that were exposed to different temperature and humidity conditions. Second, we investigated technical reproducibility by performing 7 replicates of DBS again from 1 person. Third, we investigated DBS samples from 53 patients with lung cancer undergoing different therapies. Across these 3 stages, 108 genome-wide miRNA profiles from DBS were generated and evaluated biostatistically. In the stability analysis, we observed that temperature and humidity had an overall limited influence on the miRNomes (average correlation between the different conditions of 0.993). Usage of a silica gel slightly diminished DBS' technical reproducibility. The 7 technical replicates had an average correlation of 0.996. The correlation with whole-blood PAXGene miRNomes of the same individual was remarkable (correlation of 0.88). Finally, evaluation of the samples from the 53 patients with lung cancer exposed to different therapies showed that the biological variations exceeded the technical variability significantly ( P technical variations significantly. DBS-based miRNA profiles will potentially further the translational character of miRNA biomarker studies. © 2017 American Association for Clinical Chemistry.

  2. Population variability of phthalate metabolites and bisphenol A concentrations in spot urine samples versus 24- or 48-h collections.

    Science.gov (United States)

    Christensen, Krista L Yorita; Lorber, Matthew; Koch, Holger M; Kolossa-Gehring, Marike; Morgan, Marsha K

    2012-11-01

    Human exposure to phthalates and bisphenol A (BPA) can be assessed through urinary biomonitoring, but methods to infer daily intakes assume that spot sample concentrations are comparable to daily average concentrations. We evaluate this assumption using human biomonitoring data from Germany and the United States (US). The German data comprised three regional studies with spot samples and one with full-day samples analyzed for phthalate metabolites. The US data included: a study on DEHP metabolites and BPA involving eight persons supplying all urine voids (from which 24-h samples were constructed) for seven consecutive days; NHANES spot sample data on DEHP metabolites and BPA; and a regional study of children with 48-h samples analyzed for BPA. In the German data, measures of central tendency differed, but spot and 24-h samples showed generally comparable variance including 95th percentiles and maxima equidistant from central tendency measures. In contrast, the US adult data from the eight-person study showed similar central tendencies for phthalate metabolites and BPA, but generally greater variability for the spot samples, including higher 95th percentiles and maxima. When comparing children's BPA concentrations in NHANES spot and 48-h samples, distributions showed similar central tendency and variability. Overall, spot urinary concentrations of DEHP metabolites and BPA have variability roughly comparable with corresponding 24-h average concentrations obtained from a comparable population, suggesting that spot samples can be used to characterize population distributions of intakes. However, the analysis also suggests that caution should be exercised when interpreting the high end of spot sample data sets.

  3. Simultaneous measurement of 25 inflammatory markers and neurotrophins in neonatal dried blood spots by immunoassay with xMAP technology

    DEFF Research Database (Denmark)

    Skogstrand, Kristin; Thorsen, Poul; Nørgaard-Pedersen, Bent

    2005-01-01

    BACKGROUND: Inflammatory reactions and other events in early life may be part of the etiology of late-onset diseases, including cerebral palsy, autism, and type 1 diabetes. Most neonatal screening programs for congenital disorders are based on analysis of dried blood spot samples (DBSS), and stored...... on flowmetric Luminex xMAP technology to measure inflammatory markers and neutrophins in DBSS. RESULTS: The high-capacity 25-plex multianalyte method measured 23 inflammatory and trophic cytokines, triggering receptor expressed on myeloid cells-1 (TREM-1), and C-reactive protein in two 3.2-mm punches from DBSS....... It also measured 26 cytokines and TREM-1 in serum. Standards Recovery in the 25-plex method were 90%-161% (mean, 105%). The low end of the working range for all 25 analytes covered concentrations found in DBSS from healthy newborns. Mean recovery of exogenous analytes added at physiologic concentrations...

  4. [Clinical application of protein-to-creatinine ratio in spot urine samples for preeclampsia].

    Science.gov (United States)

    Kuang, Lin; Liang, Feng-bing; Yang, Cui-yu

    2012-08-07

    To evaluate the clinical application of protein-to-creatinine ratio (P/Cr) in spot urine samples so as to check whether it can replace urine protein excretion in 24 h collections for the diagnosis and screening of preeclampsia. The investigators selected 100 cases of pregnant women with preeclampsia and 36 cases of normal pregnant women examined at Sir Run Run Shaw Hospital over the last two years. The correlations between P/Cr in first morning urine samples and urinary protein excretion in 24 h collections were analyzed. The cutoff values of P/Cr in first morning urine samples for screening preeclampsia of 0.3 g and 5 g in urinary protein excretion in 24 h collections were determined by a receiver operating characteristics (ROC) curve. A highly significant correlation existed between P/Cr in first morning urine samples and urinary protein excretion in 24 h collections. By the ROC curve analysis, the P/Cr of 0.34 g/g and 2.08 g/g in first morning urine samples represented the most appropriate threshold for detecting the urinary protein excretion of 0.3 g and 5 g in 24 h collections. The P/Cr in spot urine samples can replace urinary protein excretion in 24 h collections. It is a simple and reliable tool of diagnosis and follow-up for preeclampsia.

  5. Simplified sample preparation using frame spotting method for direct counting of total bacteria by fluorescence microscopy.

    Science.gov (United States)

    Maruyama, Fumito; Yamaguchi, Nobuyasu; Kenzaka, Takehiko; Tani, Katsuji; Nasu, Masao

    2004-12-01

    A new preparation method for direct counting of bacteria in liquid samples with fluorescence microscope was developed using a glass slide coated with 3-aminopropyltriethoxy silane and ring-shaped polyester seal as a retainer. The experimental steps of this method were spotting samples onto the coated slides with the seal, drying under vacuum, staining with SYBR Green II, drying and covering with immersion oil and coverslip to allow counting. This simplified method provided consistent results when compared with the conventional filtration method for fluorescence microscopy, and is rapid, inexpensive and reproducible.

  6. Detection of cytomegalovirus DNA in dried blood spots of Minnesota infants who do not pass newborn hearing screening.

    Science.gov (United States)

    Choi, K Yeon; Schimmenti, Lisa A; Jurek, Anne M; Sharon, Bazak; Daly, Kathy; Khan, Cindy; McCann, Mark; Schleiss, Mark R

    2009-12-01

    Up to 15% of infants with asymptomatic congenital cytomegalovirus (CMV) infection will experience some degree of sensorineural hearing loss. Many infants who fail newborn hearing screening (NHS) are likely to have congenital CMV infection, but may escape definitive virologic identification because diagnostic evaluation may not commence until several weeks or months of age, making differentiation between congenital and postnatal CMV infection difficult. Early diagnosis linking virologic identification of congenital CMV infection to infants failing NHS may improve diagnostic precision and enhance opportunities for therapeutic intervention. The goal of this study was to compare newborn dried blood spots from Minnesota infants who had failed NHS, and were designated for referral, with control infants who passed NHS, for the presence of CMV DNA by real-time PCR, using hybridization probes for the CMV gene UL54. Of 479 infants with a failed NHS (bilateral failure), 13 had CMV DNA present in the blood spot (2.7%). This compared with only 2/479 positive results from a control group of infants who passed the NHS (0.4%; P = 0.007, Fisher exact test). Comparisons of the glycoprotein B (gB) genotype as well as direct DNA sequencing of selected positives revealed that PCR positive samples represented unique clinical isolates. The mean viral load among the 15 positive samples was 1.6 x 10(3) genomes/microgram of total DNA. Newborn bloodspot CMV screening by real-time PCR may be a useful and rapid adjunct to functional NHS and may enable more rapid etiologic diagnosis of sensorineural hearing loss in newborns.

  7. A New Method to Quantify Ifosfamide Blood Levels Using Dried Blood Spots and UPLC-MS/MS in Paediatric Patients with Embryonic Solid Tumours.

    Directory of Open Access Journals (Sweden)

    Luz-María Torres

    Full Text Available Ifosfamide blood concentrations are necessary to monitor its therapeutic response, avoiding any adverse effect. We developed and validated an analytical method by UPLC-MS/MS to quantify ifosfamide in dried blood spots (DBS. Blood samples were collected on Whatman 903® filter paper cards. Five 3 mm disks were punched out from each dried blood spot. Acetonitrile and ethyl acetate were used for drug extraction. Chromatographic separation was carried out in an Acquity UPLC equipment with a BEH-C18 column, 2.1 x 100 mm, 1.7 μm (Waters®. The mobile phase consisted in 5 mM ammonium formate and methanol:acetonitrile (40:48:12 v/v/v at 0.2 mL/min. LC-MS/MS detection was done by ESI+ and multiple reaction mode monitoring, ionic transitions were m/z1+ 260.99 > 91.63 for ifosfamide and 261.00 > 139.90 for cyclophosphamide (internal standard. This method was linear within a 100-10000 ng/mL range and it was accurate, precise and selective. Ifosfamide samples in DBS were stable for up to 52 days at -80°C. The procedure was tested in 14 patients, ages 1 month to 17 years (9 males and 5 females, with embryonic tumours treated with ifosfamide, alone or combined, at a public tertiary referral hospital. Ifosfamide blood levels ranged from 11.1 to 39.7 μmol/L at 12 hours after the last infusion, while 24-hour levels ranged from 0.7-19.7 μmol/L. The median at 12 hours was 19.5 μmol/L (Q25 14.4-Q75 29.0 and 3.8 μmol/L (Q25 1.5-Q75 9.9 at 24 hours, p<0.001. This method is feasible to determine ifosfamide plasma levels in paediatric patients.

  8. Whole exome and whole genome sequencing with dried blood spot DNA without whole genome amplification.

    Science.gov (United States)

    Bassaganyas, Laia; Freedman, George; Vaka, Dedeepya; Wan, Eunice; Lao, Richard; Chen, Flavia; Kvale, Mark; Currier, Robert J; Puck, Jennifer M; Kwok, Pui-Yan

    2018-01-01

    Newborn screening (NBS) for rare conditions is performed in all 50 states in the USA. We have partnered with the California Department of Public Health Genetic Disease Laboratory to determine whether sufficient DNA can be extracted from archived dried blood spots (DBS) for next-generation sequencing in the hopes that next-generation sequencing can play a role in NBS. We optimized the DNA extraction and sequencing library preparation protocols for residual infant DBS archived over 20 years ago and successfully obtained acceptable whole exome and whole genome sequencing data. This sequencing study using DBS DNA without whole genome amplification prior to sequencing library preparation provides evidence that properly stored residual newborn DBS are a satisfactory source of DNA for genetic studies. © 2017 Wiley Periodicals, Inc.

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

    Science.gov (United States)

    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.

  10. High-performance liquid chromatography determination of dapsone, monoacetyldapsone, and pyrimethamine in filter paper blood spots

    DEFF Research Database (Denmark)

    Rønn, A M; Lemnge, M M; Angelo, H R

    1995-01-01

    A high-performance liquid chromatography method for the simultaneous analysis of dapsone (DDS), the major metabolite of DDS, monoacetyldapsone (MADDS), and pyrimethamine (PYR) was modified for capillary blood samples obtained by finger prick and dried on filter paper. Limit of quantitation using...... 150 microliters whole blood dried on filter paper was found to be 20 ng/ml for DDS and PYR and 15 ng/ml for MADDS (precision filter...... paper method and the original whole-blood method was almost comparable. Standardization could therefore be obtained by the more simple whole-blood method. Dried filter paper samples stored at 19-22 degrees C were stable for months and for 2 weeks stored at 35 degrees C. The concentrations...

  11. Congener Production in Blood Samples During Preparation and Storage

    DEFF Research Database (Denmark)

    Felby, Søren; Nielsen, Erik

    1995-01-01

    Retsmedicin, congener production, preparation, head space GC, acetone, isobutanol, storage, blood samples, n-propanol, methanol, methylethylketone......Retsmedicin, congener production, preparation, head space GC, acetone, isobutanol, storage, blood samples, n-propanol, methanol, methylethylketone...

  12. Reliable Quantification of the Potential for Equations Based on Spot Urine Samples to Estimate Population Salt Intake

    DEFF Research Database (Denmark)

    Huang, Liping; Crino, Michelle; Wu, Jason Hy

    2016-01-01

    to a standard format. Individual participant records will be compiled and a series of analyses will be completed to: (1) compare existing equations for estimating 24-hour salt intake from spot urine samples with 24-hour urine samples, and assess the degree of bias according to key demographic and clinical......BACKGROUND: Methods based on spot urine samples (a single sample at one time-point) have been identified as a possible alternative approach to 24-hour urine samples for determining mean population salt intake. OBJECTIVE: The aim of this study is to identify a reliable method for estimating mean...... population salt intake from spot urine samples. This will be done by comparing the performance of existing equations against one other and against estimates derived from 24-hour urine samples. The effects of factors such as ethnicity, sex, age, body mass index, antihypertensive drug use, health status...

  13. [Can we rule out a congenital cytomegalovirus infection when the result of polymerase chain reaction in dried blood spots is negative?].

    Science.gov (United States)

    Vives-Oñós, Isabel; Soler-Palacín, Pere; Codina-Grau, María Gemma; Martín-Nalda, Andrea; López-Galera, Rosa María; Marín-Soria, José Luís; Figueras-Nadal, Concepció

    2014-11-01

    The detection of cytomegalovirus (CMV) DNA by real time polymerase chain reaction (rt-PCR) in dried blood spots collected routinely for metabolic screening has been assessed for the retrospective diagnosis of congenital CMV (cCMV) infection in many studies, but not in Spain. The aim of this study is to analyze the diagnostic accuracy of this technique in our hospital. A cross-sectional retrospective observational study was conducted including all patients born between January, 2007 and September, 2012 with confirmed cCMV infection. The assessment of CMV DNA was made by using rt-PCR in dried blood spots of these patients. Fourteen patients were included: 4/14 were symptomatic and 4/14 had sequelae. The detection of CMV DNA by rt-PCR was positive in only 7 patients. A statistically significant relationship between low viral load at birth and negative rt-PCR in dried blood spots was demonstrated. Despite the low number of patients included, our data highlight an important amount of false negative results in the DNA CMV detection by rt-PCR in these samples for the retrospective diagnosis of cCMV infection, especially in cases with low viral load at birth. Copyright © 2013 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  14. Optimized DNA extraction from neonatal dried blood spots: application in methylome profiling

    Science.gov (United States)

    2014-01-01

    Background Neonatal dried blood spots (DBS) represent an inexpensive method for long-term biobanking worldwide and are considered gold mines for research for several human diseases, including those of metabolic, infectious, genetic and epigenetic origin. However, the utility of DBS is restricted by the limited amount and quality of extractable biomolecules (including DNA), especially for genome wide profiling. Degradation of DNA in DBS often occurs during storage and extraction. Moreover, amplifying small quantities of DNA often leads to a bias in subsequent data, particularly in methylome profiles. Thus it is important to develop methodologies that maximize both the yield and quality of DNA from DBS for downstream analyses. Results Using combinations of in-house-derived and modified commercial extraction kits, we developed a robust and efficient protocol, compatible with methylome studies, many of which require stringent bisulfite conversion steps. Several parameters were tested in a step-wise manner, including blood extraction, cell lysis, protein digestion, and DNA precipitation, purification and elution. DNA quality was assessed based on spectrophotometric measurements, DNA detectability by PCR, and DNA integrity by gel electrophoresis and bioanalyzer analyses. Genome scale Infinium HumanMethylation450 and locus-specific pyrosequencing data generated using the refined DBS extraction protocol were of high quality, reproducible and consistent. Conclusions This study may prove useful to meet the increased demand for research on prenatal, particularly epigenetic, origins of human diseases and for newborn screening programs, all of which are often based on DNA extracted from DBS. PMID:24980254

  15. Impact of blood sampling in very preterm infants

    DEFF Research Database (Denmark)

    Madsen, L P; Rasmussen, M K; Bjerregaard, L L

    2000-01-01

    ; the groups were then subdivided into critically ill or not. Diagnostic blood sampling and blood transfusion events were recorded. In total, 1905 blood samples (5,253 analysis) were performed, corresponding to 0.7 samples (1.9 analysis) per day per infant. The highest frequencies were found during the first....../kg. For the extremely preterm infants a significant correlation between sampled and transfused blood volume was found (mean 37.1 and 33.3 ml/kg, respectively, r = + 0.71, p = 0.0003). The most frequently requested analyses were glucose, sodium and potassium. Few blood gas analyses were requested (1.9/ infant). No blood...... losses attributable to excessive generous sampling were detected. The results show an acceptable low frequency of sampling and transfusion events for infants of GA 28-32 weeks. The study emphasizes the necessity of thorough reflection and monitoring of blood losses when ordering blood sampling...

  16. Governing biological material at the intersection of care and research: the use of dried blood spots for biobanking

    NARCIS (Netherlands)

    Douglas, C.M.W.; van El, C.G.; Faulkner, A.; Cornel, M.C.

    2012-01-01

    A series of governance issues currently surrounds the multiple uses and multiple users of dried blood spots (DBS) for research purposes. Internationally there is a discussion on storing DBS resulting from newborn screening for public health and using them as the basis for large biobanklike

  17. The Effect of Storage and Extraction Methods on Amplification of Plasmodium falciparum DNA from Dried Blood Spots

    NARCIS (Netherlands)

    Schwartz, A.; Baidjoe, A.Y.; Rosenthal, P.J.; Dorsey, G.; Bousema, T.; Greenhouse, B.

    2015-01-01

    Extraction and amplification of DNA from dried blood spots (DBS) collected in field studies is commonly used for detection of Plasmodium falciparum. However, there have been few systematic efforts to determine the effects of storage and extraction methods on the sensitivity of DNA amplification. We

  18. Dried blood spots: liquid chromatography-mass spectrometry analysis of Δ9-tetrahydrocannabinol and its main metabolites.

    Science.gov (United States)

    Mercolini, Laura; Mandrioli, Roberto; Sorella, Vittorio; Somaini, Lorenzo; Giocondi, Daniele; Serpelloni, Giovanni; Raggi, Maria Augusta

    2013-01-04

    A sensitive and selective HPLC-MS/MS method has been developed for the first time for the analysis of Δ(9)-tetrahydrocannabinol (the most important active cannabinoid) and its hydroxylated and carboxylated metabolites in human Dried Blood Spots (DBSs). The simultaneous determination of Δ(9)-tetrahydrocannabinol and its two main metabolites allows assessing the time elapsed after the drug intake and distinguishing between acute or former consumption. This is an important information in specific contexts such as "on street" controls by police forces. DBSs have been chosen as the optimal biological matrix for this kind of testing, since they provide information on the actual state of intoxication, without storage and transportation problems usually associated with classical blood testing. The analysis is carried out on a C8 reversed phase column with a mobile phase composed of 0.1% formic acid in a water/methanol mixture and an electrospray ionisation (ESI) source, coupled to a triple quadrupole mass spectrometer. The method was validated according to international guidelines, with satisfactory results in terms of extraction yields, precision, stability and accuracy. Application to real DBS samples from Cannabis abusers gave reliable results, thus confirming the methodology suitability for roadside testing. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. Reduction of Radiation Exposure Using Dynamic Trace Digital Angiography and Spot Fluoroscopy During Adrenal Venous Sampling

    Energy Technology Data Exchange (ETDEWEB)

    Morita, Satoru, E-mail: i@imodey.com; Endo, Kenji; Suzaki, Shingo; Ishizaki, Umiko; Yamazaki, Hiroshi; Nishina, Yu; Sakai, Shuji [Tokyo Women’s Medical University Hospital, Department of Diagnostic Imaging and Nuclear Medicine (Radiology) (Japan)

    2017-05-15

    PurposeTo compare radiation exposure of adrenal venous sampling (AVS) using dynamic trace digital angiography (DTDA) and spot fluoroscopy with that using conventional methods.Materials and MethodsAVS was performed in 11 patients using DTDA and spot fluoroscopy (Group A) and 11 patients using conventional digital subtraction angiography (DSA) with collimation (Group B). Radiation exposure and image quality of adrenal venography using a five-point scale were compared between the groups.ResultsThe acquisition dose–area product (DAP) using DTDA and fluoro-DAP using spot fluoroscopy in Group A were lower than those using conventional DSA (5.3 ± 3.7 vs. 29.1 ± 20.1 Gy cm{sup 2}, p < 0.001) and collimation (33.3 ± 22.9 vs. 59.1 ± 35.7 Gy cm{sup 2}, p = 0.088) in Group B. The total DAP in Group A was significantly lower than that in Group B (38.6 ± 25.9 vs. 88.2 ± 53.6 Gy cm{sup 2}, p = 0.006). The peak skin dose for patients and operator radiation exposure in Group A were significantly lower than those in Group B (403 ± 340 vs. 771 ± 416 mGy, p = 0.030, and 17.1 ± 14.8 vs. 36.6 ± 21.7 μSv, p = 0.013). The image quality of DTDA (4.4 ± 0.6) was significantly higher than that of digital angiography (3.8 ± 0.9, p = 0.011) and equivalent to that of DSA (4.3 ± 0.8, p = 0.651).ConclusionsRadiation exposure during AVS can be reduced by approximately half for both patients and operators by using DTDA and spot fluoroscopy without sacrificing image quality.

  20. Evaluation of Glucose-6-Phosphate Dehydrogenase stability in stored blood samples.

    Science.gov (United States)

    Jalil, Norunaluwar; Azma, Raja Zahratul; Mohamed, Emida; Ithnin, Azlin; Alauddin, Hafiza; Baya, Siti Noor; Othman, Ainoon

    2016-01-01

    Glucose-6-Phosphate Dehydrogenase (G6PD) deficiency is the commonest cause of neonatal jaundice in Malaysia. Recently, OSMMR2000-D G6PD Assay Kit has been introduced to quantitate the level of G6PD activity in newborns delivered in Universiti Kebangsaan Malaysia Medical Centre (UKMMC). As duration of sample storage prior to analysis is one of the matters of concern, this study was conducted to identify the stability of G6PD enzyme during storage. A total of 188 cord blood samples from normal term newborns delivered at UKMMC were selected for this study. The cord bloods samples were collected in ethylene-diamine-tetra-acetic acid (EDTA) tubes and refrigerated at 2-8 °C. In addition, 32 out of 188 cord blood samples were spotted on chromatography paper, air-dried and stored at room temperature. G6PD enzyme activities were measured daily for 7 days using the OSMMR2000-D G6PD Assay Kit on both the EDTA blood and dried blood samples. The mean value for G6PD activity was compared between days of analysis using Student Paired T-Test. In this study, 172 out of 188 cord blood samples showed normal enzyme levels while 16 had levels corresponding to severe enzyme deficiency. The daily mean G6PD activity for EDTA blood samples of newborns with normal G6PD activity showed a significant drop on the fourth day of storage (p samples with severely deficient G6PD activity, significant drop was seen on third day of storage (p = 0.002). Analysis of dried cord blood showed a significant reduction in enzyme activity as early as the second day of storage (p = 0.001). It was also noted that mean G6PD activity for spotted blood samples were lower compared to those in EDTA tubes for all days (p = 0.001). Thus, EDTA blood samples stored at 2-8 °C appeared to have better stability in terms of their G6PD enzyme level as compared to dried blood samples on filter paper, giving a storage time of up to 3 days.

  1. Correlation between Sweet Spots of Glycopeptides and Polymorphism of the Matrix Crystal in MALDI Samples.

    Science.gov (United States)

    Nishikaze, Takashi; Okumura, Hisako; Jinmei, Hiroshi; Amano, Junko

    2012-01-01

    A standard dried-droplet preparation using 2,5-dihydroxybenzoic acid (2,5-DHBA) as the matrix results in a large variation in signal intensity and poor shot-to-shot reproducibility in matrix-assisted laser desorption/ionization (MALDI). We expected that the differences can be attributed to the nature of the crystal structures in the region of the "sweet spot" within the MALDI samples. 2,5-DHBA crystals with and without analytes on a target plate obtained by means of a dried-droplet preparation contain two polymorphs, which can be distinguished by Raman spectra. In comparing the Raman image with the MS image, a clear correlation between the signal distribution of glycopeptides and hydrophilic peptides and the specific crystal form of 2,5-DHBA could be made. The ionization of hydrophobic peptides appears to proceed in both types of polymorphic crystals. In addition, the derivatization of glycopeptides with a pyrene group enabled us to detect glycopeptides regardless the crystal form. As the result, the number of sweet spots increased and MS spectra with a high signal intensity were obtained. The results suggest that the introduction of a hydrophobic/aromatic moiety to glycopeptides results in a more successful MALDI analysis due to the effective incorporation of the analyte into matrix crystals.

  2. A study of correlations between crude oil spot and futures markets: A rolling sample test

    Science.gov (United States)

    Liu, Li; Wan, Jieqiu

    2011-10-01

    In this article, we investigate the asymmetries of exceedance correlations and cross-correlations between West Texas Intermediate (WTI) spot and futures markets. First, employing the test statistic proposed by Hong et al. [Asymmetries in stock returns: statistical tests and economic evaluation, Review of Financial Studies 20 (2007) 1547-1581], we find that the exceedance correlations were overall symmetric. However, the results from rolling windows show that some occasional events could induce the significant asymmetries of the exceedance correlations. Second, employing the test statistic proposed by Podobnik et al. [Quantifying cross-correlations using local and global detrending approaches, European Physics Journal B 71 (2009) 243-250], we find that the cross-correlations were significant even for large lagged orders. Using the detrended cross-correlation analysis proposed by Podobnik and Stanley [Detrended cross-correlation analysis: a new method for analyzing two nonstationary time series, Physics Review Letters 100 (2008) 084102], we find that the cross-correlations were weakly persistent and were stronger between spot and futures contract with larger maturity. Our results from rolling sample test also show the apparent effects of the exogenous events. Additionally, we have some relevant discussions on the obtained evidence.

  3. Intensive Blood Pressure Reduction and Spot Sign in Intracerebral Hemorrhage: A Secondary Analysis of a Randomized Clinical Trial.

    Science.gov (United States)

    Morotti, Andrea; Brouwers, H Bart; Romero, Javier M; Jessel, Michael J; Vashkevich, Anastasia; Schwab, Kristin; Afzal, Mohammad Rauf; Cassarly, Christy; Greenberg, Steven M; Martin, Renee Hebert; Qureshi, Adnan I; Rosand, Jonathan; Goldstein, Joshua N

    2017-08-01

    The computed tomographic angiography (CTA) spot sign is associated with intracerebral hemorrhage (ICH) expansion and may mark those patients most likely to benefit from intensive blood pressure (BP) reduction. To investigate whether the spot sign is associated with ICH expansion across a wide range of centers and whether intensive BP reduction decreases hematoma expansion and improves outcome in patients with ICH and a spot sign. SCORE-IT (Spot Sign Score in Restricting ICH Growth) is a preplanned prospective observational study nested in the Antihypertensive Treatment of Acute Cerebral Hemorrhage II (ATACH-II) randomized clinical trial. Participants included consecutive patients with primary ICH who underwent a CTA within 8 hours from onset at 59 sites from May 15, 2011, through December 19, 2015. Data were analyzed for the present study from July 1 to August 31, 2016. Patients in ATACH-II were randomized to intensive (systolic BP target, sign, and 24 of 123 without missing data (19.5%) experienced ICH expansion. The spot sign was associated with expansion with sensitivity of 0.54 (95% CI, 0.34-0.74) and specificity of 0.63 (95% CI, 0.53-0.72). After adjustment for potential confounders, intensive BP treatment was not associated with a significant reduction of ICH expansion (relative risk, 0.83; 95% CI, 0.27-2.51; P = .74) or improved outcome (relative risk of 90-day modified Rankin Scale score ≥4, 1.24; 95% CI, 0.53-2.91; P = .62) in spot sign-positive patients. The predictive performance of the spot sign for ICH expansion was lower than in prior reports from single-center studies. No evidence suggested that patients with ICH and a spot sign specifically benefit from intensive BP reduction. clinicaltrials.gov Identifier: NCT01176565.

  4. Dried blood spots, valid screening for viral hepatitis and human immunodeficiency virus in real-life

    DEFF Research Database (Denmark)

    Mössner, Belinda K; Staugaard, Benjamin; Jensen, Janne

    2016-01-01

    to a combined nucleic acid test (NAT) for HBV DNA, HCV RNA and HIV RNA. RESULTS: Samples from 404 subjects were screened (85 CHB, 116 CHC, 114 HIV and 99 blood donors). DBS had a sensitivity of > 96% and a specificity of > 98% for the detection of all three infections. NAT testing did not improve sensitivity...

  5. High-performance liquid chromatography determination of dapsone, monoacetyldapsone, and pyrimethamine in filter paper blood spots

    DEFF Research Database (Denmark)

    Rønn, A M; Lemnge, M M; Angelo, H R

    1995-01-01

    A high-performance liquid chromatography method for the simultaneous analysis of dapsone (DDS), the major metabolite of DDS, monoacetyldapsone (MADDS), and pyrimethamine (PYR) was modified for capillary blood samples obtained by finger prick and dried on filter paper. Limit of quantitation using...

  6. Estimation of 24-hour urinary sodium excretion using spot urine samples.

    Science.gov (United States)

    Rhee, Moo-Yong; Kim, Ji-Hyun; Shin, Sung-Joon; Gu, Namyi; Nah, Deuk-Young; Hong, Kyung-Soon; Cho, Eun-Joo; Sung, Ki-Chul

    2014-06-20

    The present study evaluated the reliability of equations using spot urine (SU) samples in the estimation of 24-hour urine sodium excretion (24-HUNa). Equations estimating 24-HUNa from SU samples were derived from first-morning SU of 101 participants (52.4 ± 11.1 years, range 24-70 years). Equations developed by us and other investigators were validated with SU samples from a separate group of participants (n = 224, 51.0 ± 10.9 years, range 24-70 years). Linear, quadratic, and cubic equations were derived from first-morning SU samples because these samples had a sodium/creatinine ratio having the highest correlation coefficient for 24-HUNa/creatinine ratio (r = 0.728, p quadratic, and cubic equations developed from our study were not significantly different from measured 24-HUNa, while estimated 24-HUNa by previously developed equations were significantly different from measured 24-HUNa values. The limits of agreement between measured and estimated 24-HUNa by six equations exceeded 100 mmol/24-hour in the Bland-Altman analysis. All equations showed a tendency of under- or over-estimation of 24-HUNa, depending on the level of measured 24-HUNa. Estimation of 24-HUNa from single SU by equations as tested in the present study was found to be inadequate for the estimation of an individual's 24-HUNa.

  7. Multidrug resistant Salmonellae isolated from blood culture samples ...

    African Journals Online (AJOL)

    This study investigates the prevalence of R-plasmids in Salmonella sp. isolated from blood samples of suspected typhoid patients in Warri, Nigeria. A total of 136 blood samples were collected between May and December,2009 and screened for the presence of Salmonellae using standard blood culture techniques of which ...

  8. Measurements of purine derivatives and creatinine in spot urine samples of Chinese yellow cattle

    International Nuclear Information System (INIS)

    Xing, Z.; Xi, W.B.; Mo, F.; Liu, J.W.; Yang, Y.F.; Chen, X.B.

    2004-01-01

    An experiment was conducted using 18 Chinese Yellow Cattle located in 5 farms to study how supplementation of fermentable energy to low quality straw-based rations would improve rumen microbial protein synthesis. Within each farm, the animals were fed on five diets. Diets 1-2 were typical rice straw + by-products used by farmers and were low in fermentable energy content; Diets 3- 5 were more balanced, containing a higher content of fermentable energy. Purine derivatives (PD) and creatinine in spot urine samples were measured. The results showed that the PD to creatinine ratio was significantly higher with Diets 3-5 than with Diets 1-2. Organic matter digestibility and thus organic matter intake was also higher with Diets 3-5 compared to Diets 1-2. The results indicted that the efficiency of microbial protein synthesis could be improved by balancing the diet. (author)

  9. Quantification of 2-methylcitric acid in dried blood spots improves newborn screening for propionic and methylmalonic acidemias.

    Science.gov (United States)

    Al-Dirbashi, Osama Y; McIntosh, Nathan; Chakraborty, Pranesh

    2017-06-01

    Background Newborn screening for propionic acidemia and methylmalonic acidurias using the marker propionylcarnitine (C3) is neither sensitive nor specific. Using C3 to acetylcarnitine (C3/C2) ratio, together with conservative C3 cut-offs, can improve screening sensitivity, but the false positive rate remains high. Incorporating the marker 2-methylcitric acid has been suggested, to improve the positive predictive value for these disorders without compromising the sensitivity. Methods Between July 2011 and December 2012 at the Newborn Screening Ontario laboratory, all neonatal dried blood spot samples that were reported as screen positive for propionic acidemia or methylmalonic acidurias based on elevated C3 and C3/C2 ratio were analyzed for 2-methylcitric acid, using liquid chromatography tandem mass spectrometry. Results Of 222,420 samples screened, 103 were positive for methylmalonic acidurias or propionic acidemia using C3 and C3/C2 ratio as markers. There were nine true positives: propionic acidemia (n = 3), Cobalamin (Cbl) A (n=1), and Cbl C (n = 5). Among false positives there were 72 neonates not affected, 20 with maternal B 12 deficiency, and two incidental finding (transcobalamin II and unclassified Cbl defect). 2-Methylcitric acid was analyzed in all 103 samples and ranged between 0.1 and 89.4 µmol/l (reference range 0.04-0.36). Only 14 samples exceeded the set 2-methylcitric acid cut-off of 1.0 µmol/l, including the samples from all nine true positives. Conclusion By including 2-methylcitric acid in the screening algorithm, the positive predictive value of our primary and secondary screening targets improved from 8.7 to 64.3%. This would have eliminated 89 unnecessary referrals while maintaining 100% sensitivity.

  10. Immune trypanolysis test with blood spotted on filter paper for epidemiological surveillance of sleeping sickness.

    Science.gov (United States)

    Camara, Oumou; Camara, Mamadou; Lejon, Veerle; Ilboudo, Hamidou; Sakande, Hassane; Léno, Mamadou; Büscher, Philippe; Bucheton, Bruno; Jamonneau, Vincent

    2014-07-01

    The immune trypanolysis test (TL) is an accurate sero-diagnostic tool increasingly implemented for sleeping sickness medical surveillance, but it is restricted to the reference laboratories. To facilitate storage and transport of the test specimen, we developed a protocol for the examination of blood spotted on filter paper (TL-fp) that can be stored and shipped at ambient temperature. We compared its performance with the classical TL on plasma (TL-pl) that needs to be kept frozen until use. The study was conducted in active foci of the Republic of Guinea. In total, 438 specimens from treated and untreated sleeping sickness patients and serological suspects were tested with both methods. TL-fp gave significantly less positive results than TL-pl, but all the confirmed sleeping sickness cases were positive with the TL-fp protocol. TL-fp appears to offer a good compromise between feasibility and sensitivity to detect currently infected subjects who play a role in the transmission of Trypanosoma brucei gambiense and is useful for contributing to the elimination of gambiense sleeping sickness. © 2014 John Wiley & Sons Ltd.

  11. Quantification of DNA in Neonatal Dried Blood Spots by Adenine Tandem Mass Spectrometry.

    Science.gov (United States)

    Durie, Danielle; Yeh, Ed; McIntosh, Nathan; Fisher, Lawrence; Bulman, Dennis E; Birnboim, H Chaim; Chakraborty, Pranesh; Al-Dirbashi, Osama Y

    2018-01-02

    Newborn screening programs have expanded to include molecular-based assays as first-tier tests and the success of these assays depends on the quality and yield of DNA extracted from neonatal dried blood spots (DBS). To meet high throughput and rapid turnaround time requirements, newborn screening laboratories adopted rapid DNA extraction methods that produce crude extracts. Quantification of DNA in neonatal DBS is not routinely performed due to technical challenges; however, this may enhance the performance of assays that are sensitive to amounts of input DNA. In this study, we developed a novel high throughput method to quantify total DNA in DBS. It is based on specific acid-catalyzed depurination of DNA followed by mass spectrometric quantification of adenine. The amount of adenine was used to calculate DNA quantity per 3.2 mm DBS. Reference intervals were established using archived, neonatal DBS (n = 501) and a median of 130.6 ng of DNA per DBS was obtained, which is in agreement with literature values. The intra- and interday variations were quantification were 12.5 and 37.8 nmol/L adenine, respectively. We demonstrated that DNA from neonatal DBS can be successfully quantified in high throughput settings using instruments currently deployed in NBS laboratories.

  12. Dried Blood Spots for Measuring Vibrio cholerae-specific Immune Responses.

    Directory of Open Access Journals (Sweden)

    Anita S Iyer

    2018-01-01

    Full Text Available Vibrio cholerae causes over 2 million cases of cholera and 90,000 deaths each year. Serosurveillance can be a useful tool for estimating the intensity of cholera transmission and prioritizing populations for cholera control interventions. Current methods involving venous blood draws and downstream specimen storage and transport methods pose logistical challenges in most settings where cholera strikes. To overcome these challenges, we developed methods for determining cholera-specific immune responses from dried blood spots (DBS.As conventional vibriocidal assay methods were unsuitable for DBS eluates from filter paper, we adopted a drop-plate culture method. We show that DBS collected from volunteers in South Sudan, and stored for prolonged periods in field conditions, retained functional vibriocidal antibodies, the titers of which correlated with paired serum titers determined by conventional spectrophotometric methods (r = 0.94, p = 0.00012. We also showed that eluates from DBS Serum Separator cards could be used with conventional spectrophotometric vibriocidal methods, and that they correlated with paired serum at a wide range of titers (r = 0.96, p<0.0001. Similarly, we used ELISA methods to show that V. cholerae O-specific polysaccharide antibody responses from DBS eluates correlated with results from paired serum for IgG (r = 0.85, p = 0.00006, IgM (r = 0.79, p = 0.00049 and IgA (r = 0.73, p = 0.0019, highlighting its potential for use in determination of isotype-specific responses. Storage of DBS cards at a range of temperatures did not change antibody responses.In conclusion, we have developed and demonstrated a proof-of-concept for assays utilizing DBS for assessing cholera-specific immune responses.

  13. Blood samples in the neutron beam

    International Nuclear Information System (INIS)

    Anon.

    2012-01-01

    Whether crocodile, platypus, man, or chicken: Always the hemoglobin in the red blood cells has the same task. It carries oxygen from the lungs throughout the body. In investigative manner and international team around Dr. Andreas Stadler from the Julic Research Center has deciphered in detail, how and why the hemoglobines of these creatures nevertheless differ. Their findings are among others interesting for the research on artificial blood.

  14. Combination syringe provides air-free blood samples

    Science.gov (United States)

    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.

  15. Usefulness of Dried Blood Spots (DBS) to perform hepatitis C virus genotyping in drug users in Senegal.

    Science.gov (United States)

    Ndiaye, O; Gozlan, J; Diop-Ndiaye, H; Sall, A S; Chapelain, S; Leprêtre, A; Maynart, M; Gueye, M; Lo, G; Thiam, M; Ba, I; Lacombe, K; Girard, P M; Mboup, S; Kane, C T

    2017-03-01

    The aim of this pilot study was to analyze the Hepatitis C Virus (HCV) genotypes circulating in Senegal among Drug User (DUs), using Dried Blood Spots (DBS) as RNA source for molecular assays. Heroin and/or cocaine users (n = 506) were recruited in Dakar from April to July 2011, using a Respondent Driven Sampling (RDS) method. DBS preparation consisted of five drops of whole blood from finger applied to a Whatman paper card. HCV infection was screened by the detection of anti-HCV antibodies, using a rapid immune-chromatographic test. HCV RNA was quantified on anti-HCV positive DBS, using the Abbott RealTime HCV® Genotyping was performed on DBS with detectable viral load with Versant® HCV Genotype 2.0 Assay (LiPA) and Abbott RealTime HCV Genotype II assay®. Among the 506 participants, 120 were tested as positive for anti-HCV antibodies and their samples were analyzed for HCV RNA viral load and genotype. Out of the 120 DBS tested, HCV RNA was detected on 25 (20.8%). The median viral load was 15,058 IU/ml (ranging from 710 to 766,740 IU/ml). All positive DBS were suitable for the genotyping assay, that showed a predominance of genotype 1 (21/25) including 16 genotypes 1a and 5 genotypes 1b. HCV genotype 1 prevails in a DU population in Dakar. DBS could be useful for HCV RNA genotyping, but optimal storage conditions should required avoiding RNA impairment. Acknowledging this limitation, DBS could be a great interest for detecting and genotyping HCV viremic patients. J. Med. Virol. 89:484-488, 2017. © 2015 Wiley Periodicals, Inc. © 2015 Wiley Periodicals, Inc.

  16. A novel tandem mass spectrometry method for first-line screening of mainly beta-thalassemia from dried blood spots.

    Science.gov (United States)

    Yu, Chaowen; Huang, Shuodan; Wang, Ming; Zhang, Juan; Liu, Hao; Yuan, Zhaojian; Wang, Xingbin; He, Xiaoyan; Wang, Jie; Zou, Lin

    2017-02-10

    Traditional methods for thalassemia screening are time-consuming and easily affected by cell hemolysis or hemoglobin degradation in stored blood samples. Tandem mass spectrometry (MS/MS) proved to be an effective technology for sickle cell disorders (SCD) screening. Here, we developed a novel MS/MS method for β-thalassemia screening from dried blood spots (DBS). Stable isotopic-labeled peptides were used as internal standards for quantification and calculation of the α:β-globin ratios. We used the α:β-globin ratio cutoffs to differentiate between normal individuals and patients with thalassemia. About 781 patients and 300 normal individuals were analyzed. The α:β-globin ratios showed significant difference between normal and β-thalassemia patients (Pthalassemia mutation. In the parallel study, all cases screened for suspected thalassemia from six hundred DBS samples by using this MS/MS method were successfully confirmed by genotyping. The intra-assay and inter-assay CVs of the ratios ranged from 2.4% to 3.9% and 4.7% to 7.1%, and there was no significant sample carryover or matrix effect for this MS/MS method. Combined with SCD screening, this MS/MS method could be used as a first-line screening assay for both structural and expression abnormalities of human hemoglobin. Traditional methods for thalassemia screening were depending on the structural integrity of tetramers and could be affected by hemolysis and degradation of whole blood samples, especially when stored. We used proteospecific peptides produced by the tryptic digestion of each globin to evaluate the production ratio between α- and β-globin chains, which turned out to be quite stable even when stored for more than two months. Though most of the peptides were specific to α-globin or β-globin, we only chose four most informative peptides and its stable isotopic-labeled peptides as internal standards for analysis, which could obtain a high accuracy. Currently, we are the first to address the

  17. Dried blood spot measurement of pregnancy-associated plasma protein A (PAPP-A) and free beta-subunit of human chorionic gonadotropin (beta-hCG) from a low-resource setting

    NARCIS (Netherlands)

    Browne, J. L.; Schielen, P. C. J. I.; Belmouden, I.; Pennings, J. L. A.; Klipstein-Grobusch, K.

    ObjectivesThe objectives of the article is to compare pregnancy-associated plasma protein A (PAPP-A) and free -subunit of human chorionic gonadotropin (-hCG) concentrations in dried blood spots (DBSs) with serum of samples obtained from a public hospital in a low-resource setting and to evaluate

  18. Validation of a Multiplex Tandem Mass Spectrometry Method for the Detection of Selected Lysosomal Storage Diseases in Dried Blood Spots

    Directory of Open Access Journals (Sweden)

    Graziela Schmitt Ribas PhD

    2017-02-01

    Full Text Available Background: Interest in screening methods for lysosomal storage diseases (LSDs has increased in recent years, since early diagnosis and treatment are essential to prevent or attenuate the onset of symptoms and the complications of these diseases. In the current work, we evaluated the performance of tandem mass spectrometry (MS/MS for the detection of some LSDs, aiming the future use of this methodology for the screening of these disorders. Methods: Standard curves and quality control dried blood spots were assayed to evaluate the precision, linearity, and accuracy. A total of 150 controls were grouped according to age and subjected to measurement of lysosomal enzymes deficient in Niemann-Pick A/B, Krabbe, Gaucher, Fabry, Pompe, and Mucopolysaccharidosis type I diseases. Samples from 59 affected patients with a diagnosis of LSDs previously confirmed by fluorimetric methods were analyzed. Results: Data from standard calibration demonstrated good linearity and accuracy and the intra- and interassay precisions varied from 1.17% to 11.60% and 5.39% to 31.24%, respectively. Except for galactocerebrosidase and α- l -iduronidase, enzyme activities were significantly higher in newborns compared to children and adult controls. Affected patients presented enzymatic activities significantly lower compared to all control participants. Conclusion: Our results show that MS/MS is a promising methodology, suitable for the screening of LSDs, but accurate diagnoses will depend on its correlation with other biochemical and/or molecular analyses.

  19. Studies of U in the blood of two population samples

    International Nuclear Information System (INIS)

    Segovia, N.; Olguin, M.E.; Romero, M.

    1986-01-01

    The present work, attempts to establish the statistical distribution of blood uranium in a population of the same community, similar in age and in living patterns. U traces were evaluated by a fission track technique both in whole blood and plasma samples. Dried samples were compressed into pellets and irradiated in a nuclear reactor using the external detector method. For U quantification, standard U samples were used. A comparative sampling of U content in blood samples from a group of radiation exposed workers and another of leukemia patients was also carried out. Results from the sampling groups are reported and discussed. (author)

  20. High explosive spot test analyses of samples from Operable Unit (OU) 1111

    Energy Technology Data Exchange (ETDEWEB)

    McRae, D.; Haywood, W.; Powell, J.; Harris, B.

    1995-01-01

    A preliminary evaluation has been completed of environmental contaminants at selected sites within the Group DX-10 (formally Group M-7) area. Soil samples taken from specific locations at this detonator facility were analyzed for harmful metals and screened for explosives. A sanitary outflow, a burn pit, a pentaerythritol tetranitrate (PETN) production outflow field, an active firing chamber, an inactive firing chamber, and a leach field were sampled. Energy dispersive x-ray fluorescence (EDXRF) was used to obtain semi-quantitative concentrations of metals in the soil. Two field spot-test kits for explosives were used to assess the presence of energetic materials in the soil and in items found at the areas tested. PETN is the major explosive in detonators manufactured and destroyed at Los Alamos. No measurable amounts of PETN or other explosives were detected in the soil, but items taken from the burn area and a high-energy explosive (HE)/chemical sump were contaminated. The concentrations of lead, mercury, and uranium are given.

  1. Technical Advances for the Clinical Genomic Evaluation of Sudden Cardiac Death: Verification of Next-Generation Sequencing Panels for Hereditary Cardiovascular Conditions Using Formalin-Fixed Paraffin-Embedded Tissues and Dried Blood Spots.

    Science.gov (United States)

    Baudhuin, Linnea M; Leduc, Charles; Train, Laura J; Avula, Rajeswari; Kluge, Michelle L; Kotzer, Katrina E; Lin, Peter T; Ackerman, Michael J; Maleszewski, Joseph J

    2017-12-01

    Postmortem genetic testing for heritable cardiovascular (CV) disorders is often lacking because ideal specimens (ie, whole blood) are not retained routinely at autopsy. Formalin-fixed paraffin-embedded tissue (FFPET) is ubiquitously collected at autopsy, but DNA quality hampers its use with traditional sequencing methods. Targeted next-generation sequencing may offer the ability to circumvent such limitations, but a method has not been previously described. The primary aim of this study was to develop and evaluate the use of FFPET for heritable CV disorders via next-generation sequencing. Nineteen FFPET (heart) and blood (whole blood or dried blood spot) specimens underwent targeted next-generation sequencing using a custom panel of 101 CV-associated genes. Nucleic acid yield and quality metrics were evaluated in relation to FFPET specimen age (6 months to 15 years; n=14) and specimen type (FFPET versus whole blood and dried blood spot; n=12). Four FFPET cases with a clinical phenotype of heritable CV disorder were analyzed. Accuracy and precision were 100% concordant between all sample types, with read depths >100× for most regions tested. Lower read depth, as low as 40×, was occasionally observed with FFPET and dried blood spot. High-quality DNA was obtained from FFPET samples as old as 15 years. Genomic analysis of FFPET from the 4 phenotype-positive/genotype unknown cases all revealed putative disease-causing variants. Similar performance characteristics were observed for next-generation sequencing of FFPET, whole blood, and dried blood spot in the evaluation of inherited CV disorders. Although blood is preferable for genetic analyses, this study offers an alternative when only FFPET is available. © 2017 American Heart Association, Inc.

  2. PCR-based pooling of dried blood spots for detection of malaria parasites: optimization and application to a cohort of Ugandan children.

    Science.gov (United States)

    Hsiang, Michelle S; Lin, Michael; Dokomajilar, Christian; Kemere, Jordan; Pilcher, Christopher D; Dorsey, Grant; Greenhouse, Bryan

    2010-10-01

    Sensitive, high-throughput methods to detect malaria parasites in low-transmission settings are needed. PCR-based pooling strategies may offer a solution. We first used laboratory-prepared samples to compare 2 DNA extraction and 4 PCR detection methods across a range of pool sizes and parasite densities. Pooled Chelex extraction of DNA, followed by nested PCR of cytochrome b, was the optimal strategy, allowing reliable detection of a single low-parasitemic sample (100 parasites/μl) in pool sizes up to 50. This PCR-based pooling strategy was then compared with microscopy using 891 dried blood spots from a cohort of 77 Ugandan children followed for 2 years in an urban setting of low endemicity. Among 419 febrile episodes, 35 cases of malaria were detected using the PCR-based pooling strategy and 40 cases using microscopy. All five cases of malaria not detected by PCR were from samples stored for >2 years with parasitemia of parasites using dried blood spots offers a sensitive and efficient approach for malaria surveillance in low-transmission settings, enabling improved detection of asymptomatic submicroscopic infections and dramatic savings in labor and costs.

  3. Dried Blood Spot Reference Intervals for Steroids and Amino Acids in a Neonatal Cohort of the National Children's Study.

    Science.gov (United States)

    Dietzen, Dennis J; Bennett, Michael J; Lo, Stanley F; Grey, Vijay L; Jones, Patti M

    2016-12-01

    Reference intervals from children are limited by access to healthy children and their limited blood volumes. In this study we set out to fill gaps in pediatric reference intervals for amino acids and steroid hormones using dried blood spots (DBS) from a cohort of the National Children's Study. Deidentified DBS annotated with age, birthweight, sex, and geographic location were obtained from 310 newborns aged 0-4 days and analyzed for 25 amino acids and 4 steroid hormones using LC-MS/MS. Nonparametric statistical approaches were used to generate the 2.5th-97.5th percentile distributions for newborns. Paired plasma/DBS specimens were used to mathematically transform DBS reference intervals to corresponding plasma intervals. 10 of 25 DBS amino acid distributions were dependent on sex. There was little correlation with age, birthweight, or geographic location over the first 4 days of life. In most cases, transformation of DBS distributions to plasma distributions faithfully reflected independent studies of newborn plasma amino acid distributions. In general newborn steroid distributions were negatively correlated with age and birthweight over the first 4 days of life. Data distributions for the 4 steroids were not found related to geographic location, but testosterone concentrations displayed sex dependence. Transformation of DBS distributions to plasma intervals did not faithfully replicate other neonate steroid reference intervals determined directly with plasma. These data demonstrate the feasibility and utility of deriving newborn reference intervals from large numbers of archived DBS samples such as those obtained from the National Children's Study biobank. © 2016 American Association for Clinical Chemistry.

  4. Congenital cytomegalovirus: association between dried blood spot viral load and hearing loss.

    Science.gov (United States)

    Walter, S; Atkinson, C; Sharland, M; Rice, P; Raglan, E; Emery, V C; Griffiths, P D

    2008-07-01

    To investigate the relation between cytomegalovirus (CMV) viral load on dried blood spots (DBS) from newborn biochemical screening ("Guthrie") cards, and sensorineural hearing loss (SNHL) in congenital CMV. Cross-sectional study with retrospective case-note review. Seven paediatric audiology departments in the United Kingdom. 84 children, median age 7 years: 43 with known congenital CMV, 41 with unexplained SNHL. Half a DBS was tested for CMV DNA viral load by quantitative real-time polymerase chain reaction (PCR). Pure tone average hearing thresholds (0.5-4 kHz). DBS CMV DNA viral load significantly correlated with hearing thresholds for the worse and better hearing ears (Spearman's rank correlations: r = 0.445, p = 0.008 and r = 0.481, p = 0.004 respectively). Multivariable logistic regression showed that the effect of DBS viral load on the risk of SNHL remained important, when age and central nervous system involvement had been taken into account (odds ratio (OR) 2.76, 95% confidence interval (CI) 1.14 to 6.63, p = 0.024). The mean log DBS viral load was significantly higher in children with SNHL than in those with normal hearing (2.69 versus 1.64, 95% CI -1.84 to -0.27, p = 0.01). 8/35 (23%) children with unexplained SNHL tested positive for CMV DNA on DBS. One false positive result was obtained. The risk of SNHL increased with DBS viral load. Further studies should investigate whether DBS CMV testing has a role in identifying asymptomatic congenitally infected neonates at risk of SNHL, and whether antiviral treatment can reduce this risk.

  5. The stability of hexacosanoyl lysophosphatidylcholine in dried-blood spot quality control materials for X-linked adrenoleukodystrophy newborn screening.

    Science.gov (United States)

    Haynes, Christopher A; De Jesús, Víctor R

    2015-01-01

    Newborn screening for X-linked adrenoleukodystrophy utilizes tandem mass spectrometry to analyze dried-blood spot specimens. Quality control materials (dried-blood spots enriched with hexacosanoyl lysophosphatidylcholine) were prepared and stored at different temperatures for up to 518days to evaluate the stability of this biomarker for X-linked adrenoleukodystrophy. Dried-blood spot storage included desiccant (45, 171, and 518days) or omitted desiccant (53days at >90% relative humidity). Specimens were stored for 171 and 518days at -20°C, 4°C, ambient temperature, and 37°C. Each weekday for 45days, a bag of specimens stored at 4°C was warmed to ambient temperature and one specimen was removed for storage at -80°C. Specimens were analyzed by high-performance liquid-chromatography electrospray ionization tandem mass spectrometry and data was plotted as concentration (micromoles per liter) vs. time. Linear regression provided slope and y-intercept values for each storage condition. Small slope values (0.01 or less) and y-intercept values close to the enrichment indicated less than 11% loss of hexacosanoyl lysophosphatidylcholine under all storage conditions tested. Quality control materials for X-linked adrenoleukodystrophy are stable for at least 1year when stored with desiccant. Published by Elsevier Inc.

  6. The pathology of facial vein blood sampling in mice

    DEFF Research Database (Denmark)

    Hansen, Ket; Harslund, Jakob le Fèvre; Bollen, Peter

    2014-01-01

    Introduction: The use of retro-orbital blood sampling is prohibited in Denmark. For this reason, alternative methods are used for obtaining larger blood samples of a good quality. The facial vein is generally recommended for this. However, we have experienced discomfort for mice subjected to facial...... vein blood sampling. Therefore, we investigated if this technique was associated with pathological changes of the jaw region. Methods: 43 NMRI mice were subjected to facial vein blood sampling by using the lancet method during 12 months, starting at the age of 8 weeks. The mice were restrained manually...... by the scruff and a lancet was placed 2-3 mm caudally to the freckle on the lower jaw, and the skin was punctured. After sampling, brief compression by a cotton swab was applied, if bleeding did not stop. Two days after the last blood sampling, the mice were euthanized by an overdose of pentobarbital...

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

    Science.gov (United States)

    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.

  8. DETERMINATION OF REFERENCE VALUES FOR TREC AND KREC IN DRY BLOOD SPOTS OF NEWBORNS FROM DIFFERENT GESTATION AGES IN SVERDLOVSK REGION

    Directory of Open Access Journals (Sweden)

    S. S. Deryabina

    2018-01-01

    Full Text Available As a preparatory stage for implementation of genetic testing for severe combined immunodeficiency under a neonatal screening program, a study was performed in Sverdlovsk Region which concerned quantitative determination of T and B cell neogenesis markers (TREC and KREC, respectively in blood of conditionally healthy newborns. Archived samples of dry blood spots collected in test-forms for routine neonatal screening were used as biological material for the study of full-term 26 girls and 26 boys who did not exhibit serious illnesses during first year of their life. In addition, we investigated potential effects of foetal gestational age upon the number of TREC and KREC in preterm infants. Blood samples from 55 preterm infants (23 to 36 gestational weeks were also examined. It was shown that the levels of TREC and KREC increased sequentially with the increased gestation terms, but the quantitative changes of markers showed different dynamics. In this respect, the recommended terms of blood sample collection for SCID screening is entirely consistent with timing of blood sampling for routine newborn screening. An alternative result was obtained with a complete absence of TREC or KREC in blood sample of a newborn, irrespectively of prematurity degree (at valid copy numbers of a control gene which should serve as an indication for immediate consulting of the child by immunologist and in-depth immunological examination, because it may be a first prognostic sign of a fatal disease. In order to obtain correct cut-off levels for TREC/KREC, additional studies are needed on a larger sample of newborns (1.000 to 5.000, followed by validation of the obtained reference boundaries in studies involving patients with different forms of primary immunodeficiencies. 

  9. Neonatal blood gas sampling methods | Goenka | South African ...

    African Journals Online (AJOL)

    Blood gas sampling is part of everyday practice in the care of babies admitted to the neonatal intensive care unit, particularly for those receiving respiratory support. There is little published guidance that systematically evaluates the different methods of neonatal blood gas sampling, where each method has its individual ...

  10. Diagnosis of immunodeficiency caused by a purine nucleoside phosphorylase defect by using tandem mass spectrometry on dried blood spots.

    Science.gov (United States)

    la Marca, Giancarlo; Canessa, Clementina; Giocaliere, Elisa; Romano, Francesca; Malvagia, Sabrina; Funghini, Silvia; Moriondo, Maria; Valleriani, Claudia; Lippi, Francesca; Ombrone, Daniela; Della Bona, Maria Luisa; Speckmann, Carsten; Borte, Stephan; Brodszki, Nicholas; Gennery, Andrew R; Weinacht, Katja; Celmeli, Fatih; Pagel, Julia; de Martino, Maurizio; Guerrini, Renzo; Wittkowski, Helmut; Santisteban, Ines; Bali, Pawan; Ikinciogullari, Aydan; Hershfield, Michael; Notarangelo, Luigi D; Resti, Massimo; Azzari, Chiara

    2014-07-01

    Purine nucleoside phosphorylase (PNP) deficiency is a rare form of autosomal recessive combined primary immunodeficiency caused by a enzyme defect leading to the accumulation of inosine, 2'-deoxy-inosine (dIno), guanosine, and 2'-deoxy-guanosine (dGuo) in all cells, especially lymphocytes. Treatments are available and curative for PNP deficiency, but their efficacy depends on the early approach. PNP-combined immunodeficiency complies with the criteria for inclusion in a newborn screening program. This study evaluate whether mass spectrometry can identify metabolite abnormalities in dried blood spots (DBSs) from affected patients, with the final goal of individuating the disease at birth during routine newborn screening. DBS samples from 9 patients with genetically confirmed PNP-combined immunodeficiency, 10,000 DBS samples from healthy newborns, and 240 DBSs from healthy donors of different age ranges were examined. Inosine, dIno, guanosine, and dGuo were tested by using tandem mass spectrometry (TMS). T-cell receptor excision circle (TREC) and kappa-deleting recombination excision circle (KREC) levels were evaluated by using quantitative RT-PCR only for the 2 patients (patients 8 and 9) whose neonatal DBSs were available. Mean levels of guanosine, inosine, dGuo, and dIno were 4.4, 133.3, 3.6, and 3.8 μmol/L, respectively, in affected patients. No indeterminate or false-positive results were found. In patient 8 TREC levels were borderline and KREC levels were abnormal; in patient 9 TRECs were undetectable, whereas KREC levels were normal. TMS is a valid method for diagnosis of PNP deficiency on DBSs of affected patients at a negligible cost. TMS identifies newborns with PNP deficiency, whereas TREC or KREC measurement alone can fail. Copyright © 2014 American Academy of Allergy, Asthma & Immunology. Published by Mosby, Inc. All rights reserved.

  11. Simultaneous Testing for 6 Lysosomal Storage Disorders and X-Adrenoleukodystrophy in Dried Blood Spots by Tandem Mass Spectrometry.

    Science.gov (United States)

    Tortorelli, Silvia; Turgeon, Coleman T; Gavrilov, Dimitar K; Oglesbee, Devin; Raymond, Kimiyo M; Rinaldo, Piero; Matern, Dietrich

    2016-09-01

    Newborn screening for lysosomal storage disorders (LSD) has revealed that late-onset variants of these conditions are unexpectedly frequent and therefore may evade diagnosis. We developed an efficient and cost-effective multiplex assay to diagnose six LSDs and several peroxisomal disorders in patients presenting with diverse phenotypes at any age. Three 3-mm dried blood spot (DBS) punches were placed into individual microtiter plates. One disc was treated with a cocktail containing acid sphingomyelinase-specific substrate and internal standard (IS). To the second DBS we added a cocktail containing substrate and IS for β-glucosidase, acid α-glucosidase, α-galactosidase A, galactocerebrosidase, and α-L-iduronidase. The third DBS was extracted with methanol containing d4-C26 lysophosphatidylcholine as IS and stored until the enzyme plates were combined and purified by liquid-liquid and solid-phase extraction. The extracts were evaporated, reconstituted with the extract from the lysophosphatidylcholine plate, and analyzed by flow injection tandem mass spectrometry. Reference intervals were determined by analysis of 550 samples from healthy controls. DBS from confirmed patients with 1 of the 6 LSDs (n = 33), X-adrenoleukodystrophy (n = 9), or a peroxisomal biogenesis disorder (n = 5), as well as carriers for Fabry disease (n = 17) and X-adrenoleukodystrophy (n = 5), were analyzed for assay validation. Prospective clinical testing of 578 samples revealed 25 patients affected with 1 of the detectable conditions. Our flow injection tandem mass spectrometry approach is amenable to high-throughput population screening for Hurler disease, Gaucher disease, Niemann-Pick A/B disease, Pompe disease, Krabbe disease, Fabry disease, X-adrenoleukodystrophy, and peroxisomal biogenesis disorder in DBS. © 2016 American Association for Clinical Chemistry.

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

    Science.gov (United States)

    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.

  13. Radioimmunoassay of ''free thyroxin'' in dried blood spots on filter paper - preliminary observations on the effective differentiation of subjects with congenital hypothyroidism from those with subnormal thyroxin-binding globulin and normal subjects

    International Nuclear Information System (INIS)

    Mizuta, H.; Miyai, K.; Ichihara, K.; Amino, N.; Harada, T.; Nose, O.; Tanizawa, O.

    1982-01-01

    In this sensitive, simple method for measuring ''free thyroxin'' (FT 4 ) in eluates of dried blood spots on filter paper by use of a radioimmunoassay kit (Amerlex Free T 4 RIA), the measurable range of FT 4 is 1.8 to 57 ng/L (equivalent to the concentration in serum), or 7 to 237 fg/tube. The mean coefficients of variation for within assay-within spots, within assay-between spots, and between assays were 5.3%, 5.0%, and 6.2%, respectively. FT 4 in blood spotted on filter paper is stable for at least a month when dried and kept at either -20 0 C, 4 0 C, room temperature (about 25 0 C), or 37 0 C. The results for FT 4 in dried blood spots correlated closely with the free-T 4 concentration in serum (r = 0.99). The method can be used to differentiate cases of primary and secondary hypothyroidism from normal subjects and those with subnormal thyroxin-binding globulin. This method may be useful in screening for congenital hypothyroidism, because sample-retesting is not necessary

  14. Dried blood spot testing for the antenatal screening of HTLV, HIV, syphilis, toxoplasmosis and hepatitis B and C: prevalence, accuracy and operational aspects

    Directory of Open Access Journals (Sweden)

    Ney Boa-Sorte

    2014-11-01

    Conclusions: The use of dried blood spot may represent a secure way to expedite access to results of vertically transmitted diseases in the prenatal period, particularly in regions with scarce healthcare resources.

  15. Exposure and risk factors to coxiella burnetii, spotted fever group and typhus group Rickettsiae, and Bartonella henselae among volunteer blood donors in Namibia.

    Directory of Open Access Journals (Sweden)

    Bruce H Noden

    Full Text Available The role of pathogen-mediated febrile illness in sub-Saharan Africa is receiving more attention, especially in Southern Africa where four countries (including Namibia are actively working to eliminate malaria. With a high concentration of livestock and high rates of companion animal ownership, the influence of zoonotic bacterial diseases as causes of febrile illness in Namibia remains unknown.The aim of the study was to evaluate exposure to Coxiella burnetii, spotted fever and typhus group rickettsiae, and Bartonella henselae using IFA and ELISA (IgG in serum collected from 319 volunteer blood donors identified by the Blood Transfusion Service of Namibia (NAMBTS. Serum samples were linked to a basic questionnaire to identify possible risk factors. The majority of the participants (64.8% had extensive exposure to rural areas or farms. Results indicated a C. burnetii prevalence of 26.1% (screening titre 1∶16, and prevalence rates of 11.9% and 14.9% (screening titre 1∶100 for spotted fever group and typhus group rickettsiae, respectively. There was a significant spatial association between C. burnetii exposure and place of residence in southern Namibia (P0.012, especially cattle (P>0.006, were also significantly associated with C. burnetii exposure. Males were significantly more likely than females to have been exposed to spotted fever (P<0.013 and typhus (P<0.011 group rickettsiae. Three (2.9% samples were positive for B. henselae possibly indicating low levels of exposure to a pathogen never reported in Namibia.These results indicate that Namibians are exposed to pathogenic fever-causing bacteria, most of which have flea or tick vectors/reservoirs. The epidemiology of febrile illnesses in Namibia needs further evaluation in order to develop comprehensive local diagnostic and treatment algorithms.

  16. A novel dried blood spot-LCMS method for the quantification of methotrexate polyglutamates as a potential marker for methotrexate use in children.

    Directory of Open Access Journals (Sweden)

    Ahmed F Hawwa

    Full Text Available Development and validation of a selective and sensitive LCMS method for the determination of methotrexate polyglutamates in dried blood spots (DBS.DBS samples [spiked or patient samples] were prepared by applying blood to Guthrie cards which was then dried at room temperature. The method utilised 6-mm disks punched from the DBS samples (equivalent to approximately 12 µl of whole blood. The simple treatment procedure was based on protein precipitation using perchloric acid followed by solid phase extraction using MAX cartridges. The extracted sample was chromatographed using a reversed phase system involving an Atlantis T3-C18 column (3 µm, 2.1 × 150 mm preceded by Atlantis guard column of matching chemistry. Analytes were subjected to LCMS analysis using positive electrospray ionization.The method was linear over the range 5-400 nmol/L. The limits of detection and quantification were 1.6 and 5 nmol/L for individual polyglutamates and 1.5 and 4.5 nmol/L for total polyglutamates, respectively. The method has been applied successfully to the determination of DBS finger-prick samples from 47 paediatric patients and results confirmed with concentrations measured in matched RBC samples using conventional HPLC-UV technique.The methodology has a potential for application in a range of clinical studies (e.g. pharmacokinetic evaluations or medication adherence assessment since it is minimally invasive and easy to perform, potentially allowing parents to take blood samples at home. The feasibility of using DBS sampling can be of major value for future clinical trials or clinical care in paediatric rheumatology.

  17. Dried Blood Spots Combined With Ultra-High-Performance Liquid Chromatography-Mass Spectrometry for the Quantification of the Antipsychotics Risperidone, Aripiprazole, Pipamperone, and Their Major Metabolites.

    Science.gov (United States)

    Tron, Camille; Kloosterboer, Sanne M; van der Nagel, Bart C H; Wijma, Rixt A; Dierckx, Bram; Dieleman, Gwen C; van Gelder, Teun; Koch, Birgit C P

    2017-08-01

    Risperidone, aripiprazole, and pipamperone are antipsychotic drugs frequently prescribed for the treatment of comorbid behavioral problems in children with autism spectrum disorders. Therapeutic drug monitoring (TDM) could be useful to decrease side effects and to improve patient outcome. Dried blood spot (DBS) sample collection seems to be an attractive technique to develop TDM of these drugs in a pediatric population. The aim of this work was to develop and validate a DBS assay suitable for TDM and home sampling. Risperidone, 9-OH risperidone, aripiprazole, dehydroaripiprazole, and pipamperone were extracted from DBS and analyzed by ultra-high-performance liquid chromatography-tandem mass spectrometry using a C18 reversed-phase column with a mobile phase consisting of ammonium acetate/formic acid in water or methanol. The suitability of DBS for TDM was assessed by studying the influence of specific parameters: extraction solution, EDTA carryover, hematocrit, punching location, spot volume, and hemolysis. The assay was validated with respect to conventional guidelines for bioanalytical methods. The method was linear, specific without any critical matrix effect, and with a mean recovery around 90%. Accuracy and imprecision were within the acceptance criteria in samples with hematocrit values from 30% to 45%. EDTA or hemolysis did not skew the results, and no punching carryover was observed. No significant influence of the spot volume or the punch location was observed. The antipsychotics were all stable in DBS stored 10 days at room temperature and 1 month at 4 or -80°C. The method was successfully applied to quantify the 3 antipsychotics and their metabolites in patient samples. A UHPLC-MS/MS method has been successfully validated for the simultaneous quantification of risperidone, 9-OH risperidone, aripiprazole, dehydroaripiprazole, and pipamperone in DBS. The assay provided good analytical performances for TDM and clinical research applications.

  18. 5C.07: A METHOD TO ESTIMATE 24-HOUR SODIUM EXCRETION THROUGH SPOT URINE SAMPLES AND ITS APPLICATION VALUE FOR TARGET-ORGAN DAMAGE ASSESSMENT.

    Science.gov (United States)

    Wang, H; Zhao, L; Xi, Y; Sun, N

    2015-06-01

    24-h urine sodium excretion is considered the most reliable method to evaluate the salt intakes. However, this method is cumbersome. So we want to develop formulas to estimate 24-h urinary sodium excretion using spot urinary samples in Chinese hypertensive population and explore the application value of this method in salt intake assessment and target organ damage. 1. We enrolled 510 cases of hospitalized patients with hypertension, 2/3 of them were arranged randomly to formula group to develop a new formula and the remainings were used to test the performance of the formula. All participants were instructed to collect a 24-h urine sample, a second morning voiding urine sample (SMU), and a post-meridiem urine sample in the late afternoon or early evening, prior to the evening meal (PMU). All samples were sent to measure sodium and creatinine concentration.2. We compared the differences of office blood pressure, 24-hour ambulatory blood pressure and left ventricular hypertrophy, vascular stiffness and urine protein among groups of different sodium intake. 24hour sodium excretion formulas was obtained using SMU and PMU respectively, which have good cosistency. The difference between the estimated and measured values in sodium excretion is 12.66mmol/day (SMU) and 9.41mmol/day (PM), to be equal to 0.7 g (SMU) and 0.6 g (PM) salt intake. Comparing with Kawasaki and Tanaka method, the new formula shows the lower degree of deviation, and higher accuracy and precision. Blood pressure of high urinary sodium group is higher than that in low urinary sodium group (P < 0.05). Left ventricular hypertrophy and urinary albumin/creatinine aggravated with the salt intake increase, this has eliminated the influence of other factors. All of morphologies of the relationship between ambulatory arterial stiffness index, pulse wave velocity and carotid intima-media thickness with quartiles of sodium intake resembled a J-shaped curve. In Chinese hypertensive population, the

  19. Identification of expanded alleles of the FMR1 gene among high-risk population in Indonesia by using blood spot screening.

    Science.gov (United States)

    Winarni, Tri Indah; Utari, Agustini; Mundhofir, Farmaditya E P; Tong, Tzuhan; Durbin-Johnson, Blythe; Faradz, Sultana M H; Tassone, Flora

    2012-03-01

    The prevalence of Fragile X Syndrome (FXS) is 1 in 4000 in males and 1 in 2500 in males and females, respectively, in the general population. Several screening studies aimed at determining the prevalence of FXS have been conducted in individuals with intellectual disabilities (IDs) with a prevalence varying from 1.15% to 6.3% across different ethnic groups. A previous study in Indonesia showed an FXS prevalence of 1.9% among the ID population. A rapid, effective, and inexpensive method for FMR1 screening, using dried blood spots capable of detecting an expanded FMR1 allele in both males and females, was recently reported. We used this approach to screen 176 blood spots, collected from Central Java, Indonesia, for the presence of expanded FMR1 gene alleles. Samples were collected from high-risk populations: 112 individuals with ID, 32 obtained from individuals with diagnosis of autism spectrum disorders, and 32 individuals with a known family history of FXS. Fourteen subjects carrying an FMR1 expanded allele were identified including 7 premutations (55-200 CGG repeats) and 7 full mutations (>200 repeats). Of the seven subjects identified with a full mutation, one subject was from a non-fragile X family, and six from were families with a history of FXS.

  20. Tay-Sachs and Sandhoff diseases: enzymatic diagnosis in dried blood spots on filter paper: retrospective diagnoses in newborn-screening cards.

    Science.gov (United States)

    Chamoles, Néstor A; Blanco, Mariana; Gaggioli, Daniela; Casentini, Carina

    2002-04-01

    Tay-Sachs disease (TSD), Sandhoff disease (SD) and variants are caused by deficient activity of the lysosomal enzymes hexosaminidase A (HA) and total hexosaminidase (TH) (hexosaminidase A plus B), respectively. For diagnosis, these enzymes are usually measured in plasma or extracts of leukocytes. We describe methods for the assay of hexosaminidase A and total hexosaminidase activities in dried blood spots (DBSs) on filter paper. We studied 163 healthy controls, 9 Tay-Sachs patients, 4 Sandhoff patients, 18 obligate carriers and the newborn-screening cards from two patients with Tay-Sachs and one patient with Sandhoff disease. To tubes containing a 3-mm-diameter blood spot, we added elution liquid and substrate solution. After incubation at 37 degrees C, the amount of hydrolyzed product was compared with a calibrator to allow the quantification of enzyme activity. The described methodology is useful to distinguish patients with Tay-Sachs disease or Sandhoff disease from carriers and controls using samples that are sufficiently stable to be transported to the testing laboratory by mail. The diagnosis of both diseases from a newborn-screening card (NSC) was clearly demonstrated, even after storage for up to 38 months at room temperature. The newborn-screening card has been added to the biological materials that allow the identification of patients with Tay-Sachs disease and Sandhoff disease.

  1. [Diagnostic performance of T-SPOT.TB on peripheral blood in combination with adenosine deaminase on pleural fluid for the diagnosis of tuberculous pleurisy within different age group].

    Science.gov (United States)

    Xu, H Y; Zhang, D Q; Ye, J R; Su, S S; Xie, Y P; Chen, C S; Li, Y P

    2017-06-27

    Objective: To evaluate the performance of T cell enzyme-linked immuno-spot assay (T-SPOT) on peripheral blood in combination with adenosine deaminase (ADA) on pleural fluid for diagnosis of tuberculous (TB) pleurisy within different age groups. Methods: The data of patients with pleural effusion from the Department of Pulmonary and Critical Care Medicine of the First Affiliated Hospital of Wenzhou Medical University from April 2012 to November 2016 were retrospectively analyzed, and the diagnoses of these patients were histopathologically confirmed through medical thoracoscopy. The cases who had confirmed diagnosis, in the same time, received peripheral blood T-SPOT.TB were enrolled. The performance of peripheral blood T-SPOT.TB in combination with pleural fluid ADA on diagnosing TB pleurisy in the younger patients (16-59 years old) and elderly patients (≥60 years old) were analyzed respectively. The sensitivity, specificity and the receiver operating characteristic (ROC) curve were adopted for statistical analysis. Results: A total of 448 cases were finally enrolled, 341(76.1%) confirmed with TB pleurisy, 224 males, 117 females, (47±19) years old; and 107 (23.9%) classified as non-TB pleurisy, 65 males, 42 females, (61±14) years old. There were 285 cases who were classified as younger group, and the other 163 cases were classified as elderly group. The sensitivity and specificity of peripheral blood T-SPOT.TB were 85.4% (204/239) and 71.7% (33/46) in the younger patients, 76.5% (78/102) and 59.0% (36/61) respectively in the elderly patients. The sensitivity of peripheral blood T-SPOT.TB in the younger patients was significantly higher than that in the elderly patients ( P =0.047). The sensitivity and specificity were 99.2% and 95.7% in combination with peripheral blood T-SPOT.TB and pleural fluid ADA respectively in the younger patients. The area under ROC curve (AUC) of T-SPOT.TB in the younger patients was 0.833, AUC of T-SPOT.TB combined with ADA was 0

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

    Science.gov (United States)

    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…

  3. Hematocrit, Anemia, and Arm Preference for Blood Sample ...

    African Journals Online (AJOL)

    and pattern of anemia, as well the arm preferences for blood sample collection among pregnant women in Enugu, South East Nigeria. Subjects and Methods: HCT was determined using venous blood of 200 antenatal women at the University of Nigeria Teaching Hospital (UNTH). Enugu, Nigeria. Questionnaires were used ...

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

    Science.gov (United States)

    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.

  5. Delay in blood sampling for routine newborn screening is associated with increased risk of schizophrenia

    DEFF Research Database (Denmark)

    Nordentoft, Merete; Tidselbak Larsen, Janne; Pedersen, Carsten Bøcker

    2015-01-01

    BACKGROUND: The Danish Neonatal Screening Biobank, containing dried blood spot samples from all newborn in Denmark, is a unique source of data that can be utilized for analyses of genetic and environmental exposures related to schizophrenia and other mental disorders. In previous analyses, we have......, matched on sex and exact date of birth. RESULTS: Both early and late blood sampling was associated with increased risk for schizophrenia. Compared to blood sampling at day 5, sampling at days 0 to 4 after birth was associated with an incidence rate ratio (IRR) of 1.46 (95% CI 1.15-1.87) for development...... admission, and parental education and income, the estimates were slightly different. Thus, blood collection at 0-4days was associated with an IRR of 1.27 (95% CI 0.94-1.71), 6-9days 1.31 (95% CI 0.94-1.84) and 10+days 3.52 (95% CI 1.50 to 8.24). DISCUSSION: After adjusting risk estimates for well-known risk...

  6. BloodSpot: a database of gene expression profiles and transcriptional programs for healthy and malignant haematopoiesis

    DEFF Research Database (Denmark)

    Bagger, Frederik Otzen; Sasivarevic, Damir; Hadi Sohi, Sina

    2016-01-01

    largely inaccessible. Current databases provide information about gene-expression but fail to answer key questions regarding co-regulation, genetic programs or effect on patient survival. To address these shortcomings, we present BloodSpot (www.bloodspot.eu), which includes and greatly extends our...... previously released database HemaExplorer, a database of gene expression profiles from FACS sorted healthy and malignant haematopoietic cells. A revised interactive interface simultaneously provides a plot of gene expression along with a Kaplan–Meier analysis and a hierarchical tree depicting...

  7. Postprandial Insulin and Triglycerides after Different Breakfast Meal Challenges: Use of Finger Stick Capillary Dried Blood Spots to Study Postprandial Dysmetabolism

    Science.gov (United States)

    Kapur, Sonia; Groves, Margaret N.; Zava, David T.; Kapur, Sanjay

    2010-01-01

    Background High levels of insulin and lipids following a meal are recognized risk factors for atherosclerosis. Monitoring such risk factors in the general population is hampered by the inconvenience of venipuncture blood collection, particularly for both premeal and postmeal analyses. This study examined insulin and triglyceride levels in dried blood spots (DBSs) collected after different breakfast meal challenges to assess the potential of this method for risk assessment. Methods Glucose levels were measured using a glucose meter, and insulin and triglycerides were determined in DBS samples collected from 19 healthy volunteers before and at four time points up to 2.5 h after consuming each of five typical breakfast meals varying in nutritional composition. Results At 2 h, glucose was within normal postprandial values (<140 mg/dl) for all meals; significantly lower glucose was seen after meal 2 (the lowest carbohydrate content) compared to the other meals. Insulin returned to normal fasting levels (<15 μIU/ml) in significantly more subjects (90%) after meal 2 and significantly fewer subjects (31%) after meal 4 (highest carbohydrate content) than the other meals. Triglycerides were elevated to a similar extent in all subjects, with no significant differences between meals; levels were still rising at 2.5 h. Conclusions Subjects were able to collect blood spots with minimum disruption to their normal daily activities. Relative ease of collection, analyte stability in dried blood, and the close correlation with serum levels that we have previously demonstrated makes DBS a convenient and simple tool for assessing the individual impact of different diets on postprandial dysmetabolism. PMID:20307382

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

    Science.gov (United States)

    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.

  9. Use of protein: creatinine ratio in a random spot urine sample for predicting significant proteinuria in diabetes mellitus.

    Science.gov (United States)

    Yadav, B K; Adhikari, S; Gyawali, P; Shrestha, R; Poudel, B; Khanal, M

    2010-06-01

    Present study was undertaken during a period of 6 months (September 2008-February 2009) to see an correlation of 24 hours urine protein estimation with random spot protein-creatinine (P:C) ratio among a diabetic patients. The study comprised of 144 patients aged 30-70 years, recruited from Kantipur hospital, Kathmandu. The 24-hr urine sample was collected, followed by spot random urine sample. Both samples were analyzed for protein and creatinine excretion. An informed consent was taken from all participants. Sixteen inadequately collected urine samples as defined by (predicted creatinine--measured creatinine)/predicted creatinine > 0.2 were excluded from analysis. The Spearman's rank correlation between the spot urine P:C ratio and 24-hr total protein were performed by the Statistical Package for Social Service. At the P:C ratio cutoff of 0.15 and reference method (24-hr urine protein) cutoff of 150 mg/day, the correlation coefficient was found to be 0.892 (p urine collection but the cutoff should be carefully selected for different patients group under different laboratory procedures and settings.

  10. Prognostic Significance of Spot Urine Na/K for Longitudinal Changes in Blood Pressure and Renal Function: The Nagahama Study.

    Science.gov (United States)

    Tabara, Yasuharu; Takahashi, Yoshimitsu; Setoh, Kazuya; Kawaguchi, Takahisa; Kosugi, Shinji; Nakayama, Takeo; Matsuda, Fumihiko

    2017-09-01

    Urinary sodium-to-potassium ratio (Na/K) represents a simple measure of sodium load and has been reported to be associated with blood pressure (BP) levels in a cross-sectional setting even with spot measurements. The aim of the present large-scale cohort study is to determine prognostic significance of spot urine Na/K for longitudinal changes in BP levels and renal function. The present study population consisted of 7,063 individuals from the general population. Clinical parameters were measured at baseline and at a follow-up interval of 5 years. Mean systolic BP was slightly increased during the follow-up period (overall, 124 ± 17 to 125 ± 18 mm Hg; nontreated participants, 119 ± 15 to 122 ± 17 mm Hg). Although, the urinary Na/K demonstrated a linear association with BP in a cross-sectional analysis (P K * time interaction, i.e., an intraindividual effect, as an inverse determinant (F = 76.9, P K values at baseline. Spot urine Na/K values were found to be positively associated with renal function in a cross-sectional analysis (P K * time interaction showed inverse associated with renal functional decline (F = 85.8, P K may have limited utility as a prognostic marker of longitudinal BP change, as well as renal functional decline. © American Journal of Hypertension, Ltd 2017. All rights reserved. For Permissions, please email: journals.permissions@oup.com

  11. A prospective clinical trial to compare the performance of dried blood spots prenatal screening for Down's syndrome with conventional non-invasive testing technology.

    Science.gov (United States)

    Hu, Huiying; Jiang, Yulin; Zhang, Minghui; Liu, Shanying; Hao, Na; Zhou, Jing; Liu, Juntao; Zhang, Xiaojin; Ma, Liangkun

    2017-03-01

    To evaluate, side by side, the efficiency of dried blood spots (DBSs) against serum screening for Down's syndrome, and then, to construct a two-tier strategy by topping up the fetal cell-free DNA (cfDNA) secondary screening over the high-risk women marked by the primary blood testing to build a practical screening tactic to identify fetal Down's syndrome. One thousand eight hundred and thirty-seven low-risk Chinese women, with singleton pregnancy, were enrolled for the study. Alpha-fetoprotein and free beta human chorionic gonadotropin were measured for the serum as well as for the parallel DBS samples. Partial high-risk pregnant women identified by primary blood testing (n = 38) were also subject to the secondary cfDNA screening. Diagnostic amniocentesis was utilized to confirm the screening results. The true positive rate for Down's syndrome detection was 100% for both blood screening methods; however, the false-positive rate was 3.0% for DBS and 4.0% for serum screening, respectively. DBS correlated well with serum screening on Down's syndrome detection. Three out of 38 primary high-risk women displayed chromosomal abnormalities by cfDNA analysis, which were confirmed by amniocentesis. Either the true detection rate or the false-positive rate for Down's syndrome between DBS and the serum test is comparable. In addition, blood primary screening aligned with secondary cfDNA analysis, a "before and after" two-tier screening strategy, can massively decrease the false-positive rate, which, then, dramatically reduces the demand for invasive diagnostic operation. Impact statement Children born with Down's syndrome display a wide range of mental and physical disability. Currently, there is no effective treatment to ease the burden and anxiety of the Down's syndrome family and the surrounding society. This study is to evaluate the efficiency of dried blood spots against serum screening for Down's syndrome and to construct a two-tier strategy by topping up the fetal

  12. Comparison of Spot and Time Weighted Averaging (TWA Sampling with SPME-GC/MS Methods for Trihalomethane (THM Analysis

    Directory of Open Access Journals (Sweden)

    Don-Roger Parkinson

    2016-02-01

    Full Text Available Water samples were collected and analyzed for conductivity, pH, temperature and trihalomethanes (THMs during the fall of 2014 at two monitored municipal drinking water source ponds. Both spot (or grab and time weighted average (TWA sampling methods were assessed over the same two day sampling time period. For spot sampling, replicate samples were taken at each site and analyzed within 12 h of sampling by both Headspace (HS- and direct (DI- solid phase microextraction (SPME sampling/extraction methods followed by Gas Chromatography/Mass Spectrometry (GC/MS. For TWA, a two day passive on-site TWA sampling was carried out at the same sampling points in the ponds. All SPME sampling methods undertaken used a 65-µm PDMS/DVB SPME fiber, which was found optimal for THM sampling. Sampling conditions were optimized in the laboratory using calibration standards of chloroform, bromoform, bromodichloromethane, dibromochloromethane, 1,2-dibromoethane and 1,2-dichloroethane, prepared in aqueous solutions from analytical grade samples. Calibration curves for all methods with R2 values ranging from 0.985–0.998 (N = 5 over the quantitation linear range of 3–800 ppb were achieved. The different sampling methods were compared for quantification of the water samples, and results showed that DI- and TWA- sampling methods gave better data and analytical metrics. Addition of 10% wt./vol. of (NH42SO4 salt to the sampling vial was found to aid extraction of THMs by increasing GC peaks areas by about 10%, which resulted in lower detection limits for all techniques studied. However, for on-site TWA analysis of THMs in natural waters, the calibration standard(s ionic strength conditions, must be carefully matched to natural water conditions to properly quantitate THM concentrations. The data obtained from the TWA method may better reflect actual natural water conditions.

  13. An automated blood sampling system used in positron emission tomography

    International Nuclear Information System (INIS)

    Eriksson, L.; Bohm, C.; Kesselberg, M.

    1988-01-01

    Fast dynamic function studies with positron emission tomography (PET), has the potential to give accurate information of physiological functions of the brain. This capability can be realised if the positron camera system accurately quantitates the tracer uptake in the brain with sufficiently high efficiency and in sufficiently short time intervals. However, in addition, the tracer concentration in blood, as a function of time, must be accurately determined. This paper describes and evaluates an automated blood sampling system. Two different detector units are compared. The use of the automated blood sampling system is demonstrated in studies of cerebral blood flow, in studies of the blood-brain barrier transfer of amino acids and of the cerebral oxygen consumption. 5 refs.; 7 figs

  14. Extensive monitoring through multiple blood samples in professional soccer players

    DEFF Research Database (Denmark)

    Heisterberg, Mette F; Fahrenkrug, Jan; Krustrup, Peter

    2013-01-01

    that may be related to changes in training pattern, match exposure or length of the match-season. Especially the end of the preparation-season and at the end of the competitive season seem to be time points were the blood-derived values indicate that the players are under excessive physical strain......ABSTRACT: The aim of this study was to make a comprehensive gathering of consecutive detailed blood samples from professional soccer players, and to analyze different blood parameters in relation to seasonal changes in training and match exposure.Blood samples were collected five times during a six...... months period and analyzed for 37 variables in 27 professional soccer players from the best Danish league. Additionally, players were tested for body composition, VO2max and physical performance by the Yo-Yo intermittent endurance sub-max test (IE2).Multiple variations in blood parameters occurred during...

  15. Delay in blood sampling for routine newborn screening is associated with increased risk of schizophrenia.

    Science.gov (United States)

    Nordentoft, Merete; Larsen, Janne Tidselbak; Pedersen, Carsten Bøcker; Sørensen, Holger Jelling; Hollegaard, Mads Villiam; Hougaard, David Michael; Mortensen, Preben Bo; Petersen, Liselotte

    2015-03-01

    The Danish Neonatal Screening Biobank, containing dried blood spot samples from all newborn in Denmark, is a unique source of data that can be utilized for analyses of genetic and environmental exposures related to schizophrenia and other mental disorders. In previous analyses, we have found that early and late blood sampling, compared to sampling at day 5, was associated with increased risk of schizophrenia. As delay in sampling of blood for neonatal screening cannot in itself influence the risk of schizophrenia, it must be seen as a proxy for unknown underlying causes responsible for this association. Therefore, we investigated whether the increased risk can be explained by other risk factors for schizophrenia. A case-control design was applied. A total of 846 cases with schizophrenia were selected from the Danish Psychiatric Case Register. One control was selected for each case, matched on sex and exact date of birth. Both early and late blood sampling was associated with increased risk for schizophrenia. Compared to blood sampling at day 5, sampling at days 0 to 4 after birth was associated with an incidence rate ratio (IRR) of 1.46 (95% CI 1.15-1.87) for development of schizophrenia, and sampling at days 6 to 9 and at days 10 to 53 was associated with an IRR of 1.5 (95% CI 1.13-1.98) and 3.00 (95% CI 1.59-5.67), respectively. After adjusting the estimates for place of birth, both parents' psychiatric illness, maternal and paternal age, parents' country of origin, child admission, and parental education and income, the estimates were slightly different. Thus, blood collection at 0-4days was associated with an IRR of 1.27 (95% CI 0.94-1.71), 6-9days 1.31 (95% CI 0.94-1.84) and 10+days 3.52 (95% CI 1.50 to 8.24). After adjusting risk estimates for well-known risk factors, delay in sampling of blood for neonatal screening was associated with unexplained increased risk of schizophrenia. Thus, a key finding is that age at test is a proxy for unobserved risk factors

  16. Blood sampling and hemolysis affect concentration of plasma metabolites

    DEFF Research Database (Denmark)

    Theil, Peter Kappel; Pedersen, Lene Juul; Jensen, Margit Bak

    2012-01-01

    design and blood was collected after restraint via vein puncture 1, 4, 11, and 23 h after morning feeding. Plasma samples were categorized as without or with minor or major hemolysis [clear (n = 218), yellow (n = 97), or red (n = 37)] upon centrifugation. Plasma NEFA (P ...Two experiments were carried out to reveal and quantify plasma metabolites that are sensitive to hemolysis and animal stress due to the blood sampling procedure (vein puncture vs. catheter). In Exp. 1, 48 sows were fed 4 diets either once (0800 h) or twice daily (0800 h and 1500 h) in a crossover......, a subset of samples from 24 sows fed twice daily in Exp. 1 was combined with data obtained from 30 sows sampled using jugular vein catheters. All sows in Exp. 2 were fed twice daily (0800 h and 1500 h) and blood samples collected repeatedly 1, 4, 11, and 23 h after morning feeding (other conditions were...

  17. Non-terminal blood sampling techniques in Guinea pigs

    DEFF Research Database (Denmark)

    Birck, Malene Muusfeldt; Tveden-Nyborg, Pernille; Lindblad, Maiken Marie

    2014-01-01

    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 feature...... 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....... 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...

  18. Dried blood spots and parallel artificial liquid membrane extraction-A simple combination of microsampling and microextraction.

    Science.gov (United States)

    Ask, Kristine Skoglund; Øiestad, Elisabeth Leere; Pedersen-Bjergaard, Stig; Gjelstad, Astrid

    2018-06-07

    In this paper, parallel artificial liquid membrane extraction (PALME) was used for the first time to clean-up dried blood spots (DBS) prior to ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Fundamental studies exploring amongst others desorption from the DBS in alkaline or acidic aqueous conditions, total extraction time and absolute recoveries were executed. Desorption and PALME were performed using a set of two 96-well plates, one of them housing the sample and the other comprising the supported liquid membrane (SLM) and the acceptor solution. In one procedure, amitriptyline and quetiapine (basic model analytes) were desorbed from the DBS using 250 μL of 10 mM sodium hydroxide solution (aqueous), and subsequently extracted through the SLM consisting of 4 μL of 1% trioctylamine in dodecyl acetate, and further into an acceptor solution consisting of 50 μL of 20 mM formic acid. In a second procedure, ketoprofen, fenoprofen, flurbiprofen, and ibuprofen (acidic model analytes) were desorbed from the DBS into 20 mM formic acid, extracted through an SLM with dihexyl ether, and further into an acceptor solution of 25 mM ammonia. Within 60 min of PALME, both basic and acidic model analytes were effectively desorbed from the DBS and extracted into the acceptor solution, which was injected directly into the analytical instrument. Recoveries between 63 and 85% for the six model analytes were obtained. PALME provided excellent clean-up from the DBS samples, and acceptor solutions were free from phospholipids. Linearity was obtained with r 2  > 0.99 for five of the six analytes. Accuracy, precision and UHPLC-MS/MS matrix effects were in accordance with the European Medicines Agency (EMA) guideline. Based on these experiments, PALME shows great potential for future processing of DBS in a short and simple way, and with the presented setup, up to 96 DBS can be processed within a total extraction time of 60

  19. Effect of amlodipine, efonidipine, and trichlormethiazide on home blood pressure and upper-normal microalbuminuria assessed by casual spot urine test in essential hypertensive patients.

    Science.gov (United States)

    Hosaka, Miki; Inoue, Ryusuke; Satoh, Michihiro; Watabe, Daisuke; Hanazawa, Tomohiro; Ohkubo, Takayoshi; Asayama, Kei; Obara, Taku; Imai, Yutaka

    2017-11-27

    The aim of this study was to assess the effects of irbesartan alone and combined with amlodipine, efonidipine, or trichlormethiazide on blood pressure (BP) and urinary albumin (UA) excretion in hypertensive patients with microalbuminuria (30≤UA/creatinine (Cr) ratio [UACR] <300 mg/g Cr) and upper-normal microalbuminuria (10≤UACR<30 mg/g Cr). This randomized controlled trial enrolled 175 newly diagnosed and untreated hypertensive patients (home systolic blood pressure [SBP]≥135 mmHg; 10≤UACR<300 mg/g Cr of casual spot urine at the first visit to clinic). All patients were treated with irbesartan (week 0). Patients who failed to achieve home SBP ≤125 mmHg on 8-week irbesartan monotherapy (nonresponders, n = 115) were randomized into three additional drug treatment groups: trichlormethiazide (n = 42), efonidipine (n = 39), or amlodipine (n = 34). Irbesartan monotherapy decreased home SBP and first morning urine samples (morning UACR) for 8 weeks (p < 0.0001). At 8 weeks after randomization, all three additional drugs decreased home SBP (p < 0.0002) and trichlormethiazide significantly decreased morning UACR (p = 0.03). Amlodipine decreased morning UACR in patients with microalbuminuria based on casual spot urine samples (p = 0.048). However, multivariate analysis showed that only higher home SBP and UACR at week 8, but not any additional treatments, were significantly associated with UACR reduction between week 8 and week 16. In conclusion, crucial points of the effects of combination therapy on UACR were basal UACR and SBP levels. The effect of trichlormethiazide or amlodipine treatment in combination with irbesartan treatment on microalbuminuria needs to be reexamined based on a larger sample size after considering basal UACR and SBP levels.

  20. Validation and modification of dried blood spot-based glycosylated hemoglobin assay for the longitudinal aging study in India.

    Science.gov (United States)

    Hu, Peifeng; Edenfield, Michael; Potter, Alan; Kale, Varsha; Risbud, Arun; Williams, Sharon; Lee, Jinkook; Bloom, David E; Crimmins, Eileen; Seeman, Teresa

    2015-01-01

    This study aims to validate a modified dried blood spot (DBS)-based glycosylated hemoglobin (HbA1c) assay protocol, after a pretest in India showed poor correlation between the original DBS-based protocol and venous results. The original protocol was tested on different chemistry analyzers and then simplified at the University of Washington (UW). A second pretest was conducted in India to validate the modified assay protocol, using 44 quality control specimens. Data from UW indicated that, using the original protocol, the correlation coefficients between DBS and venous results were above 0.98 on both Bio-Rad and Olympus chemistry analyzers. The protocol worked equally well on filter paper, with or without pre-treatment, and when the recommended amount of blood spot material, or less, was used. A second pretest of the modified protocol confirmed that DBS-based levels from both Olympus and Roche chemistry analyzers were well correlated with DBS results from UW (correlation coefficients were above 0.96), as well as with venous values (correlation coefficients were above 0.94). The DBS-based HbA1c values are highly correlated with venous results. The pre-treatment of filter paper does not appear to be necessary. The poor results from the first pretest are probably due to factors unrelated to the protocol, such as problems with the chemistry analyzer or assay reagents. © 2015 Wiley Periodicals, Inc.

  1. Prevalence of Congenital Cytomegalovirus Infection Assessed Through Viral Genome Detection in Dried Blood Spots in Children with Autism Spectrum Disorders.

    Science.gov (United States)

    Gentile, Ivan; Zappulo, Emanuela; Riccio, Maria Pia; Binda, Sandro; Bubba, Laura; Pellegrinelli, Laura; Scognamiglio, Domenico; Operto, Francesca; Margari, Lucia; Borgia, Guglielmo; Bravaccio, Carmela

    2017-01-01

    Autism spectrum disorders (ASD) are neurodevelopmental disorders without a definitive etiology in most cases. Environmental factors, such as viral infections, have been linked with anomalies in brain growth, neuronal development, and functional connectivity. Congenital cytomegalovirus (CMV) infection has been associated with the onset of ASD in several case reports. The aim of this study was to evaluate the prevalence of congenital CMV infection in children with ASD and in healthy controls. The CMV genome was tested by polymerase chain reaction (PCR) on dried blood spots collected at birth from 82 children (38 with ASD and 44 controls). The prevalence of congenital CMV infection was 5.3% (2/38) in cases and 0% (0/44) in controls (p=0.212). The infection rate was about 10-fold higher in patients with ASD than in the general Italian population at birth. For this reason, detection of CMV-DNA on dried blood spots could be considered in the work-up that is usually performed at ASD diagnosis to rule-out a secondary form. Given the potential prevention and treatment of CMV infection, this study could have intriguing consequences, at least for a group of patients with ASD. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

  2. Detection of antileishmanial antibodies in blood sampled from blood bank donors in Istanbul.

    Science.gov (United States)

    Ates, Sezen Canim; Bagirova, Malahat; Allahverdiyev, Adil M; Baydar, Serap Yesilkir; Koc, Rabia Cakir; Elcicek, Serhat; Abamor, Emrah Sefik; Oztel, Olga Nehir

    2012-06-01

    According to the WHO, only 5-20% of the total cases of leishmaniasis are symptomatic leishmaniasis; the other cases are identified as asymptomatic leishmaniasis. In recent studies, it has been demonstrated that donor blood plays an important role in the epidemiology of asymptomatic leishmaniasis. However, the number of the studies on this subject is still insufficient. Additionally, donor blood samples obtained from Istanbul, which is the biggest metropolitan area in Turkey, have not been investigated with regard to Leishmania. Moreover, there is no information about the sensitivity of noninvasive serological methods that are used in the detection of leishmaniasis donor blood samples. Accordingly, this study aimed to investigate the presence of antileishmanial antibodies in blood samples obtained from blood bank donors in Istanbul, by using different serologic methods, and to determine the most sensitive detection method. Blood samples were taken from 188 healthy blood bank donors to the Capa Turkish Red Crescent Blood Bank (Istanbul, Turkey), and the presence of antileishmanial antibodies was measured by indirect immunofluorescent antibody test (IFAT), ELISA, immunochromatographic dipstick rapid test, and western blot (WB). Antileishmanial antibodies were determined in 12 out of 188 samples by IFAT (6.4%), and six out of these 12 donors were found to be positive at diagnostic titer 1:128 (3.2%). One hundred and eighty eight samples were investigated by ELISA and one (0.5%) of them gave a positive result. None of 188 samples provided a positive result by immunochromatographic test. WB applied to the 12 seroreactive donors showed that three out of 12 donors were positive. In this study, the presence of antileishmanial antibodies in blood samples of blood bank donors from Istanbul has been demonstrated by using feasible and low-cost serological methods. Additionally, in comparison with other simple and low-cost detection methods, WB was used for confirmation. IFAT

  3. The use of dried blood spots for quantification of 15 antipsychotics and 7 metabolites with ultra-high performance liquid chromatography - tandem mass spectrometry.

    Science.gov (United States)

    Patteet, Lisbeth; Maudens, Kristof E; Stove, Christophe P; Lambert, Willy E; Morrens, Manuel; Sabbe, Bernard; Neels, Hugo

    2015-06-01

    Therapeutic drug monitoring of antipsychotics is important in optimizing individual therapy. In psychiatric populations, classical venous blood sampling is experienced as frightening. Interest in alternative techniques, like dried blood spots (DBS), has consequently increased. A fast and easy to perform DBS method for quantification of 16 antipsychotics (amisulpride, aripiprazole, asenapine, bromperidol, clozapine, haloperidol, iloperidone, levosulpiride, lurasidone, olanzapine, paliperidone, pipamperone, quetiapine, risperidone, sertindole and zuclopenthixol) and 8 metabolites was developed. DBS were prepared using 25 μL of whole blood and extraction of complete spots was performed using methanol: methyl-t-butyl-ether (4:1). After evaporation, the extract was reconstituted in the mobile phase and 10 μL were injected on an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Separation using a C18 column and gradient elution with a flow rate of 0.5 mL/min resulted in a 6-min run-time. Ionization was performed in positive mode and a dynamic MRM method was applied. Median recovery was 66.4 % (range 28.7-84.5%). Accuracy was within the acceptance criteria, except for pipamperone (LLOQ and low concentration) and lurasidone (low concentration). Imprecision was only aberrant for lurasidone at low and medium concentration. All compounds were stable during 1 month at room temperature, 4 °C and -18 °C. Lurasidone was unstable when the extract was stored for 12 h on the autosampler. Absolute matrix effects (ME) (median 66.1%) were compensated by the use of deuterated IS (median 98.8%). The DBS method was successfully applied on 25-μL capillary DBS from patients and proved to be a reliable alternative for quantification of all antipsychotics except for olanzapine and N-desmethylolanzapine. Copyright © 2014 John Wiley & Sons, Ltd.

  4. Validation and Assessment of Three Methods to Estimate 24-h Urinary Sodium Excretion from Spot Urine Samples in High-Risk Elder Patients of Stroke from the Rural Areas of Shaanxi Province.

    Science.gov (United States)

    Ma, Wenxia; Yin, Xuejun; Zhang, Ruijuan; Liu, Furong; Yang, Danrong; Fan, Yameng; Rong, Jie; Tian, Maoyi; Yu, Yan

    2017-10-11

    Background : 24-h urine collection is regarded as the "gold standard" for monitoring sodium intake at the population level, but ensuring high quality urine samples is difficult to achieve. The Kawasaki, International Study of Sodium, Potassium, and Blood Pressure (INTERSALT) and Tanaka methods have been used to estimate 24-h urinary sodium excretion from spot urine samples in some countries, but few studies have been performed to compare and validate these methods in the Chinese population. Objective : To compare and validate the Kawasaki, INTERSALT and Tanaka formulas in predicting 24-h urinary sodium excretion using spot urine samples in 365 high-risk elder patients of strokefrom the rural areas of Shaanxi province. Methods : Data were collected from a sub-sample of theSalt Substitute and Stroke Study. 365 high-risk elder patients of stroke from the rural areas of Shaanxi province participated and their spot and 24-h urine specimens were collected. The concentrations of sodium, potassium and creatinine in spot and 24-h urine samples wereanalysed. Estimated 24-h sodium excretion was predicted from spot urine concentration using the Kawasaki, INTERSALT, and Tanaka formulas. Pearson correlation coefficients and agreement by Bland-Altman method were computed for estimated and measured 24-h urinary sodium excretion. Results : The average 24-h urinary sodium excretion was 162.0 mmol/day, which representing a salt intake of 9.5 g/day. Three predictive equations had low correlation with the measured 24-h sodium excretion (r = 0.38, p h sodium excretion were observed (all p h sodium excretion. Conclusion : The Kawasaki, INTERSALT and Tanaka methods for estimation of 24-h urinary sodium excretion from spot urine specimens were inadequate for the assessment of sodium intake at the population level in high-risk elder patients of stroke from the rural areas of Shaanxi province, although the Kawasaki method was the least biased compared with the other two methods.

  5. Extensive monitoring through multiple blood samples in professional soccer players.

    Science.gov (United States)

    Heisterberg, Mette F; Fahrenkrug, Jan; Krustrup, Peter; Storskov, Anders; Kjær, Michael; Andersen, Jesper L

    2013-05-01

    The aim of this study was to make a comprehensive gathering of consecutive detailed blood samples from professional soccer players and to analyze different blood parameters in relation to seasonal changes in training and match exposure. Blood samples were collected 5 times during a 6-month period and analyzed for 37 variables in 27 professional soccer players from the best Danish league. Additionally, the players were tested for body composition, V[Combining Dot Above]O2max and physical performance by the Yo-Yo intermittent endurance submax test (IE2). Multiple variations in blood parameters occurred during the observation period, including a decrease in hemoglobin and an increase in hematocrit as the competitive season progressed. Iron and transferrin were stable, whereas ferritin showed a decrease at the end of the season. The immunoglobulin A (IgA) and IgM increased in the period with basal physical training and at the end of the season. Leucocytes decreased with increased physical training. Lymphocytes decreased at the end of the season. The V[Combining Dot Above]O2max decreased toward the end of the season, whereas no significant changes were observed in the IE2 test. The regular blood samples from elite soccer players reveal significant changes that may be related to changes in training pattern, match exposure, or length of the match season. Especially the end of the preparation season and at the end of the competitive season seem to be time points were the blood-derived values indicate that the players are under excessive physical strain and might be more subjected to a possible overreaching-overtraining conditions. We suggest that regular analyses of blood samples could be an important initiative to optimize training adaptation, training load, and game participation, but sampling has to be regular, and a database has to be built for each individual player.

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

    Science.gov (United States)

    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.

  7. Neonatal detection of Aicardi Goutières Syndrome by increased C26:0 lysophosphatidylcholine and interferon signature on newborn screening blood spots.

    Science.gov (United States)

    Armangue, Thais; Orsini, Joseph J; Takanohashi, Asako; Gavazzi, Francesco; Conant, Alex; Ulrick, Nicole; Morrissey, Mark A; Nahhas, Norah; Helman, Guy; Gordish-Dressman, Heather; Orcesi, Simona; Tonduti, Davide; Stutterd, Chloe; van Haren, Keith; Toro, Camilo; Iglesias, Alejandro D; van der Knaap, Marjo S; Goldbach Mansky, Raphaela; Moser, Anne B; Jones, Richard O; Vanderver, Adeline

    2017-11-01

    Aicardi Goutières Syndrome (AGS) is a heritable interferonopathy associated with systemic autoinflammation causing interferon (IFN) elevation, central nervous system calcifications, leukodystrophy and severe neurologic sequelae. An infant with TREX1 mutations was recently found to have abnormal C26:0 lysophosphatidylcholine (C26:0 Lyso-PC) in a newborn screening platform for X-linked adrenoleukodystrophy, prompting analysis of this analyte in retrospectively collected samples from individuals affected by AGS. In this study, we explored C26:0 Lyso-PC levels and IFN signatures in newborn blood spots and post-natal blood samples in 19 children with a molecular and clinical diagnosis of AGS and in the blood spots of 22 healthy newborns. We used Nanostring nCounter™ for IFN-induced gene analysis and a high-performance liquid chromatography with tandem mass spectrometry (HPLC MS/MS) newborn screening platform for C26:0 Lyso-PC analysis. Newborn screening cards from patients across six AGS associated genes were collected, with a median disease presentation of 2months. Thirteen out of 19 (68%) children with AGS had elevations of first tier C26:0 Lyso-PC (>0.4μM), that would have resulted in a second screen being performed in a two tier screening system for X-linked adrenoleukodystrophy (X-ALD). The median (95%CI) of first tier C26:0 Lyso-PC values in AGS individuals (0.43μM [0.37-0.48]) was higher than that seen in controls (0.21μM [0.21-0.21]), but lower than X-ALD individuals (0.72μM [0.59-0.84])(p<0.001). Fourteen of 19 children had elevated expression of IFN signaling on blood cards relative to controls (Sensitivity 73.7%, 95%CI 51-88%, Specificity 95%, 95% CI 78-99%) including an individual with delayed disease presentation (36months of age). All five AGS patients with negative IFN signature at birth had RNASEH2B mutations. Consistency of agreement between IFN signature in neonatal and post-natal samples was high (0.85). This suggests that inflammatory markers

  8. Does volumetric absorptive microsampling eliminate the hematocrit bias for caffeine and paraxanthine in dried blood samples? A comparative study.

    Science.gov (United States)

    De Kesel, Pieter M M; Lambert, Willy E; Stove, Christophe P

    2015-06-30

    Volumetric absorptive microsampling (VAMS) is a novel sampling technique that allows the straightforward collection of an accurate volume of blood (approximately 10μL) from a drop or pool of blood by dipping an absorbent polymeric tip into it. The resulting blood microsample is dried and analyzed as a whole. The aim of this study was to evaluate the potential of VAMS to overcome the hematocrit bias, an important issue in the analysis of dried blood microsamples. An LC-MS/MS method for analysis of the model compounds caffeine and paraxanthine in VAMS samples was fully validated and fulfilled all pre-established criteria. In conjunction with previously validated procedures for dried blood spots (DBS) and blood, this allowed us to set up a meticulous comparative study in which both compounds were determined in over 80 corresponding VAMS, DBS and liquid whole blood samples. These originated from authentic human patient samples, covering a wide hematocrit range (0.21-0.50). By calculating the differences with reference whole blood concentrations, we found that analyte concentrations in VAMS samples were not affected by a bias that changed over the evaluated hematocrit range, in contrast to DBS results. However, VAMS concentrations tend to overestimate whole blood concentrations, as a consistent positive bias was observed. A different behavior of VAMS samples prepared from incurred and spiked blood, combined with a somewhat reduced recovery of caffeine and paraxanthine from VAMS tips at high hematocrit values, an effect that was not observed for DBS using a very similar extraction procedure, was found to be at the basis of the observed VAMS-whole blood deviations. Based on this study, being the first in which the validity and robustness of VAMS is evaluated by analyzing incurred human samples, it can be concluded that VAMS effectively assists in eliminating the effect of hematocrit. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  9. Single versus duplicate blood samples in ACTH stimulated adrenal vein sampling

    NARCIS (Netherlands)

    Dekkers, T.; Arntz, M.; Wilt, G.J. van der; Schultze Kool, L.J.; Sweep, F.C.; Hermus, A.R.M.M.; Lenders, J.W.M.; Deinum, J.

    2013-01-01

    BACKGROUND: Adrenal vein sampling (AVS) is the preferred test for subtyping primary aldosteronism. However, the procedure is technically demanding and costly. In AVS it is common practice to take duplicate blood samples at each location. In this paper we explore whether a single sample procedure

  10. Correction for the Hematocrit Bias in Dried Blood Spot Analysis Using a Nondestructive, Single-Wavelength Reflectance-Based Hematocrit Prediction Method.

    Science.gov (United States)

    Capiau, Sara; Wilk, Leah S; De Kesel, Pieter M M; Aalders, Maurice C G; Stove, Christophe P

    2018-02-06

    The hematocrit (Hct) effect is one of the most important hurdles currently preventing more widespread implementation of quantitative dried blood spot (DBS) analysis in a routine context. Indeed, the Hct may affect both the accuracy of DBS methods as well as the interpretation of DBS-based results. We previously developed a method to determine the Hct of a DBS based on its hemoglobin content using noncontact diffuse reflectance spectroscopy. Despite the ease with which the analysis can be performed (i.e., mere scanning of the DBS) and the good results that were obtained, the method did require a complicated algorithm to derive the total hemoglobin content from the DBS's reflectance spectrum. As the total hemoglobin was calculated as the sum of oxyhemoglobin, methemoglobin, and hemichrome, the three main hemoglobin derivatives formed in DBS upon aging, the reflectance spectrum needed to be unmixed to determine the quantity of each of these derivatives. We now simplified the method by only using the reflectance at a single wavelength, located at a quasi-isosbestic point in the reflectance curve. At this wavelength, assuming 1-to-1 stoichiometry of the aging reaction, the reflectance is insensitive to the hemoglobin degradation and only scales with the total amount of hemoglobin and, hence, the Hct. This simplified method was successfully validated. At each quality control level as well as at the limits of quantitation (i.e., 0.20 and 0.67) bias, intra- and interday imprecision were within 10%. Method reproducibility was excellent based on incurred sample reanalysis and surpassed the reproducibility of the original method. Furthermore, the influence of the volume spotted, the measurement location within the spot, as well as storage time and temperature were evaluated, showing no relevant impact of these parameters. Application to 233 patient samples revealed a good correlation between the Hct determined on whole blood and the predicted Hct determined on venous DBS. The

  11. Fetal scalp blood sampling in labor - a review

    DEFF Research Database (Denmark)

    Jørgensen, Jan Stener; Weber, Tom

    2014-01-01

    During the 1970s and 1980s, electronic fetal monitoring and fetal scalp blood sampling (FBS) were introduced without robust evidence. With a methodical review of the published literature, and using one randomized controlled trial, seven controlled studies, nine randomized studies of various...... in the interpretation of CTG patterns. This article is protected by copyright. All rights reserved....

  12. A simple method for repetitive blood sampling of cattle

    African Journals Online (AJOL)

    A simple practical technique is described for the collection of blood samples from at least six catile concurrenily at short time interuals for up to 48 h. The technique is advantageous in that it is carried out under minimum srress conditions. 'n Eenvoudige praktiese tegniek vir die gelyktydige trekking van bloedmonsters van ten ...

  13. Efficiency of Extraction of Trace metals from Blood samples using ...

    African Journals Online (AJOL)

    MICHAEL HORSFALL

    Efficiency of Extraction of Trace metals from Blood samples using Wet Digestion and. Microwave Digestion Techniques. *1M. I. YAHAYA; A. SHEHU; F.G. DABAI. Department of Applied Chemistry, Federal University Dutsin – Ma, Katsina State, Nigeria. Biology Department, Kebbi State College of Basic and Advanced Studies ...

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

    Science.gov (United States)

    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.

  15. A comparison of DNA extraction protocols from blood spotted on FTA cards for the detection of tick-borne pathogens by Reverse Line Blot hybridization.

    Science.gov (United States)

    Hailemariam, Zerihun; Ahmed, Jabbar Sabir; Clausen, Peter-Henning; Nijhof, Ard Menzo

    2017-01-01

    An essential step in the molecular detection of tick-borne pathogens (TBPs) in blood is the extraction of DNA. When cooled storage of blood under field conditions prior to DNA extraction in a dedicated laboratory is not possible, the storage of blood on filter paper forms a promising alternative. We evaluated six DNA extraction methods from blood spotted on FTA Classic ® cards (FTA cards), to determine the optimal protocol for the subsequent molecular detection of TBPs by PCR and the Reverse Line Blot hybridization assay (RLB). Ten-fold serial dilutions of bovine blood infected with Babesia bovis, Theileria mutans, Anaplasma marginale or Ehrlichia ruminantium were made by dilution with uninfected blood and spotted on FTA cards. Subsequently, DNA was extracted from FTA cards using six different DNA extraction protocols. DNA was also isolated from whole blood dilutions using a commercial kit. PCR/RLB results showed that washing of 3mm discs punched from FTA cards with FTA purification reagent followed by DNA extraction using Chelex ® resin was the most sensitive procedure. The detection limit could be improved when more discs were used as starting material for the DNA extraction, whereby the use of sixteen 3mm discs proved to be most practical. The presented best practice method for the extraction of DNA from blood spotted on FTA cards will facilitate epidemiological studies on TBPs. It may be particularly useful for field studies where a cold chain is absent. Copyright © 2016 Elsevier GmbH. All rights reserved.

  16. Urinary iodine concentration follows a circadian rhythm: a study with 3023 spot urine samples in adults and children.

    Science.gov (United States)

    Als, C; Helbling, A; Peter, K; Haldimann, M; Zimmerli, B; Gerber, H

    2000-04-01

    Our overall aim is to monitor iodine supply in a prospective study before and after the September 1998 increase of salt iodide content in Switzerland. Because iodide is supplied by alimentation, we moreover wondered whether urinary iodine concentration (UI) is governed by circadian rhythmicity. Forty-two subjects (18 males and 24 females, including 13 children) collected 3023 urine spots between May 1996 and May 1998, at a rate of three to five samples per month, at any time of the day. The results show that circadian rhythmicity of UI in adults and children was found independent of the individual subject, age, gender, and season. Lowest UI levels were found between 8-11 h. A curve increasing progressively between 12 and 24 h was obtained. UI returned to base-line levels between 21 and 22 h in children only. UI peaks occurred 4-5 h after main meals; children's peaks occurred later than that of adults. Although the existence of a circadian rhythm of UI is probably universal, its profile, however, depends on alimentation. Because nadir of UI is represented by morning spots, this might seem an appropriate collecting period. In view of the significant circadian rhythmicity of UI, studies with restriction of sampling time to morning hours, for example, cannot be directly compared with studies in which urine is sampled all over the day.

  17. Whole blood solubilization and discoloration before LSC of yttrium samples

    International Nuclear Information System (INIS)

    Lima, Marina F.; Leonardo, Lucio

    2009-01-01

    Liquid scintillation counting of whole blood and animal tissues samples could be severely impaired owing to quenching by their compounds. The objective of this previous study is preparing one protocol of 90 Y measurement to apply in biodistribution and dosimetry studies of radiopharmaceuticals labeled with this isotope and other beta emitters in in vivo and ex vivo samples. The first parameters considered to choose a method were: the largest blood sample per collection (80μl-90μl), attending the collection limit of less than 7.5% of total circulating blood volume for in vivo samples. Other parameters were the use of EDTA and cyclohexydine as solubilization and lytic agents, HNO 3 and H 2 O 2 as mineralizing agents and NH 4 OH as neutralization agent. One samples batch was tested in a water bath under the lower temperature to prevent the volume lose of the ionic phase. Other samples batch was mineralized over a hot-plate at 120 deg C following the currently largest sample amounts processing procedure in our laboratory by using HNO 3 and H 2 O 2 . Results show the contribution of the blood fragments as quenching in the region A ( 3 and/or NH 4 OH in the hot-plate digestion. As expected, the measurements in the three spectral regions show the proteins and colored fragments were completely removed by the hot-plate digestion. The rate between efficiency and 90 Sr- 90 Y concentration had not significant differences in the range between 120 Bq and 1200 Bq. (author)

  18. Evaluation of portable blood glucose meters using canine and feline pooled blood samples.

    Science.gov (United States)

    Mori, A; Oda, H; Onozawa, E; Shono, S; Takahashi, T; Yamashita, S; Fujimoto, H; Sako, T

    2016-12-01

    This study evaluated the accuracy and reproducibility of a human portable blood glucose meter (PBGM) for canine and feline whole blood. Reference plasma glucose values (RPGV) were concurrently measured using glucose oxidation methods. Fifteen healthy dogs and 6 healthy cats were used for blood sampling. Blood glucose concentrations and hematocrits were adjusted using pooled blood samples for our targeted values. A positive correlation between the PBGM and RPGV was found for both dogs (y = 0.877, x = -24.38, r = 0.9982, n = 73) and cats (y = 1.048, x = -27.06, r = 0.9984, n = 69). Acceptable results were obtained in error grid analysis between PBGM and RPGV in both dogs and cats; 100% of these results were within zones A and B. Following ISO recommendations, a PBGM is considered accurate if 95% of the measurements are within ± 15 mg/dl of the RPGV when the glucose concentration is cats (39%, 27 of 69 cats). Blood samples with high hematocrits induced lower whole blood glucose values measured by the PBGM than RPGV under hypoglycemic, normoglycemic, and hyperglycemic conditions in both dogs and cats. Therefore, this device is not clinically useful in dogs and cats. New PBGMs which automatically compensate for the hematocrit should be developed in veterinary practice.

  19. 3-Methylcrotonyl-coenzyme A carboxylase deficiency in Amish/Mennonite adults identified by detection of increased acylcarnitines in blood spots of their children.

    Science.gov (United States)

    Gibson, K M; Bennett, M J; Naylor, E W; Morton, D H

    1998-03-01

    Isolated 3-methylcrotonyl coenzyme A carboxylase (MCC) deficiency was documented in four adult women from the Amish/Mennonite population of Lancaster County, Pennsylvania. Metabolic and enzymatic investigations in these individuals were instituted after the detection of abnormal acylcarnitine profiles in blood spots obtained from their newborn children, in whom MCC activity was normal.

  20. An LC-MS/MS-Based Method for the Quantification of Pyridox(am)ine 5'-Phosphate Oxidase Activity in Dried Blood Spots from Patients with Epilepsy.

    Science.gov (United States)

    Wilson, Matthew P; Footitt, Emma J; Papandreou, Apostolos; Uudelepp, Mari-Liis; Pressler, Ronit; Stevenson, Danielle C; Gabriel, Camila; McSweeney, Mel; Baggot, Matthew; Burke, Derek; Stödberg, Tommy; Riney, Kate; Schiff, Manuel; Heales, Simon J R; Mills, Kevin A; Gissen, Paul; Clayton, Peter T; Mills, Philippa B

    2017-09-05

    We report the development of a rapid, simple, and robust LC-MS/MS-based enzyme assay using dried blood spots (DBS) for the diagnosis of pyridox(am)ine 5'-phosphate oxidase (PNPO) deficiency (OMIM 610090). PNPO deficiency leads to potentially fatal early infantile epileptic encephalopathy, severe developmental delay, and other features of neurological dysfunction. However, upon prompt treatment with high doses of vitamin B 6 , affected patients can have a normal developmental outcome. Prognosis of these patients is therefore reliant upon a rapid diagnosis. PNPO activity was quantified by measuring pyridoxal 5'-phosphate (PLP) concentrations in a DBS before and after a 30 min incubation with pyridoxine 5'-phosphate (PNP). Samples from 18 PNPO deficient patients (1 day-25 years), 13 children with other seizure disorders receiving B 6 supplementation (1 month-16 years), and 37 child hospital controls (5 days-15 years) were analyzed. DBS from the PNPO-deficient samples showed enzyme activity levels lower than all samples from these two other groups as well as seven adult controls; no false positives or negatives were identified. The method was fully validated and is suitable for translation into the clinical diagnostic arena.

  1. High efficacy of first-line ART in a West African cohort, assessed by dried blood spot virological and pharmacological measurements.

    Science.gov (United States)

    de Truchis, Pierre; Lê, Minh Patrick; Daou, Mamane; Madougou, Boubacar; Nouhou, Yacouba; Moussa Saley, Sahada; Sani, Achirou; Adehossi, Eric; Rouveix, Elisabeth; Saidou, Mamadou; Peytavin, Gilles; Delaugerre, Constance

    2016-11-01

    The objectives of this study were to determine the rate of viral success in HIV-infected patients on first-line ART by the assessment of dried blood spot (DBS) viral load (VL) and to assess the performance of DBS sampling for VL measurement, genotypic resistance and antiretroviral concentration determinations. HIV-infected patients treated for >1 year with first-line ART in Niamey, Niger were included. VL based on nucleic acid sequence-based amplification (NASBA) assay (limit of quantification 100 copies/mL); antiretroviral concentrations were interpreted using standard plasma cut-offs after extrapolation of blood to plasma results. Median (IQR) results are presented. Two hundred and eighteen patients (61% women), aged 41 (34-46) years, with 138 (56-235) CD4 cells/mm 3 at baseline were included. After 4 (2-6) years of follow-up under therapy, CD4 gain was +197 (98-372) cells/mm 3 ; 81% had VL ART in Niger. Adherence was high, according to antiretroviral concentrations, and the majority of failures were explained by selection of drug resistance mutations detected in the DBS genotype. Using DBS might improve the assessment of ART failure in HIV-infected patients in low-income countries. © The Author 2016. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  2. Genome-wide DNA methylation profiling with MeDIP-seq using archived dried blood spots

    DEFF Research Database (Denmark)

    Staunstrup, Nicklas H; Starnawska, Anna; Nyegaard, Mette

    2016-01-01

    biobanks. However, availability of this biological material is highly limited as each DBS is made only from a few droplets of blood and storage conditions may be suboptimal for epigenetic studies. Furthermore, as relevant markers may reside outside gene bodies, epigenome-wide interrogation is needed....... RESULTS: Here we demonstrate, as a proof of principle, that genome-wide interrogation of the methylome based on methylated DNA immunoprecipitation coupled with next-generation sequencing (MeDIP-seq) is feasible using a single 3.2 mm DBS punch (60 ng DNA) from filter cards archived for up to 16 years...

  3. Genetic screening of spinal muscular atrophy using a real-time modified COP-PCR technique with dried blood-spot DNA.

    Science.gov (United States)

    Ar Rochmah, Mawaddah; Harahap, Nur Imma Fatimah; Niba, Emma Tabe Eko; Nakanishi, Kenta; Awano, Hiroyuki; Morioka, Ichiro; Iijima, Kazumoto; Saito, Toshio; Saito, Kayoko; Lai, Poh San; Takeshima, Yasuhiro; Takeuchi, Atsuko; Bouike, Yoshihiro; Okamoto, Maya; Nishio, Hisahide; Shinohara, Masakazu

    2017-10-01

    Spinal muscular atrophy (SMA) is a common neuromuscular disorder caused by mutations in SMN1. More than 95% of SMA patients carry homozygous SMN1 deletion. SMA is the leading genetic cause of infant death, and has been considered an incurable disease. However, a recent clinical trial with an antisense oligonucleotide drug has shown encouraging clinical efficacy. Thus, early and accurate detection of SMN1 deletion may improve prognosis of many infantile SMA patients. A total of 88 DNA samples (37 SMA patients, 12 carriers and 39 controls) from dried blood spots (DBS) on filter paper were analyzed. All participants had previously been screened for SMN genes by PCR restriction fragment length polymorphism (PCR-RFLP) using DNA extracted from freshly collected blood. DNA was extracted from DBS that had been stored at room temperature (20-25°C) for 1week to 5years. To ensure sufficient quality and quantity of DNA samples, target sequences were pre-amplified by conventional PCR. Real-time modified competitive oligonucleotide priming-PCR (mCOP-PCR) with the pre-amplified PCR products was performed for the gene-specific amplification of SMN1 and SMN2 exon 7. Compared with PCR-RFLP using DNA from freshly collected blood, results from real-time mCOP-PCR using DBS-DNA for detection of SMN1 exon 7 deletion showed a sensitivity of 1.00 (CI [0.87, 1.00])] and specificity of 1.00 (CI [0.90, 1.00]), respectively. We combined DNA extraction from DBS on filter paper, pre-amplification of target DNA, and real-time mCOP-PCR to specifically detect SMN1 and SMN2 genes, thereby establishing a rapid, accurate, and high-throughput system for detecting SMN1-deletion with practical applications for newborn screening. Copyright © 2017 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.

  4. High-Quality Exome Sequencing of Whole-Genome Amplified Neonatal Dried Blood Spot DNA

    DEFF Research Database (Denmark)

    Poulsen, Jesper Buchhave; Lescai, Francesco; Grove, Jakob

    2016-01-01

    be amplified to obtain micrograms of an otherwise limited resource, referred to as whole-genome amplified DNA (wgaDNA). Here we investigate the robustness of exome sequencing of wgaDNA of neonatal DBS samples. We conducted three pilot studies of seven, eight and seven subjects, respectively. For each subject...... from variant calls. No differences were observed substituting 2x3.2 with 2x1.6 mm discs, allowing for additional reduction of sample material in future projects....

  5. Newborn blood spot screening for sickle cell disease by using tandem mass spectrometry: implementation of a protocol to identify only the disease states of sickle cell disease.

    Science.gov (United States)

    Moat, Stuart J; Rees, Derek; King, Lawrence; Ifederu, Adeboye; Harvey, Katie; Hall, Kate; Lloyd, Geoff; Morrell, Christine; Hillier, Sharon

    2014-02-01

    The currently recommended technologies of HPLC and isoelectric focusing for newborn blood spot screening for sickle cell disease (SCD) identify both the disease and carrier states, resulting in large numbers of infants being followed up unnecessarily. Analysis of blood spot tryptic peptides performed by using tandem mass spectrometry (MS/MS) is an alternative technology to detect hemoglobin (Hb) variant disorders. We analyzed 2154 residual newborn blood spots and 675 newborn blood spots from infants with Hb variants by using MS/MS after trypsin digestion. Screening cutoffs were developed by using the ratio between the variant peptide-to-wild-type peptide abundance for HbS, C, D(Punjab), O(Arab), Lepore, and E peptides. A postanalytical data analysis protocol was developed using these cutoffs to detect only the disease states of SCD and not to identify carrier states. A parallel study of 13 249 newborn blood spots from a high-prevalence SCD area were analyzed by both MS/MS and HPLC. Screening cutoffs developed distinguished the infants with the disease states of SCD, infants who were carriers of SCD, and infants with normal Hb. In the parallel study no false-negative results were identified, and all clinically relevant cases were correctly identified using the MS/MS protocol. Unblinding the data revealed a total of 328 carrier infants that were successfully excluded by the protocol. The screening protocol developed correctly identified infants with the disease states of SCD. Furthermore, large numbers of sickle cell carrier infants were successfully not identified, thereby avoiding unnecessary follow-up testing and referral for genetic counseling.

  6. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric imaging of synthetic polymer sample spots prepared using ionic liquid matrices.

    Science.gov (United States)

    Gabriel, Stefan J; Pfeifer, Dietmar; Schwarzinger, Clemens; Panne, Ulrich; Weidner, Steffen M

    2014-03-15

    Polymer sample spots for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) prepared by the dried-droplet method often reveal ring formation accompanied by possible segregation of matrix and sample molecules as well as of the polymer homologs itself. Since the majority of sample spots are prepared by this simple and fast method, a matrix or sample preparation method that excludes such segregation has to be found. Three different ionic liquid matrices based on conventionally used aromatic compounds for MALDI-TOF MS were prepared. The formation of ionic liquids was proven by (1) H NMR spectroscopy. MALDI-Imaging mass spectrometry was applied to monitor the homogeneity. Our results show a superior sample spot homogeneity using ionic liquid matrices. Spots could be sampled several times without visible differences in the mass spectra. A frequently observed loss of matrix in the mass spectrometer vacuum was not observed. The necessary laser irradiance was reduced, which resulted in less polymer fragmentation. Ionic liquid matrices can be used to overcome segregation, a typical drawback of conventional MALDI dried-droplet preparations. Homogeneous sample spots are easy to prepare, stable in the MS vacuum and, thereby, improve the reproducibility of MALDI. Copyright © 2014 John Wiley & Sons, Ltd.

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

    Science.gov (United States)

    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.

  8. Evaluation of spot and passive sampling for monitoring, flux estimation and risk assessment of pesticides within the constraints of a typical regulatory monitoring scheme.

    Science.gov (United States)

    Zhang, Zulin; Troldborg, Mads; Yates, Kyari; Osprey, Mark; Kerr, Christine; Hallett, Paul D; Baggaley, Nikki; Rhind, Stewart M; Dawson, Julian J C; Hough, Rupert L

    2016-11-01

    In many agricultural catchments of Europe and North America, pesticides occur at generally low concentrations with significant temporal variation. This poses several challenges for both monitoring and understanding ecological risks/impacts of these chemicals. This study aimed to compare the performance of passive and spot sampling strategies given the constraints of typical regulatory monitoring. Nine pesticides were investigated in a river currently undergoing regulatory monitoring (River Ugie, Scotland). Within this regulatory framework, spot and passive sampling were undertaken to understand spatiotemporal occurrence, mass loads and ecological risks. All the target pesticides were detected in water by both sampling strategies. Chlorotoluron was observed to be the dominant pesticide by both spot (maximum: 111.8ng/l, mean: 9.35ng/l) and passive sampling (maximum: 39.24ng/l, mean: 4.76ng/l). The annual pesticide loads were estimated to be 2735g and 1837g based on the spot and passive sampling data, respectively. The spatiotemporal trend suggested that agricultural activities were the primary source of the compounds with variability in loads explained in large by timing of pesticide applications and rainfall. The risk assessment showed chlorotoluron and chlorpyrifos posed the highest ecological risks with 23% of the chlorotoluron spot samples and 36% of the chlorpyrifos passive samples resulting in a Risk Quotient greater than 0.1. This suggests that mitigation measures might need to be taken to reduce the input of pesticides into the river. The overall comparison of the two sampling strategies supported the hypothesis that passive sampling tends to integrate the contaminants over a period of exposure and allows quantification of contamination at low concentration. The results suggested that within a regulatory monitoring context passive sampling was more suitable for flux estimation and risk assessment of trace contaminants which cannot be diagnosed by spot

  9. Does Vitamin D Supplementation Enhance Musculoskeletal Performance in Individuals Identified as Vitamin D Deficient through Blood Spot Testing?

    Science.gov (United States)

    Murphy, Kellie A.

    This thesis investigated possible changes in performance after one month of vitamin D supplementation in individuals found to be vitamin D deficient or insufficient through blood spot testing. Thirty-two males, ages 18-32, participated. Each subject visited the lab three times in one-month, completing four performance tests each session, including an isometric mid-thigh pull and a vertical jump on a force plate, a isometric 90-degree elbow flexion test using a load cell, and a psychomotor vigilance test on a palm pilot. The initial lab included blood spot tests to find vitamin D levels. In a single blind manner, 16 subjects were assigned vitamin D and 16 the placebo. Repeated measures ANOVA analysis did not reveal any main effects for time (F=2.626, p=0.364), treatment (vitamin D3 vs placebo; F=1.282, p=0.999), or interaction effects for treatment by time (F=0.304, p=0.999) for maximum force production during an isometric mid-thigh pull. Repeated measures ANOVA analysis did not reveal any main effects for time (F=1.323, p=0.999), treatment (vitamin D3 vs placebo; F=0.510, p=0.999), or interaction effects for treatment by time (F= 1.625, p=0.860) for rate of force production during a vertical jump. Repeated measures ANOVA analysis did not reveal any main effects for time (F=0.194, p=0.999), treatment (vitamin D3 vs placebo; F=2.452, p=0.513), or interaction effects for treatment by time (F= 1.179, p=0.999) for maximal force production during a 90-degree isometric elbow flexion. Repeated measures ANOVA analysis did not reveal any main effects for time (F=1.710, p=0.804), treatment (vitamin D3 vs placebo; F=1.471, p=0.94), or interaction effects for treatment by time (F= 0.293, p=0.999) for mean reaction time to random stimuli during the psychomotor vigilance test. Repeated measures ANOVA analysis did not reveal any main effects for time (F=0.530, p=0.999), treatment (vitamin D3 vs placebo; F=0.141, p=0.999), or interaction effects for treatment by time (F=0.784 p=0

  10. Improved recovery of repeat intoxicated drivers using fingernails and blood spots to monitor alcohol and other substance abuse.

    Science.gov (United States)

    Bean, Pamela; Brown, Guida; Hallinan, Patricia; Becerra, Sergio; Lewis, Doug

    2017-01-02

    This study reports the results of a pilot program in Kenosha County that used a combination of direct biomarkers extracted from blood spots and nails to monitor repeat intoxicated drivers for their use of alcohol and drugs with a detection window spanning from 3 weeks to several months. The objectives were to test whether the direct biomarkers phosphatidylethanol (PEth), ethylglucuronide (EtG), and 5 drug metabolites would (1) help assessors obtain a more objective evaluation of repeat offenders during the assessment interview, (2) allow for timely identification of relapses and improve classification of drivers into risk categories, and (3) predict recidivism by identifying offenders most likely to obtain a subsequent operating while intoxicated (OWI) offense within 4 years of enrollment in the program. All (N = 261) repeat offenders were tested using PEth obtained from blood spots and EtG obtained from fingernails; 159 participants were also tested for a 5 drugs of abuse nail panel. Drivers were tested immediately after the assessment interview (baseline) and at 3, 6, 9, and 12 months after baseline. Based on biomarker results and self-reports of abstinence, offenders were classified into different risk categories and required to follow specific testing timelines based on the program's decision tree. The baseline analysis shows that 60% of drivers tested positive for alcohol biomarkers (EtG, PEth, or both) at the assessment interview, with lower detection rates (0-11%) for the 5 drug metabolites. The comparison of biomarkers results to self-reports of abstinence identified 28% of all offenders as high risk and assigned them to more frequent testing and more intense monitoring. The longitudinal analysis shows that 56% (completers) of participants completed the program successfully and the remaining 44% (noncompliant) terminated prematurely. Two thirds (68%) of the completers were able to reduce or control their drinking and one third relapsed at least one time

  11. Penicillin Dried Blood Spot Assay for Use in Patients Receiving Intramuscular Benzathine Penicillin G and Other Penicillin Preparations To Prevent Rheumatic Fever.

    Science.gov (United States)

    Page-Sharp, Madhu; Coward, Jonathan; Moore, Brioni R; Salman, Sam; Marshall, Lewis; Davis, Timothy M E; Batty, Kevin T; Manning, Laurens

    2017-08-01

    Rheumatic heart disease (RHD) remains an important global health challenge. Administration of benzathine penicillin (BPG) every 3 to 4 weeks is recommended as a secondary prophylaxis to prevent recurrent episodes of acute rheumatic fever and subsequent RHD. Following intramuscular injection, BPG is hydrolyzed to penicillin G (benzylpenicillin). However, little is known of the pharmacokinetics (PK) of BPG in pediatric populations at high risk of RHD or of the pharmacokinetic-pharmacodynamic relationship between penicillin exposure and clinically relevant outcomes. Dried blood spot (DBS) assays can facilitate PK studies in situations where frequent venous blood sampling is logistically difficult. A liquid chromatography-mass spectroscopy assay for penicillin G in plasma and DBS was developed and validated. Application of the DBS assay for PK studies was confirmed using samples from adult patients receiving penicillin as part of an infection management plan. The limit of quantification for penicillin G in DBS was 0.005 mg/liter. Penicillin G is stable in DBS for approximately 12 h at room temperature (22°C), 6 days at 4°C, and >1 month at -20°C. Plasma and DBS penicillin G concentrations for patients receiving BPG and penicillin G given via bolus doses correlated well and had comparable time-concentration profiles. There was poor correlation for patients receiving penicillin via continuous infusions, perhaps as a result of the presence of residual penicillin in the peripherally inserted central catheter, from which the plasma samples were collected. The present DBS penicillin G assay can be used as a surrogate for plasma concentrations to provide valid PK data for studies of BPG and other penicillin preparations developed to prevent rheumatic fever and RHD. Copyright © 2017 American Society for Microbiology.

  12. [DNA quantification of blood samples pre-treated with pyramidon].

    Science.gov (United States)

    Zhu, Chuan-Hong; Zheng, Dao-Li; Ni, Rao-Zhi; Wang, Hai-Sheng; Ning, Ping; Fang, Hui; Liu, Yan

    2014-06-01

    To study DNA quantification and STR typing of samples pre-treated with pyramidon. The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.

  13. Development and optimization of an internally controlled dried blood spot assay for surveillance of human immunodeficiency virus type-1 drug resistance.

    Science.gov (United States)

    Buckton, Andrew J; Bissett, Sara L; Myers, Richard E; Beddows, Simon; Edwards, Simon; Cane, Patricia A; Pillay, Deenan

    2008-12-01

    We present the evaluation of a methodology for the genotypic assessment of human immunodeficiency virus type-1 (HIV-1) drug resistance, optimized for use with dried blood spots (DBS). The ability to generate HIV-1 protease (PR) and reverse transcriptase (RT) contiguous amplicons and nucleotide sequences from DBS was evaluated. Different collection matrices and extraction methodologies were compared. The relative subtype sensitivity of the amplification strategy was assessed using a comprehensive panel of plasmids representing A-H subtypes. A panel of DBS and plasma specimens was subjected to HIV genotyping. Sequences generated from each sample type were compared. Extensive replicate testing revealed most sensitivity with the use of 903 filter paper and silica/guanidine extraction, which had an estimated 95% inclusivity endpoint of 1542 proviral copies/mL, as compared with 21 573 proviral copies/mL for the FTA system. All HIV-1 group M subtypes analysed-with the exception of subtypes A2, AE, AG, F and H-had a relative sensitivity of /=1000 copies/mL were successfully amplified and sequenced. Twelve specimens had pol genotyping from both plasma and DBS samples. Sequence analysis and drug resistance interpretation revealed that 10 (83%) provided concordant drug resistance interpretation. Our results demonstrate that the technique is appropriate for surveillance of drug resistance in untreated individuals and those with virological failure on therapy.

  14. A Simple Phosphate-Buffered-Saline-Based Extraction Method Improves Specificity of HIV Viral Load Monitoring Using Dried Blood Spots.

    Science.gov (United States)

    Makadzange, A Tariro; Boyd, F Kathryn; Chimukangara, Benjamin; Masimirembwa, Collen; Katzenstein, David; Ndhlovu, Chiratidzo E

    2017-07-01

    Although Roche COBAS Ampliprep/COBAS TaqMan (CAP/CTM) systems are widely used in sub-Saharan Africa for early infant diagnosis of HIV from dried blood spots (DBS), viral load monitoring with this system is not practical due to nonspecific extraction of both cell-free and cell-associated viral nucleic acids. A simplified DBS extraction technique for cell-free virus elution using phosphate-buffered saline (PBS) may provide an alternative analyte for lower-cost quantitative HIV virus load (VL) testing to monitor antiretroviral therapy (ART). We evaluated the CAP/CTM v2.0 assay in 272 paired plasma and DBS specimens using the cell-free virus elution method and determined the level of agreement, sensitivity, and specificity at thresholds of target not detected (TND), target below the limit of quantification (BLQ) (1,000 copies/ml, the sensitivities, specificities, positive predictive values (PPV), and negative predictive values (NPV) were 92.7%, 100%, 100%, and 94.3%, respectively. PBS elution of DBS offers a sensitive and specific method for monitoring plasma viremia among adults and children on ART at the WHO-recommended threshold of >1,000 copies/ml on the Roche CAP/CTM system. Copyright © 2017 Makadzange et al.

  15. Pyrosequencing dried blood spots reveals differences in HIV drug resistance between treatment naïve and experienced patients.

    Directory of Open Access Journals (Sweden)

    Hezhao Ji

    Full Text Available Dried blood spots (DBS are an alternative specimen collection format for HIV-1 genotyping. DBS produce HIV genotyping results that are robust and equivalent to plasma when using conventional sequencing methods. However, using tagged, pooled pyrosequencing, we demonstrate that concordance between plasma and DBS is not absolute and varies according to viral load (VL, duration of HIV infection and antiretroviral therapy (ART status. The plasma/DBS concordance is the highest when VL is ≥5,000 copies/ml and/or the patient has no ART exposure and/or when the duration of HIV infection is ≤2 years. Stepwise regression analysis revealed that VL is most important independent predictor for concordance of DBS with plasma genotypes. This is the first study to use next generation sequencing to identify discordance between DBS and plasma genotypes. Consideration should be given to VL, duration of infection, and ART exposure when interpreting DBS genotypes produced using next generation sequencing. These findings are of particular significance when DBS are to be used for clinical monitoring purposes.

  16. Validation of amino-acids measurement in dried blood spot by FIA-MS/MS for PKU management.

    Science.gov (United States)

    Bruno, C; Dufour-Rainfray, D; Patin, F; Vourc'h, P; Guilloteau, D; Maillot, F; Labarthe, F; Tardieu, M; Andres, C R; Emond, P; Blasco, H

    2016-09-01

    Phenylketonuria (PKU) is a metabolic disorder leading to high concentrations of phenylalanine (Phe) and low concentrations of tyrosine (Tyr) in blood and brain that may be neurotoxic. This disease requires a regular monitoring of plasma Phe and Tyr as well as branched-chain amino-acids concentrations to adapt the Phe-restricted diet and other therapy that may be prescribed in PKU. We validated a Flow Injection Analysis tandem Mass Spectrometry (FIA-MS/MS) to replace the enzymatic method routinely used for neonatal screening in order to monitor in parallel to Phe, Tyr and branched-chain amino-acids not detected by the enzymatic method. We ascertained the performances of the method: linearity, detection and quantification limits, contamination index, accuracy. We cross validated the FIA-MS/MS and enzymatic methods and we evaluated our own reference ranges to monitor Phe, Tyr, Leu, Val on 59 dried blood spots of normal controls. We also evaluated Tyr, Leu and Val concentrations in PKU patients to detect some potential abnormalities, not evaluated by the enzymatic method. We developed a rapid method with excellent performances including precision and accuracy PKU patients had lower concentrations of Tyr, Leu and/or Val that could not be detected by the enzymatic method. Based on laboratory accreditation recommendations, we validated a robust, rapid and reliable FIA-MS/MS method to monitor plasma Phe concentrations but also Tyr, Leu and Val concentrations, suitable for PKU management. We evaluated our own reference ranges of concentration for a routine application of this method. Copyright © 2016 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  17. Blood spot-based measures of glucose homeostasis and diabetes prevalence in a nationally representative population of young US adults.

    Science.gov (United States)

    Nguyen, Quynh C; Whitsel, Eric A; Tabor, Joyce W; Cuthbertson, Carmen C; Wener, Mark H; Potter, Alan J; Halpern, Carolyn T; Killeya-Jones, Ley A; Hussey, Jon M; Suchindran, Chirayath; Harris, Kathleen Mullan

    2014-12-01

    We investigated understudied biomarker-based diabetes among young US adults, traditionally characterized by low cardiovascular disease risk. We examined 15,701 participants aged 24 to 32 years at Wave IV of the National Longitudinal Study of Adolescent Health (Add Health, 2008). The study used innovative and relatively noninvasive methods to collect capillary whole blood via finger prick at in-home examinations in all 50 states. Assays of dried blood spots produced reliable and accurate values of HbA1c. Reliability was lower for fasting glucose and lowest for random glucose. Mean (SD) HbA1c was 5.6% (0.8%). More than a quarter (27.4%) had HbA1c-defined prediabetes. HbA1c was highest in the black, non-Hispanic race/ethnic group, inversely associated with education, and more common among the overweight/obese and physically inactive. The prevalence of diabetes defined by previous diagnosis or use of antidiabetic medication was 2.9%. Further incorporating HbA1c and glucose values, the prevalence increased to 6.8%, and among these participants, 38.9% had a previous diagnosis of diabetes (i.e., aware). Among those aware, 37.6% were treated and 64.0% were controlled (i.e., HbA1c young adults faces a historically high risk of diabetes but there is ample opportunity for early detection and intervention. Copyright © 2014 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    ... HUMAN SERVICES National Institutes of Health Donating Your Blood, Tissue, and Other Samples You have the choice to donate samples, such as blood and tissue, for medical research. Medical researchers use samples to ...

  20. Two novel nonradioactive polymerase chain reaction-based assays of dried blood spots, genomic DNA, or whole cells for fast, reliable detection of Z and S mutations in the alpha 1-antitrypsin gene

    DEFF Research Database (Denmark)

    Andresen, B S; Knudsen, I; Jensen, P K

    1992-01-01

    from individuals who are normal, heterozygous, or homozygous for the mutations. We show that the two assays can be performed with purified genomic DNA as well as with boiled blood spots. The new assays were validated by parallel testing with a technique in which PCR is combined with allele......-specific oligonucleotide (ASO) probes. In all cases tested the results obtained by the different techniques were in accordance. The new assays can be used for prenatal diagnostics and can be performed directly with boiled tissue samples. Because the new assays are easy to perform and reliable, we conclude...

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

    Science.gov (United States)

    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.

  2. Ambulatory blood pressure and blood lipids in a multiethnic sample of healthy adults.

    Science.gov (United States)

    James, Gary D; Van Berge-Landry, Helene M; Morrison, Lynn A; Reza, Angela M; Nicolaisen, Nicola M; Bindon, James R; Brown, Daniel E

    2013-01-01

    Elevated blood pressure (BP), elevated serum cholesterol, and aberrant lipoprotein fractions (low levels of high-density lipoprotein (HDL) and high levels of low-density lipoprotein fractions and triglycerides) have all been used as measures that assess the "metabolic syndrome" and more recently in indexes of allostatic load, which are designed to assess the degree of integrated metabolic pathology. While there are ample data regarding the interrelationships of these measures in various pathophysiological settings, there are limited data regarding the interrelationship of ambulatory BP (ABP) and blood lipids in healthy subjects. The present study evaluates ABP-blood lipid relationships in a multiethnic sample of healthy adults. The subjects were 37 men (age = 40.9 ± 10.7 years) and 42 women (age = 35.8 ± 10.4 years) who were employed as hotel workers in Hawaii. Each wore an ABP monitor for one midweek workday and had pressures averaged in three daily microenvironments (work, home, and during sleep). They also had fasting blood samples taken for lipid profiling. Multivariate analysis of covariance shows that there was a strong inverse relationship between HDL and both systolic (P act as a group in healthy adults but that higher HDL is associated with lower BP. This latter finding is consistent with research that shows that HDL promotes vasodilation via its effect on endothelial nitric oxide synthase. Copyright © 2013 Wiley Periodicals, Inc.

  3. Validation and Assessment of Three Methods to Estimate 24-h Urinary Sodium Excretion from Spot Urine Samples in High-Risk Elder Patients of Stroke from the Rural Areas of Shaanxi Province

    Directory of Open Access Journals (Sweden)

    Wenxia Ma

    2017-10-01

    Full Text Available Background: 24-h urine collection is regarded as the “gold standard” for monitoring sodium intake at the population level, but ensuring high quality urine samples is difficult to achieve. The Kawasaki, International Study of Sodium, Potassium, and Blood Pressure (INTERSALT and Tanaka methods have been used to estimate 24-h urinary sodium excretion from spot urine samples in some countries, but few studies have been performed to compare and validate these methods in the Chinese population. Objective: To compare and validate the Kawasaki, INTERSALT and Tanaka formulas in predicting 24-h urinary sodium excretion using spot urine samples in 365 high-risk elder patients of strokefrom the rural areas of Shaanxi province. Methods: Data were collected from a sub-sample of theSalt Substitute and Stroke Study. 365 high-risk elder patients of stroke from the rural areas of Shaanxi province participated and their spot and 24-h urine specimens were collected. The concentrations of sodium, potassium and creatinine in spot and 24-h urine samples wereanalysed. Estimated 24-h sodium excretion was predicted from spot urine concentration using the Kawasaki, INTERSALT, and Tanaka formulas. Pearson correlation coefficients and agreement by Bland-Altman method were computed for estimated and measured 24-h urinary sodium excretion. Results: The average 24-h urinary sodium excretion was 162.0 mmol/day, which representing a salt intake of 9.5 g/day. Three predictive equations had low correlation with the measured 24-h sodium excretion (r = 0.38, p < 0.01; ICC = 0.38, p < 0.01 for the Kawasaki; r = 0.35, p < 0.01; ICC = 0.31, p < 0.01 for the INTERSALT; r = 0.37, p < 0.01; ICC = 0.34, p < 0.01 for the Tanaka. Significant biases between estimated and measured 24-h sodium excretion were observed (all p < 0.01 for three methods. Among the three methods, the Kawasaki method was the least biased compared with the other two methods (mean bias: 31.90, 95% Cl: 23.84, 39

  4. Comparison of Measurements of Autoantibodies to Glutamic Acid Decarboxylase and Islet Antigen-2 in Whole Blood Eluates from Dried Blood Spots Using the RSR-Enzyme Linked Immunosorbent Assay Kits and In-House Radioimmunoassays

    Directory of Open Access Journals (Sweden)

    Anders Persson

    2010-01-01

    Full Text Available To evaluate the performance of dried blood spots (DBSs with subsequent analyses of glutamic acid decarboxylase (GADA and islet antigen-2 (IA-2A with the RSR-ELISAs, we selected 80 children newly diagnosed with type 1 diabetes and 120 healthy women. DBSs from patients and controls were used for RSR-ELISAs while patients samples were analysed also with in-house RIAs. The RSR-ELISA-GADA performed well with a specificity of 100%, albeit sensitivity (46% was lower compared to in RIA (56%; P=.008. No prozone effect was observed after dilution of discrepant samples. RSR-ELISA-IA-2A achieved specificity of 69% and sensitivity was lower (59% compared with RIA (66%; P<.001. Negative or low positive patients and control samples in the RSR-ELISA-IA-2A increased after dilution. Eluates from DBS can readily be used to analyse GADA with the RSR-ELISA, even if low levels of autoantibodies were not detected. Some factor could disturb RSR-ELISA-IA-2A analyses.

  5. Characteristics of a new fully programmable blood sampling device for monitoring blood radioactivity during PET

    International Nuclear Information System (INIS)

    Boellaard, R.; Lingen, A. van; Balen, S.C.M. van; Hoving, B.G.; Lammertsma, A.A.

    2001-01-01

    The first performance tests of a new fully programmable blood sampling device for monitoring blood radioactivity during positron emission tomography (PET) are described. Blood is withdrawn through 1-mm internal diameter tubing using an infusion pump which can be operated at rates varying from 0 to 600 ml/h. Activity in blood is measured by a 6-cm-thick bismuth germanate crystal connected to a photomultiplier tube and multichannel analyser (MCA) which are positioned within 6 cm lead shielding. Positioning of the tubing is an exact and simple procedure. The minimal readout time of the MCA is 1 s. Two independent energy windows can be set. Operation of the pump and MCA is fully controlled by a PC, i.e. sampling time, interval time and pump rate can be varied at any time during the PET scan by user-defined scripts. A number of characteristics of the new system were studied, such as sensitivity, dead time, linearity, effect of background radiation and pump rate as a function of input pressure. In addition, dispersion was measured as a function of pump rate. Finally, first clinical results were compared with manual samples. The sensitivity equalled 0.7 and 0.2 cps/Bq for 511- and 1022-keV 30% energy windows, respectively, and the system dead time was 500 ns. The system remained linear within 2% with activity concentrations up to 2.5 MBq/cc. Short-term reproducibility was better than 3% for a 1-h period. Long-term reproducibility was about 5% (1SD), which was mainly caused by variation in the diameter of the tubing. If the device was positioned in such a way that maximum shielding was directed towards the patient, the effects of background radiation from the patient on the measured activity concentration for clinically relevant conditions was minimal ( -1 were observed for pump rates higher than 300 ml/h, indicating that the system dispersion is small. Clinical data showed an excellent agreement to within 3% (1SD) between the results obtained with the new system and

  6. Real-time polymerase chain reaction optimised for hepatitis C virus detection in dried blood spots from HIV-exposed infants, KwaZulu-Natal, South Africa

    OpenAIRE

    Anneta Naidoo; Raveen Parboosing; Pravi Moodley

    2016-01-01

    Background: There is a paucity of data on the prevalence of hepatitis C virus (HCV) in children, particularly in sub-Saharan Africa. A major obstacle in resource-limited settings for polymerase chain reaction (PCR) testing is the necessity for specimen transportation and storage at low temperatures. There are numerous recent studies of using real-time HCV PCR for diagnosis and screening of plasma and serum, but few have looked at using dried blood spot (DBS) specimens. Objectives: The aim ...

  7. Next-generation sequencing of dried blood spot specimens: a novel approach to HIV drug-resistance surveillance.

    Science.gov (United States)

    Ji, Hezhao; Li, Yang; Graham, Morag; Liang, Ben Binhua; Pilon, Richard; Tyson, Shari; Peters, Geoff; Tyler, Shaun; Merks, Harriet; Bertagnolio, Silvia; Soto-Ramirez, Luis; Sandstrom, Paul; Brooks, James

    2011-01-01

    HIV drug-resistance (DR) surveillance in resource-limited settings can be performed using dried blood spots (DBS) because of ease of collection, transportation and storage. Analysis of pooled specimens on next-generation sequencing (NGS)-based platforms, such as the 454 pyrosequencing, is an efficient sequencing method for determining HIV DR rates. In this study, we conducted HIV DR surveillance on DBS using NGS and identified minority variants in individual patients. A total of 48 extracts of DBS from an HIV DR surveillance study in Mexico City were re-amplified using primers tagged with multiplex identifiers, pooled and pyrosequenced. Consensus sequences were generated for each specimen with mixtures identified at positions where >20% of the reads contained a variant. Individual consensus sequences were then analysed for DR mutations and compared with those derived from Sanger sequencing. DBS analysed with tagged pooled pyrosequencing (TPP) were highly concordant with Sanger sequencing genotypes from matching plasma and DBS (99.21% and 99.51%, respectively). An exception was an M184I mutation only detected with TPP of DBS at a frequency of 20.4%. Multiple specimens had minority variant reads below the 20% mixture threshold. TPP using DBS is an effective method for HIV DR surveillance. TPP for genotyping results in cost savings of 40% over conventional in-house methods. The effect of low-abundance DR mutations, undetectable by conventional methods, remains to be determined. This technology might be applied to any HIV specimen (plasma/serum) and can also be used for other diagnostic assays where DNA sequencing is required. © 2011 International Medical Press

  8. Newborn Dried Blood Spot Polymerase Chain Reaction to Identify Infants with Congenital Cytomegalovirus-Associated Sensorineural Hearing Loss.

    Science.gov (United States)

    Ross, Shannon A; Ahmed, Amina; Palmer, April L; Michaels, Marian G; Sánchez, Pablo J; Stewart, Audra; Bernstein, David I; Feja, Kristina; Fowler, Karen B; Boppana, Suresh B

    2017-05-01

    To determine the utility of dried blood spot (DBS) polymerase chain reaction (PCR) in identifying infants with cytomegalovirus (CMV) infection-associated sensorineural hearing loss (SNHL). Newborns at 7 US hospitals between March 2007 and March 2012 were screened for CMV by saliva rapid culture and/or PCR. Infected infants were monitored for SNHL during the first 4 years of life to determine sensitivity, specificity, and positive and negative likelihood ratios of DBS PCR for identifying CMV-associated SNHL. DBS at birth was positive in 11 of 26 children (42%) with SNHL at age 4 years and in 72 of 270 children (27%) with normal hearing (P = .11). The sensitivity (42.3%; 95% CI, 23.4%-63.1%) and specificity (73.3%; 95% CI, 67.6%-78.5%) was low for DBS PCR in identifying children with SNHL at age 4 years. The positive and negative likelihood ratios of DBS PCR positivity to detect CMV-associated SNHL at age 4 years were 1.6 (95% CI, 0.97-2.6) and 0.8 (95% CI, 0.6-1.1), respectively. There was no difference in DBS viral loads between children with SNHL and those without SNHL. DBS PCR for CMV has low sensitivity and specificity for identifying infants with CMV-associated hearing loss. These findings, together with previous reports, demonstrate that DBS PCR does not identify either the majority of CMV-infected newborns or those with CMV-associated SNHL early in life. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Analysis of the GH content within archived dried blood spots of newborn screening cards from children diagnosed with growth hormone deficiency after the neonatal period.

    Science.gov (United States)

    Binder, G; Hettmann, S; Weber, K; Kohlmüller, D; Schweizer, R

    2011-12-01

    It is unknown whether GH secretion of children with growth hormone deficiency (GHD) is already diminished at birth. We aimed to determine the GH content within archived dried blood spots of newborn screening cards from children diagnosed with GHD at childhood. At our hospital, all children with the diagnosis of GHD and an actual age GH content. Reference values were calculated based on 600 anonymous newborn screening cards of different ages. Median GH content within the archived dried blood spots of the reference had declined by 30% during the first year and by further 35% during the next 8.5years of storage. After correction for time of storage, four out of the 16 archived dried blood spots of the GHD children contained low amounts of GH (≤5th percentile). Diminished GH secretion at birth was absent in isolated GHD, but associated with multiple pituitary hormone deficiency (MPHD) (P=0.0013), ectopic neurohypophysis (P=0.0013), lower GH test peak values (P=0.02) and higher weight at diagnosis (P=0.015). Children with isolated GHD have normal GH secretory capacity during the first week of life while the majority of children with MPHD and pituitary malformation were GH deficient immediately after birth. Copyright © 2011 Elsevier Ltd. All rights reserved.

  10. Quantification of the 3α and 3β epimers of 25-hydroxyvitamin D3in dried blood spots by LC-MS/MS using artificial whole blood calibration and chemical derivatization.

    Science.gov (United States)

    Müller, Miriam J; Stokes, Caroline S; Volmer, Dietrich A

    2017-04-01

    While the biological function of the 3α epimer of 25-hydroxyvitamin D 3 (25(OH)D 3 ) remains unknown, its presence needs to be accurately captured and separated from the main 3β epimer, to avoid positive bias in vitamin D status analyses. Several recent LC-MS/MS assays for 25(OH)D 3 successfully separate the 3α and 3β epimers by chromatography. Unfortunately, none of the existing LC-MS/MS assays, which utilize dried blood spots (DBS) as sampling/storage vessels, is able to quantify the individual epimers. DBS are often used for analysis of infant blood, however, and these samples are particularly likely to contain significant levels of interfering 3α epimer. Furthermore, proper calibration of DBS samples is much more difficult to achieve than for liquid serum or plasma samples. We addressed this important issue by creating an artificial vitamin D-free whole blood for calibration and then quantified 3α- and 3β-25(OH)D 3 levels from DBS. After chemical derivatization, the vitamin D epimers were separated on a PFP column and concentrations determined by electrospray ionization LC-MS/MS on a triple quadrupole mass spectrometer. Calibration with artificial whole blood showed improved precision over standard addition (7.6 versus 31.5% RSD for 3β-25(OH)D 3 ). The limits of quantification for 3β-25(OH)D 3 and for 3α-25(OH)D 3 were 1.0 and 0.1ng/mL, respectively. Excellent intra/interday precisions between 2.1 and 2.2% CV (intra) and 4.4-5.3% CV (inter) were established for 3β-25(OH)D 3 and 3α-25(OH)D 3 . For 3β-25(OH)D 3 , only small concentration-independent bias and deviation of <3.3ng/mL were seen between serum LC-MS/MS and DBS-LC-MS/MS measurements; analyses of 3α-25(OH)D 3 showed deviations of <0.8ng/mL in all experiments. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. What is the right blood hematocrit preparation procedure for standards and quality control samples for dried blood spot analysis?

    NARCIS (Netherlands)

    Koster, Remco A.; Alffenaar, Jan-Willem C.; Botma, Rixt; Greijdanus, Ben; Touw, Daan J.; Uges, Donald R. A.; Kosterink, Jos G. W.

    2015-01-01

    Remco Koster is a research analyst and PhD candidate at the University Medical Center Groningen and University of Groningen. He has been working in the field of bioanalysis for over 13 years, where he has developed numerous analytical methods using LC-MS/MS. His main research focus is the influence

  12. Automatic collection of bovine blood samples | Hale | South African ...

    African Journals Online (AJOL)

    A technique is described which allows automatic collection of jugular venous blood from tethered cows. In this system, blood is pumped continuously from an intravenous cannula which has a double lumen while an anticoagulant is pumped through the second opening. Diluted blood is collected in a fraction collector which ...

  13. The protein-creatinine ratio in spot morning urine samples and 24-h urinary protein excretion in patients with systemic lupus erythematosus.

    Science.gov (United States)

    Salesi, Mansour; Karimifar, Mansoor; Farajzadegan, Ziba; Esalatmanesh, Kamal; Khosravi, Shahrzad; Fallahi, Parvin; Akbarian, Mahmood

    2009-03-01

    A 24-h urinary protein is a standard way to diagnose lupus nephritis. Assessment of protein-creatinine (Pr-Cr) ratio in morning spot urine is a valuable method in diabetic patients but not use in systemic lupus erythematous (SLE) patients routinely. In this study Pr-Cr ratio in spot urine was compare with 24-h urine protein; if they have valuable correlation we can use this test instead of 24-h urinary protein. The aim of this study was to evaluate the correlation of spot urine Pr-Cr ratio for prediction of significant proteinuria (>or=300 mg/24 h) in patients with SLE. A cross-section study was conducted in 74 hospitalized women with SLE. The correlation between Pr-Cr in first morning urine specimens and urinary protein excretion in 24-h collections were analyzed. Correlation between Pr-Cr ratio in spot morning urine specimens and urinary protein excretion in 24-h collections was significant (P protein excretion and its measurement, enabling the methods to be used interchangeably creatinine ratio in spot morning urine samples is a precise indicator of proteinuria in patients with lupus nephritis and represents a simple and inexpensive procedure in establishing severity of proteinuria in patients with SLE.

  14. Effect of order of draw of blood samples during phlebotomy on routine biochemistry results.

    Science.gov (United States)

    Sulaiman, Raashda A; Cornes, Michael P; Whitehead, Simon J; Othonos, Nadia; Ford, Clare; Gama, Rousseau

    2011-11-01

    To investigate whether incorrect order of draw of blood samples during phlebotomy causes in vitro potassium ethylenediaminetetraacetic acid EDTA (kEDTA) contamination of blood samples. Serum kEDTA, potassium, calcium, magnesium, alkaline phosphatase, zinc and iron concentrations were measured in blood samples drawn before and after collecting blood into kEDTA containing sample tubes by an experienced phlebotomist using the Sarstedt Safety Monovette system. EDTA was undetectable in all samples. The concentrations of other analytes were similar in blood samples drawn before and after collection of the EDTA blood sample. Order of draw of blood samples using the Sarstedt Safety Monovette system has no effect on serum biochemistry results, when samples are taken by an experienced phlebotomist.

  15. Screening for adenylosuccinate lyase deficiency using tandem mass spectrometry analysis of succinylpurines in neonatal dried blood spots.

    Science.gov (United States)

    Zikanova, Marie; Krijt, Jakub; Skopova, Vaclava; Krijt, Matyas; Baresova, Veronika; Kmoch, Stanislav

    2015-01-01

    Stable isotope dilution coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) is the sensitive method for screening for various inherited metabolic disorders using dried blood spots (DBSs). We present a method for LC-MS/MS determination of succinyladenosine (SAdo) and succinylaminoimidazole carboxamide riboside (SAICAr), biomarkers for adenylosuccinate lyase deficiency (dADSL), in DBS. SAICAr and SAdo were separated on a Symmetry-C18 column and detected using positive electrospray ionisation in selected reaction monitoring mode. The quantification was performed using the isotopically labelled internal standards SAdo-(13)C4 and SAICAr-(13)C4, which were prepared via ADSL-catalysed reactions of fumarate-(13)C4 with adenosine monophosphate and aminoimidazole carboxamide ribotide, respectively, and subsequent alkaline phosphatase-catalysed dephosphorylation of the resulting products. The detection of SAICAr and SAdo in DBS was linear over the range of 0-25μmol/L. The respective intra-assay and inter-assay imprecision values were less than 10.7% and 15.2% for SAICAr and 4.7% and 5.7% for SAdo. The recoveries from DBS spiked with different concentrations of SAICAr and SAdo were between 94% and 117%. The concentrations of SAICAr and SAdo were higher in the archived DBS from dADSL patients (SAICAr, 0.03-4.7μmol/L; SAdo, 1.5-21.3μmol/L; n=5) compared to those of the control subjects (SAICAr, 0-0.026μmol/L; SAdo, 0.06-0.14μmol/L; n=31), even after DBSs from dADSL patients were stored for 2-23years. We developed and validated a method of succinylpurine analysis in DBS that improves selective screening for dADSL in the paediatric population and may be used for retrospective diagnosis to aid the genetic counselling of affected families. Copyright © 2014 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  16. Incident Infection and Resistance Mutation Analysis of Dried Blood Spots Collected in a Field Study of HIV Risk Groups, 2007-2010.

    Science.gov (United States)

    Wei, Xierong; Smith, Amanda J; Forrest, David W; Cardenas, Gabriel A; Beck, Dano W; LaLota, Marlene; Metsch, Lisa R; Sionean, Catlainn; Owen, S Michele; Johnson, Jeffrey A

    2016-01-01

    To assess the utility of cost-effective dried blood spot (DBS) field sampling for incidence and drug resistance surveillance of persons at high risk for HIV infection. We evaluated DBS collected in 2007-2010 in non-clinical settings by finger-stick from HIV-positive heterosexuals at increased risk of HIV infection (n = 124), men who have sex with men (MSM, n = 110), and persons who inject drugs (PWID, n = 58). Relative proportions of recent-infection findings among risk groups were assessed at avidity index (AI) cutoffs of ≤25%, ≤30%, and ≤35%, corresponding to an infection mean duration of recency (MDR) of 220.6, 250.4, and 278.3 days, respectively. Drug resistance mutation prevalence was compared among the risk groups and avidity indices. HIV antibody avidity testing of all self-reported ARV-naïve persons (n = 186) resulted in 9.7%, 11.3% and 14.0% with findings within the 221, 250, and 278-day MDRs, respectively. The proportion of ARV-naïve MSM, heterosexuals, and PWID reporting only one risk category who had findings below the suggested 30% AI was 23.1%, 6.9% and 3.6% (pinfection results than did heterosexuals and PWID. The disproportionately higher recent-infection findings for MSM as compared to PWID and heterosexuals increased as the MDR window increased. Unreported ARV use might explain greater recent-infection findings and drug resistance in this MSM population. DBS demonstrated utility in expanded HIV testing; however, optimal field handling is key to accurate recent-infection estimates.

  17. Bacterial Isolates from Blood Samples of Patients in University of ...

    African Journals Online (AJOL)

    The thioglychollate broth was sub cultured onto blood agar plate for anaerobic incubation, while the brain heart infusion broth was sub cultured onto chocolate, blood agar and McConkey agar for aerobic incubation. The anaerobic incubation was done by putting the culture plates into an anaerobic jar and filling it with ...

  18. High plasma corticosterone levels persist during frequent automatic blood sampling in rats

    DEFF Research Database (Denmark)

    Abelson, Klas S P; Adem, Bashir; Royo, Felix

    2005-01-01

    the importance of considering the frequency of blood withdrawal during automated blood sampling. This parameter may have an impact on the experimental results when using blood corticosterone levels as a stress marker, but also during any in vivo study where blood is collected, since high corticosterone levels...

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

    Directory of Open Access Journals (Sweden)

    Juana del Valle Mendoza

    Full Text Available 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.

  20. Estimasi Sintesis Protein Mikrobia Rumen Menggunakan Ekskresi Derivat Purin dalam Urin dengan Teknik Spot Sampling pada Kambing Bligon dan Kambing Kejobong

    Directory of Open Access Journals (Sweden)

    Dianestu Putra

    2016-11-01

    Full Text Available This study were aimed to determine the correlation between concentration of purine derivatives (PD in spot sample with PD total excretion in Bligon and Kejobong goats and determine the appropriate sampling time, in order to predicting microbial protein synthesis in both breeds. Six male Bligon goats and six male Kejobong goats with age range from 8 to 14 months and body weight from 16 to 21 kg were placed in metabolism cages. Peanut straw and water were given to both groups of goats through ad libitum feeding and drinking. The study was done in 14 days for adaptation, 3 days for collection. Sample of feeds, feed residues, and feces were collected and then analyzed to determine dry matter and organic matter content. Spot urine and the total daily urine samples were also collected. Samples collection of spot sampling technique was run by taking the urine periodically with 3 hours intervals at 24 hours. Urine samples were analyzed for the content of creatinine and PD which includes allantoin, uric acid, xanthine, and hypoxanthine. Data were tested for the correlation between concentration of PD spot urine sample with total PD daily excretion. It is known that the concentration of PD and creatinine (µmol/L for Bligon were 1,418.40 and 202.85 respectively, while for Kejobong were 1,547.40 and 219.68 respectively. Total excretion of PD, allantoin, uric acid, xanthyne and hypoxanthine and creatinine (µmol/W0,75/day for Bligon were 114.14, 95.86, 17.31, 0.97, and 16.40 respectively, with microbial protein synthesis efficiency was 4.61 g N/kg degraded of organic matter in rumen (DOMR. Total excretion of PD allantoin, uric acid, xanthyne and hypoxanthine and creatinine (µmol/W0,75/day for Kejobong were 180.18, 158.17, 20.60, 1.40, and 24.87 respectively, with microbial protein synthesis efficiency was 6.90 g N/kg DOMR. Based on this study also known that the best time for spot sampling to determine the total excretion of PD in Bligon was in the range

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

    Science.gov (United States)

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

    2015-03-01

    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. To investigate the effects of samples storage temperature and time delay on blood gases, bicarbonate and PH results in human arterial blood samples. 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. 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 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. 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.

  2. The quantitative regional cerebral blood flow measurement with autoradiography method using 123I-IMP SPECT. Evaluation of arterialized venous blood sampling as a substitute for arterial blood sampling

    International Nuclear Information System (INIS)

    Ohnishi, Takashi; Yano, Takao; Nakano, Shinichi; Jinnouchi, Seishi; Nagamachi, Shigeki; Flores, L. II; Nakahara, Hiroshi; Watanabe, Katsushi.

    1996-01-01

    The purpose of this study is validation of calibrating a standard input function in autoradiography (ARG) method by one point venous blood sampling as a substitute for that by one point arterial blood sampling. Ten and 20 minutes after intravenous constant infusion of 123 I-IMP, arterialized venous blood sampling from a dorsal vein were performed on 15 patients having ischemic cerebrovascular disease. And arterial blood sampling from radial artery was performed 10 min after 123 I-IMP infusion. The mean difference rates of integrated input function between calibrated standard input function by arterial blood sampling at 10 min and that by venous blood sampling were 4.1±3% and 9.3±5.4% at 10 and 20 min after 123 I-IMP infusion, respectively. The ratio of venous blood radioactivity to arterial blood radioactivity at 10 min after 123 I-IMP infusion was 0.96±0.02. There was an excellent correlation between ARG method CBF values obtained by arterial blood sampling at 10 min and those obtained by arterialized venous blood sampling at 10 min. In conclusion, a substitution by arterialized venous blood sampling from dorsal hand vein for artery can be possible. The optimized time for arterialized venous blood sampling was 10 min after 123 I-IMP infusion. (author)

  3. Blood samples in the neutron beam; Blutproben im Neutronenstrahl

    Energy Technology Data Exchange (ETDEWEB)

    Anon.

    2012-07-01

    Whether crocodile, platypus, man, or chicken: Always the hemoglobin in the red blood cells has the same task. It carries oxygen from the lungs throughout the body. In investigative manner and international team around Dr. Andreas Stadler from the Julic Research Center has deciphered in detail, how and why the hemoglobines of these creatures nevertheless differ. Their findings are among others interesting for the research on artificial blood.

  4. The impact of different blood sampling methods on laboratory rats under different types of anaesthesia

    DEFF Research Database (Denmark)

    Toft, Martin Fitzner; Petersen, Mikke Haxø; Dragsted, Nils

    2006-01-01

    Rats with implanted telemetry transponders were blood sampled by jugular puncture, periorbital puncture or tail vein puncture, or sampled by jugular puncture in carbon dioxide (CO?), isoflurane or without anaesthesia in a crossover design. Heart rate, blood pressure and body temperature were...... showed lower increases in blood pressure after, and fewer fluctuations in body temperature during sampling, and the post-anaesthetic effects of isoflurane, if any, seemed to disappear immediately after sampling. It is, therefore, concluded that blood sampling in rats by jugular puncture seems...

  5. A simple dried blood spot-method for in vivo measurement of ureagenesis by gas chromatography-mass spectrometry using stable isotopes.

    Science.gov (United States)

    Allegri, Gabriella; Deplazes, Sereina; Grisch-Chan, Hiu Man; Mathis, Déborah; Fingerhut, Ralph; Häberle, Johannes; Thöny, Beat

    2017-01-01

    Clinical management of inherited or acquired hyperammonemia depends mainly on the plasma ammonia level which is not a reliable indicator of urea cycle function as its concentrations largely fluctuate. The gold standard to assess ureagenesis in vivo is the use of stable isotopes. Here we developed and validated a simplified in vivo method with [ 15 N]ammonium chloride ([ 15 N]H 4 Cl) as a tracer. Non-labeled and [ 15 N]urea were quantified by GC-MS after extraction and silylation. Different matrices were evaluated for suitability of analysis. Ureagenesis was assessed in ornithine transcarbamylase (OTC)-deficient spf ash mice with compromised urea cycle function during fasted and non-fasted feeding states, and after rAAV2/8-vector delivery expressing the murine OTC-cDNA in liver. Blood (5μL) was collected through tail vein puncture before and after [ 15 N]H 4 Cl intraperitoneal injections over a two hour period. The tested matrices, blood, plasma and dried blood spots, can be used to quantify ureagenesis. Upon [ 15 N]H 4 Cl challenge, urea production in spf ash mice was reduced compared to wild-type and normalized following rAAV2/8-mediated gene therapeutic correction. The most significant difference in ureagenesis was at 30min after injection in untreated spf ash mice under fasting conditions (19% of wild-type). Five consecutive injections over a period of five weeks had no effect on body weight or ureagenesis. This method is simple, robust and with no apparent risk, offering a sensitive, minimal-invasive, and fast measurement of ureagenesis capacity using dried blood spots. The stable isotope-based quantification of ureagenesis can be applied for the efficacy-testing of novel molecular therapies. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Nanobubbles Form at Active Hydrophobic Spots on the Luminal Aspect of Blood Vessels: Consequences for Decompression Illness in Diving and Possible Implications for Autoimmune Disease—An Overview

    Directory of Open Access Journals (Sweden)

    Ran Arieli

    2017-08-01

    Full Text Available Decompression illness (DCI occurs following a reduction in ambient pressure. Decompression bubbles can expand and develop only from pre-existing gas micronuclei. The different hypotheses hitherto proposed regarding the nucleation and stabilization of gas micronuclei have never been validated. It is known that nanobubbles form spontaneously when a smooth hydrophobic surface is submerged in water containing dissolved gas. These nanobubbles may be the long sought-after gas micronuclei underlying decompression bubbles and DCI. We exposed hydrophobic and hydrophilic silicon wafers under water to hyperbaric pressure. After decompression, bubbles appeared on the hydrophobic but not the hydrophilic wafers. In a further series of experiments, we placed large ovine blood vessels in a cooled high pressure chamber at 1,000 kPa for about 20 h. Bubbles evolved at definite spots in all the types of blood vessels. These bubble-producing spots stained positive for lipids, and were henceforth termed “active hydrophobic spots” (AHS. The lung surfactant dipalmitoylphosphatidylcholine (DPPC, was found both in the plasma of the sheep and at the AHS. Bubbles detached from the blood vessel in pulsatile flow after reaching a mean diameter of ~1.0 mm. Bubble expansion was bi-phasic—a slow initiation phase which peaked 45 min after decompression, followed by fast diffusion-controlled growth. Many features of decompression from diving correlate with this finding of AHS on the blood vessels. (1 Variability between bubblers and non-bubblers. (2 An age-related effect and adaptation. (3 The increased risk of DCI on a second dive. (4 Symptoms of neurologic decompression sickness. (5 Preconditioning before a dive. (6 A bi-phasic mechanism of bubble expansion. (7 Increased bubble formation with depth. (8 Endothelial injury. (9 The presence of endothelial microparticles. Finally, constant contact between nanobubbles and plasma may result in distortion of proteins and their

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

    Science.gov (United States)

    Mullins, Garrett R; Bruns, David E

    2017-10-01

    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 (pblood sample prevented bubble formation and fully protected the blood from PTS-induced hemolysis (pblood developed less foaming during PTS transport and was partially protected from hemolysis vs anticoagulated blood as indicated by lower LD (psample transport. Prevention of air bubble formation in blood samples during PTS transport protects samples from hemolysis. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. ABO and D typing and alloantibody screening in marrow samples: relevance to intraosseous blood transfusion.

    Science.gov (United States)

    Bäckman, Sari; Ångerman-Haasmaa, Susanne; Jousi, Milla; Siitonen, Sanna; Salmela, Katja

    2018-03-01

    Blood transfusion through the intraosseous route is gaining popularity in emergency medicine. Pretransfusion peripheral blood (PB) samples are usually not available in these patients, leading to discrepancies in blood group typing and a possible delay in transferring to group-specific blood products. The aim of this study was to assess the feasibility of ABO and D typing and red blood cell alloantibody screening in marrow (BM) samples. Direct and reverse ABO typing, D typing, and a two-cell alloantibody screen were performed in EDTA-anticoagulated BM samples with standard manual column agglutination techniques. EDTA-anticoagulated PB samples were used as controls. The mean age of the study subjects (n = 71) was 47 years (range, 1-82 years). All ABO groups and both D+ and D- types were represented. In all subjects, concordant results were observed for all analyses in BM and PB samples. In 15 (21%) of the samples, a discrepancy of one reaction strength step (1+) was observed in at least one of the analyses (Cohen's weighted κ = 0.993); this did not affect interpretation of the results. Blood group typing and alloantibody screening are feasible in BM samples, providing proof-of-concept that intraosseous samples for blood group serologic analyses can be collected from emergency patients before intraosseous blood transfusion. This will enable a timely transfer to group-specific blood products and enable conservation of the valuable universal-donor blood products. © 2018 AABB.

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

    Science.gov (United States)

    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.

  10. Effects of Blood Sample Collection Pre- and Post- Slaughter, Edta ...

    African Journals Online (AJOL)

    The samples were immediately subjected to Wet mount (WM), Haemotocrit centrifugation test (HCT) and Thin smear (TS) tests. The results revealed that, of the 100 samples examined, 19 (19%) were positive for the presence of Microfilaria spp while 6(6%) yielded Trypanosome spp. Of the 19 samples detected having ...

  11. Forensic Identification of Human Blood: comparison of two one-step presumptive tests for blood screening of crime scene samples.

    Directory of Open Access Journals (Sweden)

    Ana Flávia Belchior Andrade

    2014-08-01

    Full Text Available Blood is the most common body fluid found at crime scenes. One-step presumptive tests have been designed as a rapid immunological test for the qualitative detection of human hemoglobin in stool samples (faecal occult blood their usefulness for forensic purposes has been demonstrated before. In this study we compare Hexagon OBTI kit and FOB One-step Bioeasy kit sensitivity in the analysis of diluted blood samples. With Hexagon OBTI, positive test results are achieved in whole blood dilutions up to 1:1.000. Sensitivity decreased with aged samples, if samples were not stored under low temperatures regardless of which presumptive test is used. Whole blood tests must take into consideration that “hook” effect may interfere. Comparing both tests, OBTI Hexagon Kit is more sensible to detect diluted blood, showing a wider detection window in all conditions. This is interesting when analyzing forensic samples as forensic analysts usually do not know about the history of the analyzed sample before its collection.

  12. Validation of curve-fitting method for blood retention of 99mTc-GSA. Comparison with blood sampling method

    International Nuclear Information System (INIS)

    Ha-Kawa, Sang Kil; Suga, Yutaka; Kouda, Katsuyasu; Ikeda, Koshi; Tanaka, Yoshimasa

    1997-01-01

    We investigated a curve-fitting method for the rate of blood retention of 99m Tc-galactosyl serum albumin (GSA) as a substitute for the blood sampling method. Seven healthy volunteers and 27 patients with liver disease underwent 99m Tc-GSA scanning. After normalization of the y-intercept as 100 percent, a biexponential regression curve for the precordial time-activity curve provided the percent injected dose (%ID) of 99m Tc-GSA in the blood without blood sampling. The discrepancy between %ID obtained by the curve-fitting method and that by the multiple blood samples was minimal in normal volunteers 3.1±2.1% (mean±standard deviation, n=77 sampling). Slightly greater discrepancy was observed in patients with liver disease (7.5±6.1%, n=135 sampling). The %ID at 15 min after injection obtained from the fitted curve was significantly greater in patients with liver cirrhosis than in the controls (53.2±11.6%, n=13; vs. 31.9±2.8%, n=7, p 99m Tc-GSA and the plasma retention rate for indocyanine green (r=-0.869, p 99m Tc-GSA and could be a substitute for the blood sampling method. (author)

  13. Continuous quality control of the blood sampling procedure using a structured observation scheme

    DEFF Research Database (Denmark)

    Seemann, T. L.; Nybo, M.

    2015-01-01

    Background: An important preanalytical factor is the blood sampling procedure and its adherence to the guidelines, i.e. CLSI and ISO 15189, in order to ensure a consistent quality of the blood collection. Therefore, it is critically important to introduce quality control on this part of the process....... As suggested by the EFLM working group on the preanalytical phase we introduced continuous quality control of the blood sampling procedure using a structured observation scheme to monitor the quality of blood sampling performed on an everyday basis. Materials and methods: Based on our own routines the EFLM....... Conclusion: It is possible to establish a continuous quality control on blood sampling. It has been well accepted by the staff and we have already been able to identify critical areas in the sampling process. We find that continuous auditing increase focus on the quality of blood collection which ensures...

  14. The impact of different blood sampling methods on laboratory rats under different types of anaesthesia

    DEFF Research Database (Denmark)

    Toft, Martin Fitzner; Petersen, Mikke Haxø; Dragsted, Nils

    2006-01-01

    Rats with implanted telemetry transponders were blood sampled by jugular puncture, periorbital puncture or tail vein puncture, or sampled by jugular puncture in carbon dioxide (CO?), isoflurane or without anaesthesia in a crossover design. Heart rate, blood pressure and body temperature were...... for rats sampled from the tail vein, which showed fluctuations in body temperature in excess of 30 h after sampling. Increases in heart rate and blood pressure within the first hours after sampling indicated that periorbital puncture was the method that had the largest acute impact on the rats...... registered for three days after sampling. Initially blood pressure increased, but shortly after sampling it decreased, which led to increased heart rate. Sampling induced rapid fluctuations in body temperature, and an increase in body temperature. Generally, rats recovered from sampling within 2-3 h, except...

  15. Validation of a less invasive blood sampling technique in rabies serology using reduviid bugs (Triatominae, Hemiptera).

    Science.gov (United States)

    Vos, Ad C; Müller, Thomas; Neubert, Larissa; Voigt, Christian C

    2010-03-01

    During serologic rabies surveys, bleeding is often difficult or almost impossible in small or endangered mammals such as bats. Therefore, the usefulness of an alternative, less invasive technique--that is, the use of blood-sucking reduviid bugs (Dipetalogaster maximus and Rhodnius prolixus)--was investigated. Bugs were used in combination with a conventional method (retro-orbitale bleeding) to obtain blood samples from the same individual NMRI-mice (Mus musculus) vaccinated against rabies. Rabies virus-neutralizing antibody (VNA) titers between paired blood samples obtained from the same mice were compared. The accuracy (between-method comparison), precision (repeatability of results), and robustness (influence of digestion on blood parameter) of the bug method was evaluated. VNA titers in the blood sample obtained from the bugs' crops were equivalent to those from samples collected directly from the mice. No differences between samples taken from different bugs that had fed on the same mouse were noted. In addition, there were no changes in VNA titers in blood samples collected from the triatomine bugs for up to 4 hr after completion of the blood meal. This study demonstrates that the application of blood-sucking bugs offers a validated alternative for obtaining blood samples to determine rabies virus-neutralizing antibody titers and is highly suitable for animals with limited or no accessibility of veins by conventional sampling techniques.

  16. Detection of Salmonella typhi by nested polymerase chain reaction in blood, urine, and stool samples

    NARCIS (Netherlands)

    Hatta, Mochammad; Smits, Henk L.

    2007-01-01

    A nested polymerase chain reaction (PCR) specific for Salmonella enterica serovar Typhi was used for the detection of the pathogen in blood, urine, and stool samples from 131 patients with clinical suspicion of typhoid fever. The sensitivity of blood culture, the PCRs with blood, urine, and feces,

  17. Correlation between dried blood spot thin layer chromatography and plasma high performance liquid chromatography of leucine/isoleucine levels among Filipino patients with maple syrup urine disease (MSUD) seen at the Institute of Human Genetics, National Institutes of Health

    International Nuclear Information System (INIS)

    Yaplito-Lee, Joy; Chiong, Mary Anne D.; Rana, Michelle D.; Rama, Kahlil Izza D.; David-Padilla, Carmencita; Cavan, Barbra Charina; Cordero, Cynthia P.

    2008-01-01

    Management of patients with maple syrup urine disease (MSUD) includes a low protein diet, supplemented with special formulas and constant monitoring of branched chain amino acids (BCAA). The gold standard for monitoring BCAA is plasma amino acid analysis using high performance liquid chromatography (HPLC). In a developing country like the Philippines, however, the cost of this test is prohibitive to the majority of the patients. In our center, dried blood spot leucine/isoleucine (leu/ile) levels analysed by thin layer chromatography (TLC) is often used to diagnose and monitor these patients. This study was done to determine the correlation of leu/ile levels using the two methods (TLC and HPLC). A total of 46 MSUD patients were referred to the Biochemical Genetics Laboratory of the Institute of Human Genetics (IHG) from July 2001 to January 2004. Thirty five samples were obtained from 18 of these patients (some patients were seen at IHG more than once), and paired determinations of plasma amino acid using TLC and HPLC were made. The remaining samples were either hemolyzed or were not analyzed. The correlation coefficient [rho denoted as ρ] was estimated at a 95% confidence level using the Fisher's Z transformation. Of the 18 patients, 12 were males. The youngest was 1 day old and the oldest was 5 years old. The majority had the classical type of MSUD and dietary protein was restricted to between 0.6 gram/kg/day to 1 gram/kg/day of natural protein. Using the first pairs of observation for these 18 patients, the correlation coefficient was 0.76 (95% C1:0.462 to 0.907). This suggest a strong correlation between the two methods. It is recommended that further studies be done to determine the potential of the dried blood spot leu/ile level by TLC as an alternative method that can be used in the diagnosis and monitoring of MSUD patients especially in a developing country. (Author)

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

    International Nuclear Information System (INIS)

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

    2002-01-01

    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. (orig.)

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

    Science.gov (United States)

    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.

  20. Total reflection x-ray analysis of metals in blood samples

    International Nuclear Information System (INIS)

    Nakamura, Takuya; Matsui, Hiroshi; Kawamata, Masaya

    2009-01-01

    The sample preparation for TXRF (total reflection X-ray fluorescence) quantitative analysis of trace elements in human blood samples was investigated. In the TXRF analysis, a solution sample is dropped and dried on a flat substrate, and then the dried residue is measured. In this case, the dried residue should be flat not to disturb X-ray total reflection on the substrate. In addition, it is required to simply measure the whole blood sample by TXRF method, although a serum is analyzed in many cases. Thus, we studied the optimum conditions of the sample preparation of the whole blood by adding the pure water to apply Hemolysis phenomenon, where blood cells are destroyed due to different of the osmotic pressure, leading to flat residue. It was found that the best S/B ratio was obtained when the whole blood was diluted 8 times with pure water. Moreover, it was investigated the influence of the surface chemical condition of the glass substrate on the shape of the dried reside of the blood sample. When the surface of the glass substrate was hydrophilic, the shape of the dried residues was not uniform, as a result, the quantitative data of TXRF analysis gave a large deviation. On the other hand, when the surface of the glass was hydrophobic, the shape of the residue was almost uniform, as a result, a good reproducibility was obtained. Another problem was an outer ring of the dried residue of the blood. This uneven ring absorbs the primary X-rays, caused to low determined quantitative data. Thus, we tried the heating way of the dropped blood sample at a high temperature of 200 degrees. In this case, the blood sample was dried immediately, and a flat homogeneous dried residue was obtained without the outer ring. Using the optimized conditions for sample preparation, human blood sample was quantitatively measured by TXRF and ICP-AES. A good agreement was obtained in TXRF and ICP-AES determinations; however, the measurement of Cl and Br will be an advantage of TXRF, because

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

    Science.gov (United States)

    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.

  2. Utilizing Estimated Creatinine Excretion to Improve the Performance of Spot Urine Samples for the Determination of Proteinuria in Kidney Transplant Recipients.

    Directory of Open Access Journals (Sweden)

    Michael Ke Wang

    Full Text Available Agreement between spot and 24-hour urine protein measurements is poor in kidney transplant recipients. We investigated whether using formulae to estimate creatinine excretion rate (eCER, rather than assuming a standard creatinine excretion rate, would improve the estimation of proteinuria from spot urine samples in kidney transplant recipients.We measured 24 hour urine protein and albumin and spot albumin:creatinine (ACR and spot protein:creatinine (PCR in 181 Kidney transplant recipients." We utilized 6 different published formulae (Fotheringham, CKD-EPI, Cockcroft-Gault, Walser, Goldwasser and Rule to estimate eCER and from it calculated estimated albumin and protein excretion rate (eAER and ePER. Bias, precision and accuracy (within 15%, 30% and 50% of ACR, PCR, eAER, ePER were compared to 24-hour urine protein and albumin.ACR and PCR significantly underestimated 24-hour albumin and protein excretion (ACR Bias (IQR, -5.9 mg/day; p< 0.01; PCR Bias, (IQR, -35.2 mg/day; p<0.01. None of the formulae used to calculate eAER or ePER had a bias that was significantly different from the 24-hour collection (eAER and ePER bias: Fotheringham -0.3 and 7.2, CKD-EPI 0.3 and 13.5, Cockcroft-Gault -3.2 and -13.9, Walser -1.7 and 3.1, Goldwasser -1.3 and -0.5, Rule -0.6 and 4.2 mg/day respectively. The accuracy for ACR and PCR were lower (within 30% being 38% and 43% respectively than the corresponding values estimated by utilizing eCER (for eAER 46% to 49% and ePER 46-54%.Utilizing estimated creatinine excretion to calculate eAER and ePER improves the estimation of 24-hour albuminuria/proteinuria with spot urine samples in kidney transplant recipients.

  3. Single injection 51Cr EDTA plasma clearance determination in children using capillary blood samples

    International Nuclear Information System (INIS)

    Broechner-Mortensen, J.; Christoffersen, J.

    1977-01-01

    The reliability of a determination of the total 51 Cr EDTA plasma clearance (e) (and with it the glomerular filtration rate), by a simplified single injection method (injected dose: 4.5 μCi per kg b.w.) using capillary blood samples (0.2 ml), was investigated in twenty children. Clearance values determined from capillary blood samples did not differ significantly from those measured simultaneously from venous blood samples, the mean ratio+-SD being 1.02+-0.06(n = 10). The reproducibility (total day-to-day variation) of E determined from capillary blood samples was 6.7% in children with decreased renal function (n = 3) and 6.9% in children with normal renal function (n = 7). The present data indicate that the use of capillary blood samples is an accurate and very precise approach for determination of E in children. (Auth.)

  4. Relative Abundance of Proteins in Blood Plasma Samples from Patients with Chronic Cerebral Ischemia.

    Science.gov (United States)

    Kaysheva, Anna L; Kopylov, Artur T; Ponomarenko, Elena A; Kiseleva, Olga I; Teryaeva, Nadezhda B; Potapov, Alexander A; Izotov, Alexander А; Morozov, Sergei G; Kudryavtseva, Valeria Yu; Archakov, Alexander I

    2018-03-01

    A comparative protein profile analysis of 17 blood plasma samples from patients with ischemia and 20 samples from healthy volunteers was carried out using ultra-high resolution mass spectrometry. The analysis of measurements was performed using the proteomics search engine OMSSA. Normalized spectrum abundance factor (NSAF) in the biological samples was assessed using SearchGUI. The findings of mass spectrometry analysis of the protein composition of blood plasma samples demonstrate that the depleted samples are quite similar in protein composition and relative abundance of proteins. By comparing them with the control samples, we have found a small group of 44 proteins characteristic of the blood plasma samples from patients with chronic cerebral ischemia. These proteins contribute to the processes of homeostasis maintenance, including innate immune response unfolding, the response of a body to stress, and contribution to the blood clotting cascade.

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

    International Nuclear Information System (INIS)

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

    2005-01-01

    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

  6. The Use of Dried Blood Spots: A Potential Tool for the Introduction of a Neonatal Screening Program for Sickle Cell Anemia in Zambia.

    Science.gov (United States)

    Chindima, Nanjela; Nkhoma, Panji; Sinkala, Musalula; Zulu, Mildred; Kafita, Doris; Simakando, Marah; Mwaba, Florence; Mantina, Hamakwa; Mutale, Mubanga

    2018-01-01

    Sickle cell disease is a group of hemoglobin (Hb) disorders resulting from the inheritance of the sickle β-globin gene. It is the most common pathological Hb mutation worldwide with 75% being born in Sub-Saharan Africa. This study aims to determine if dried blood spots (DBSs) can be used for diagnosis of sickle cell in newborns. In Zambia, there is no neonatal screening program for sickle cell anemia (SCA), yet it has been proved that early diagnosis by newborn screening (NBS) using DBSs and access to comprehensive care results in survival to adulthood of over 96% of sickle cell patients. A cross-sectional study was carried out at the University Teaching Hospital to determine whether DBSs can be used to diagnose sickle cell using Hb electrophoresis. Results from DBSs stored for 2 weeks were then compared to those obtained using freshly collected whole blood. To evaluate performance characteristics, the following values were used: true positive, false positive, true negative, and false negative. Ninety-seven participants were included in this study. DBSs had a sensitivity of 100%, a specificity of 94.7%, positive predictive value of 96.7%, negative predictive value of 100%, overall efficiency of 97.9%, and a Kappa r 2 , P < 0.0001 in comparison to fresh whole blood which we used as the gold standard. The use of DBSs can be recommended for NBS of SCA in Zambia due to its high sensitivity, specificity, and stability of hemoglobin.

  7. Assessment of blood sample stability for complete blood count using the Sysmex XN-9000 and Mindray BC-6800 analyzers

    Directory of Open Access Journals (Sweden)

    Sabrina Buoro

    Full Text Available ABSTRACT BACKGROUND: Different hematological analyzers have different analytical performances that are often reflected in the criteria for sample stability of the complete blood count. This study aimed to assess the stability of several hematological parameters using the XN-9000 Sysmex and BC-6800 Mindray analyzers. METHODS: The impact of storage at room temperature and 4 °C was evaluated after 2, 4, 6, 8, 24, 36 and 48 h using ten normal and 40 abnormal blood samples. The variation from the baseline measurement was evaluated by the Steel-Dwass-Critchlow-Fligner test and by Bland-Altman plots, using quality specifications and critical difference as the total allowable variation. RESULTS: Red blood cells and reticulocyte parameters (i.e. hematocrit, mean corpuscular volume, mean corpuscular hemoglobin concentration, red blood cell distribution width, immature reticulocyte fractions, low-fluorescence reticulocytes, middle-fluorescence reticulocytes, high fluorescence mononuclear cells showed less stability compared to leukocyte and platelet parameters (except for monocyte count and mean platelet volume. The bias for hematocrit, mean corpuscular volume, mean corpuscular hemoglobin concentration and red blood cell distribution width coefficient of variation was higher than the critical difference after 8 h using both analyzers. CONCLUSION: Blood samples measured with both analyzers do not show analytically significant changes in up to 2 h of storage at room temperature and 4 °C. However, the maximum time for analysis can be extended for up to 8 h when the bias is compared to the critical difference.

  8. Simultaneous determination of 3-hydroxypropionic acid, methylmalonic acid and methylcitric acid in dried blood spots: Second-tier LC-MS/MS assay for newborn screening of propionic acidemia, methylmalonic acidemias and combined remethylation disorders.

    Directory of Open Access Journals (Sweden)

    Péter Monostori

    Full Text Available Increased propionylcarnitine levels in newborn screening are indicative for a group of potentially severe disorders including propionic acidemia (PA, methylmalonic acidemias and combined remethylation disorders (MMACBL. This alteration is relatively non-specific, resulting in the necessity of confirmation and differential diagnosis in subsequent tests. Thus, we aimed to develop a multiplex approach for concurrent determination of 3-hydroxypropionic acid, methylmalonic acid and methylcitric acid from the same dried blood spot (DBS as in primary screening (second-tier test. We also set out to validate the method using newborn and follow-up samples of patients with confirmed PA or MMACBL.The assay was developed using liquid chromatography-tandem mass spectrometry and clinically validated with retrospective analysis of DBS samples from PA or MMACBL patients.Reliable determination of all three analytes in DBSs was achieved following simple and fast (<20 min sample preparation without laborious derivatization or any additional pipetting steps. The method clearly distinguished the pathological and normal samples and differentiated between PA and MMACBL in all stored newborn specimens. Methylcitric acid was elevated in all PA samples; 3-hydroxypropionic acid was also high in most cases. Methylmalonic acid was increased in all MMACBL specimens; mostly together with methylcitric acid.A liquid chromatography-tandem mass spectrometry assay allowing simultaneous determination of the biomarkers 3-hydroxypropionic acid, methylmalonic acid and methylcitric acid in DBSs has been developed. The assay can use the same specimen as in primary screening (second-tier test which may reduce the need for repeated blood sampling. The presented preliminary findings suggest that this method can reliably differentiate patients with PA and MMACBL in newborn screening. The validated assay is being evaluated prospectively in a pilot project for extension of the German newborn

  9. Simultaneous determination of 3-hydroxypropionic acid, methylmalonic acid and methylcitric acid in dried blood spots: Second-tier LC-MS/MS assay for newborn screening of propionic acidemia, methylmalonic acidemias and combined remethylation disorders.

    Science.gov (United States)

    Monostori, Péter; Klinke, Glynis; Richter, Sylvia; Baráth, Ákos; Fingerhut, Ralph; Baumgartner, Matthias R; Kölker, Stefan; Hoffmann, Georg F; Gramer, Gwendolyn; Okun, Jürgen G

    2017-01-01

    Increased propionylcarnitine levels in newborn screening are indicative for a group of potentially severe disorders including propionic acidemia (PA), methylmalonic acidemias and combined remethylation disorders (MMACBL). This alteration is relatively non-specific, resulting in the necessity of confirmation and differential diagnosis in subsequent tests. Thus, we aimed to develop a multiplex approach for concurrent determination of 3-hydroxypropionic acid, methylmalonic acid and methylcitric acid from the same dried blood spot (DBS) as in primary screening (second-tier test). We also set out to validate the method using newborn and follow-up samples of patients with confirmed PA or MMACBL. The assay was developed using liquid chromatography-tandem mass spectrometry and clinically validated with retrospective analysis of DBS samples from PA or MMACBL patients. Reliable determination of all three analytes in DBSs was achieved following simple and fast (<20 min) sample preparation without laborious derivatization or any additional pipetting steps. The method clearly distinguished the pathological and normal samples and differentiated between PA and MMACBL in all stored newborn specimens. Methylcitric acid was elevated in all PA samples; 3-hydroxypropionic acid was also high in most cases. Methylmalonic acid was increased in all MMACBL specimens; mostly together with methylcitric acid. A liquid chromatography-tandem mass spectrometry assay allowing simultaneous determination of the biomarkers 3-hydroxypropionic acid, methylmalonic acid and methylcitric acid in DBSs has been developed. The assay can use the same specimen as in primary screening (second-tier test) which may reduce the need for repeated blood sampling. The presented preliminary findings suggest that this method can reliably differentiate patients with PA and MMACBL in newborn screening. The validated assay is being evaluated prospectively in a pilot project for extension of the German newborn screening

  10. Free thyroxine values in dried blood spots on filter paper in newborns are related to both gestational age and birth body weight.

    Science.gov (United States)

    Pacchiarotti, A; Bartalena, L; Chiovato, L; Falcone, M; Buratti, L; Ciampi, M; Giusti, L F; Grasso, L; Fenzi, G F; Martino, E

    1988-01-01

    The results of free thyroxine (FT4) measurements in dried blood spots on filter paper in 744 euthyroid newborns (616 at term, 128 preterm), 10 newborns with congenital hypothyroidism and 4 euthyroid newborns with congenital TBG deficiency are reported. FT4 was measured by column adsorption chromatography of free hormone followed by radioimmunoassay in the eluate. FT4 values averaged 24 +/- 0.2 pmol/L (mean +/- SE) in euthyroid newborns, 23.0 +/- 0.9 pmol/L in euthyroid newborns with TBG deficiency (p = NS), and 5.7 +/- 0.4 pmol/L in hypothyroid newborns (p less than 0.001 vs both groups). Total T4 (TT4) values in newborns with TBG deficiency were not different from those in hypothyroid newborns, but were significantly lower than those in euthyroid newborns without TBG abnormalities. FT4 values were higher in full-term newborns than in preterm newborns (25.2 +/- 0.3 vs 21.2 +/- 0.5 pmol/L, p less than 0.001). In both full-term and preterm newborns FT4 values in dried blood spots increased with birth body weight (bbw), virtually plateauing when bbw was greater than 2,500 g. The cut-off values established on the basis of the bbw (8.0 and 13.1 pmol/L for a bbw of less than or equal to 2,500 g and greater than 2,500 g, respectively) showed higher specificity and predictive value of positive results than the cut-off values based on the gestational age. In any case, the sensitivity, specificity and predictive values of FT4 determinations proved to be higher than those of TT4 and TSH measurements.(ABSTRACT TRUNCATED AT 250 WORDS)

  11. [Detection of Candida DNA in simulated blood samples by polymerase chain reaction].

    Science.gov (United States)

    Ozer, Sinem; Yücesoy, Mine

    2007-07-01

    Although blood culture method is accepted as gold standard in the laboratory diagnosis of invasive candidiasis seen in immunocompromised patients, cultivation of blood is considered as not a reliable and rapid method for the diagnosis of candidemia, since it may be negative in approximately half of the patients, slow growth rate of Candida in routine culture media and requirement of large amounts of blood for the isolation. The aim of this study was to detect Candida DNA in simulated blood samples by using polymerase chain reaction (PCR). Simulated samples were prepared by using blood samples of healthy volunteers. These samples were inoculated into tubes with EDTA and BACTEC 9240 blood culture bottles in which no growth was detected and with standard strains of C. albicans, C. tropicalis, C. parapsilosis, C. krusei, Escherichia coli and Staphylococcus aureus together with the clinical isolates of Aspergillus fumigatus, C. kefyr, C. glabrata, C. lusitaniae, C. guilliermondii and Rhodotorula sp. Additionally, blood culture samples of 23 cases whose blood culture bottles signaled as positive and revealed growth of Candida in agar plates were examined. DNA extraction of all samples were performed according to the standard procedure proposed by the MN Nucleospin Tissue Kit (Macherey-Nagel, Germany) for tissue samples; following the pre-treatment with erythrocyte, leukocyte and fungus cell wall lysis buffers. DNAs were amplified with PCR, using primers specific for the 5S rDNA region (PCon 1 and PCon 2 primers) and PCR products were obtained by electrophoresis in 2% agarose gel. Presence of a 105 base pair (bp) product was considered as positive. The lowest detection limit of PCR has been determined as 10(2)-10(3) cfu/ml Candida for our simulated samples. The presence of a 105 bp band has been observed in samples prepared with all Candida strains included in the study. Blood samples spiked with E. coli, S. aureus, A. fumigatus and Rhodotorula sp. and negative blood

  12. Microcapillary blood sampling for serological examinations by radioimmunoassay (RIA) and enzyme immunoassay (ELISA)

    Energy Technology Data Exchange (ETDEWEB)

    Rodak, L.; Smid, B.; Valicek, L.; Jurak, E. (Vyzkumny Ustav Veterinarniho Lekarstvi, Brno-Medlanky (Czechoslovakia))

    1984-01-01

    Methods were tested of sampling blood and blood serum for serological examinations on filtration paper and into heparinized glass capillaries with transfer into the dilution solution of the given composition. Samples were also examined for ACH virus antibodies. The suitability of the sampling was verified by an examination of samples using ELISA and RIA methods. The results showed the suitability of sampling using microcapillaries. The titres of virus antibodies found using the ELISA and RIA methods were identical and the sensitivity of antibody detection was not reduced even after the sample had been stored for 60 days at a temperature of 20 degC.

  13. Microcapillary blood sampling for serological examinations by radioimmunoassay (RIA) and enzyme immunoassay (ELISA)

    International Nuclear Information System (INIS)

    Rodak, L.; Smid, B.; Valicek, L.; Jurak, E.

    1984-01-01

    Methods were tested of sampling blood and blood serum for serological examinations on filtration paper and into heparinized glass capillaries with transfer into the dilution solution of the given composition. Samples were also examined for ACH virus antibodies. The suitability of the sampling was verified by an examination of samples usiOg ELISA and RIA methods. The results showed the suitability of sampling using microcapillaries. The titres of virus antibodies found using the ELISA and RIA methods were identical and the sensitivity of antibody detection was not reduced even after the sample had been stored for 60 days at a temperature of 20 degC. (B.S.)

  14. [Responsibility of the physician for blood samples according to paragraph 81 a StPO?].

    Science.gov (United States)

    Blank, J H

    1992-03-01

    On principle no physician is under obligation to take blood-tests prescribed in Art. 81 a StPO (Code of Criminal Procedure). However, this principle is not valid without exception. Hence to be appointed expert by a judge or a public prosecutor shall give the reason for a legal pledge. An indirect obligation--but not to police authorities--may result from a contractually stipulated assumption or from being part of the Public Health Service staff. For the physician being employed in a hospital, the special rule 2 c fig. 3 par. 1 BAT (Federal Employees Tariff Treaty) is valid, which stipulates an obligation to take blood-tests, even if a contractual agreement is missing. Violation of contract and offence against legal pledges may justify giving notice to the physician. However, the obligation of taking blood-tests is not unlimited. Within the pale of law rights of refusal are stated in Art. 22, 24, 52, 72, 74 StPO (Code of Criminal Procedure). Moreover a refusal is justified, if the manoeuvre is injurious to health of the accused under consideration. In this case the physician on the spot taking blood-tests has got the exclusive responsibility, which cannot be shifted on to other authorities. In case of personally justified refusal the physician does not have to expect any prejudices. A physician refusing to take blood-tests does not incur a penalty under Art. 258 or Art. 258 a StGB (Criminal Code).

  15. Use of Dried Blood Spots for Estimating Children?s Exposures to Heavy Metals in Epidemiological Research

    Science.gov (United States)

    Background: Children’s exposures to arsenic (As), lead (Pb), mercury (Hg), and cadmium (Cd) are of particular concern in early-life. Exposures to heavy metals are traditionally measured in whole venous blood, which is costly and invasive. As an alternative we describe a met...

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

    Science.gov (United States)

    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. Copyright © 2014. Published by Elsevier B.V.

  17. A method to quantitate cerebral blood flow using a rotating gamma camera and iodine-123 iodoamphetamine with one blood sampling

    International Nuclear Information System (INIS)

    Iida, Hidehiro; Itoh, Hiroshi; Bloomfield, P.M.; Munaka, Masahiro; Higano, Shuichi; Murakami, Matsutaro; Inugami, Atsushi; Eberl, S.; Aizawa, Yasuo; Kanno, Iwao; Uemura, Kazuo

    1994-01-01

    A method has been developed to quantitate regional cerebral blood blow (rCBF) using iodine-123-labelled N-isopropyl-p-iodoamphetamine (IMP). This technique requires only two single-photon emission tomography (SPET) scans and one blood sample. Based on a two-compartment model, radioactivity concentrations in the brain for each scan time are calculated. A standard input function has been generated by combining the input functions from 12 independent studies prior to this work to avoid frequent arterial blood sampling, and one blood sample is taken at 10 min following IMP administration for calibration of the standard arterial input function. This calibration time was determined such that the integration of the first 40 min of the calibrated, combined input function agreed best with those from 12 individual input functions (the difference was 5.3% on average). This method was applied to eight subjects (two normals and six patients with cerebral infarction), and yielded rCBF values which agreed well with those obtained by a positron emission tomography H 2 15 O autoradiography method. This method was also found to provide rCBF values that were consistent with those obtained by the non-linear least squares fitting technique and those obtained by conventional microsphere model analysis. The optimum SPET scan times were found to be 40 and 180 min for the early and delayed scans, respectively. These scan times allow the use of a conventional rotating gamma camera for clinical purposes. V d values ranged between 10 and 40 ml/g depending on the pathological condition, thereby suggesting the importance of measuring V d for each ROI. In conclusion, optimization of the blood sampling time and the scanning time enabled quantitative measurement of rCBF with two SPET scans and one blood sample. (orig.)

  18. Detection of drugs in 275 alcohol-positive blood samples of Korean drivers.

    Science.gov (United States)

    Kim, Eunmi; Choe, Sanggil; Lee, Juseon; Jang, Moonhee; Choi, Hyeyoung; Chung, Heesun

    2016-08-01

    Since driving under the influence of drugs (DUID) is as dangerous as drink-driving, many countries regulate DUID by law. However, laws against the use of drugs while driving are not yet established in Korea. In order to investigate the type and frequency of drugs used by drivers in Korea, we analyzed controlled and non-controlled drugs in alcohol-positive blood samples. Total 275 blood samples were taken from Korean drivers, which were positive in roadside alcohol testing. The following analyses were performed: blood alcohol concentrations by GC; screening for controlled drugs by immunoassay and confirmation for positive samples by GC-MS. For the detection of DUID related drugs in blood samples, a total of 49 drugs were selected and were examined by GC-MS. For a rapid detection of these drugs, an automated identification software called "DrugMan" was used. Concentrations of alcohol in 275 blood samples ranged from 0.011 to 0.249% (average 0.119%). Six specimens showed positive results by immunoassay: one methamphetamine and five benzodiazepines I. By GC-MS confirmation, only benzodiazepines in four cases were identified, while methamphetamine and benzodiazepine in two cases were not detected from the presumptive positive blood samples. Using DrugMan, four drugs were detected; chlorpheniramine (5)*, diazepam (4), dextromethorphan (1) and doxylamine (1). In addition, ibuprofen (1), lidocaine (1) and topiramate (1) were also detected as general drugs in blood samples ('*' indicates frequency). The frequency of drug abuse by Korean drivers was relatively low and a total 14 cases were positive in 275 blood samples with a ratio of 5%. However it is necessary to analyze more samples including alcohol negative blood, and to expand the range of drug lists to get the detailed information. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  19. Evaluation of the use of purine derivatives:creatinine ratio in spot urine samples as an index of microbial protein supply in Yerli Kara crossbred cattle

    International Nuclear Information System (INIS)

    Cetinkaya, N.; Ozdemir, H.; Gucus, A.I.; Ozcan, H.; Sogut, A.; Yaman, S.

    2004-01-01

    In Experiment I, response of daily purine derivatives (PD) excretion to feed intake in Yerli Kara crossbred (YK-C) cattle on state farms was measured. Animals were fed a mixed diet containing 30% wheat straw and 70% compounded feed. Crude protein and organic matter contents of the diet were 12.4% and 95%, respectively. In Experiment II, spot urine sampling technique was applied at state farm. Four Yerli Kara crossbred bulls with a mean live weight of 211.0 ± 41.3 kg were used. The experimental design, feeding and diet were the same as in Experiment I. The treatments were allocated according to a 4 x 4 Latin Square design. In Experiment III, spot urine sampling technique was applied at smallholder farms. Two to three kg of compound feed (crude protein 12%) containing 65% barley, 25% bran, 6% sunflower seed meal, 3% marmer dust and 1% mineral and vitamin mixture was offered in two parts, one in the morning (0730 h) and the other in the afternoon (1700 h). The ingredients in the compound feed were similar for all animals, but animals in Groups I, II and III received 1 to 2 kg/d of straw (crude protein 3%), grass hay (crude protein 7%), or both straw and grass hay respectively. In Experiment I, a significant correlation (R 2 =0.99) between PD excretion (Y, mmol/d) and digestible organic matter intake, DOMI (X, kg/d) for YK-C cattle was observed (Y = 12.5 + 19.7 X). Moreover, daily PD excretion (mmol/d) was correlated with the PDC index, which was defined as [PD molar concentration] / [Creatinine molar concentration] x kgW 0.75 . In Experiment II, the PDC index increased with level of intake. The coefficient of variation due to time of sampling for uric acid, allantoin, PD, creatinine, total-N, the PDC Index in spot urine samples were less than 5%. In Experiment III, the PDC index were 49.95 ± 13.5, 45.6 ± 13.0, 48.95 ± 15.3 for the three groups respectively. These values were similar to those for 60% intake level in Experiment I. Using the equation DOMI = 344 + 48

  20. [Evaluation of the test strip Reflotron ALP (alkaline phosphatase) for blood samples of dogs and cats].

    Science.gov (United States)

    Hirschberger, J; Dietz, J; Baumeister, C; Kraft, M

    1998-09-01

    The test strip Reflotron ALP (alcaline phosphatase) was evaluated for use in canine and feline heparinized blood samples. The within-run and the day-to-day precision of blood and plasma samples was excellent. A haematocrit up to 60% of canine blood and up to 50% of feline blood had no influence on the measurements. Reflotron ALP was compared with ALP on Hitachi 717 (Boehringer Mannheim, Mannheim, Germany). The correlation of both methods was good for canine and feline blood samples. Reflotron ALP was higher than Hitachi ALP in both species. Despite an enormous deviation between Reflotron ALP and the reference method of canine and feline blood samples within the reference range, Reflotron ALP is a suited test for the detection of elevated ALP activity in canine and feline blood samples. Differences of the ALP activity might be caused by ALP isoenzymes. The activity of ALP isoenzymes depends on the method. The buffer systems of Reflotron ALP and Hitachi ALP are different. Significant exceeding of the reference range was reliably detected. In this investigation the study results for heparinized whole blood, heparinized plasma and serum are approximately the same.

  1. Effect of concentrate supplementation on feed consumption, nutrient utilization and blood metabolite profile in captive spotted deer (Axis axis) fed oat (Avena sativa) and berseem (Trifolium alexandrinum) fodders based diet.

    Science.gov (United States)

    Suresh, C; Das, A; Katole, Shrikant; Saini, Mohini; Swarup, D

    2013-03-01

    This experiment was conducted to determine the optimum level of a maize-soybean meal-wheat bran concentrate supplement fed to captive spotted deer fed an oat and berseem fodder-based diet. Twelve adult spotted deer [64-76 kg body weight (BW)] were distributed into three groups of four each and were housed individually. A diet consisting of 5 kg of oat fodder and 5.5 kg of berseem fodder was offered to each one of the experimental animals. The animal in group I received no supplementary concentrate, whereas, those in groups II and III received 0.5 and 1 kg of supplementary concentrate, respectively. A 60 days digestibility trial was conducted with a 5 days collection period on Days 55-59 of the trial. Blood samples were collected from all animals on Day 60 of the experiment. Average daily dry matter intake (DMI) was 1,224, 1,613, and 1,574 g/day in groups I, II, and III, respectively. Dry matter (DM) and organic matter (OM) intake was lowest (P II and I. Digestibility of neutral detergent fiber was highest (P II. Digestibility of OM and CP was lowest (P energy was highest (P II (69.3%) and I (66.2%). Digestible energy (DE) intake (kcal/kg BW(0.75) ) was highest (P II (180.9) and I (129.8). Initial BW was 72.7, 72.5, and 71.0 kg, whereas, final BW was 71.0, 72.7, and 73.5 kg, in groups I, II and III, respectively. Average daily change in body mass was significantly (P II, and III, respectively. Blood glucose and cholesterol concentration was highest (P II and I. Serum concentration of Cu and Zn was highest (P II and I. Supplementation of forage only diet with 0.5 kg of concentrate mixture increased intake and digestibility of nutrients, without change in body mass. Animals fed 1 kg of supplementary concentrate received energy in excess of requirements, were consistently gaining body mass and were prone to obesity. Thus, it is a right strategy to supplement forage only diet of captive spotted deer with 0.5 kg of concentrate. © 2013

  2. Human blood RNA stabilization in samples collected and transported for a large biobank

    Directory of Open Access Journals (Sweden)

    Duale Nur

    2012-09-01

    Full Text Available Abstract Background The Norwegian Mother and Child Cohort Study (MoBa is a nation-wide population-based pregnancy cohort initiated in 1999, comprising more than 108.000 pregnancies recruited between 1999 and 2008. In this study we evaluated the feasibility of integrating RNA analyses into existing MoBa protocols. We compared two different blood RNA collection tube systems – the PAXgene™ Blood RNA system and the Tempus™ Blood RNA system - and assessed the effects of suboptimal blood volumes in collection tubes and of transportation of blood samples by standard mail. Endpoints to characterize the samples were RNA quality and yield, and the RNA transcript stability of selected genes. Findings High-quality RNA could be extracted from blood samples stabilized with both PAXgene and Tempus tubes. The RNA yields obtained from the blood samples collected in Tempus tubes were consistently higher than from PAXgene tubes. Higher RNA yields were obtained from cord blood (3 – 4 times compared to adult blood with both types of tubes. Transportation of samples by standard mail had moderate effects on RNA quality and RNA transcript stability; the overall RNA quality of the transported samples was high. Some unexplained changes in gene expression were noted, which seemed to correlate with suboptimal blood volumes collected in the tubes. Temperature variations during transportation may also be of some importance. Conclusions Our results strongly suggest that special collection tubes are necessary for RNA stabilization and they should be used for establishing new biobanks. We also show that the 50,000 samples collected in the MoBa biobank provide RNA of high quality and in sufficient amounts to allow gene expression analyses for studying the association of disease with altered patterns of gene expression.

  3. Human blood RNA stabilization in samples collected and transported for a large biobank

    Science.gov (United States)

    2012-01-01

    Background The Norwegian Mother and Child Cohort Study (MoBa) is a nation-wide population-based pregnancy cohort initiated in 1999, comprising more than 108.000 pregnancies recruited between 1999 and 2008. In this study we evaluated the feasibility of integrating RNA analyses into existing MoBa protocols. We compared two different blood RNA collection tube systems – the PAXgene™ Blood RNA system and the Tempus™ Blood RNA system - and assessed the effects of suboptimal blood volumes in collection tubes and of transportation of blood samples by standard mail. Endpoints to characterize the samples were RNA quality and yield, and the RNA transcript stability of selected genes. Findings High-quality RNA could be extracted from blood samples stabilized with both PAXgene and Tempus tubes. The RNA yields obtained from the blood samples collected in Tempus tubes were consistently higher than from PAXgene tubes. Higher RNA yields were obtained from cord blood (3 – 4 times) compared to adult blood with both types of tubes. Transportation of samples by standard mail had moderate effects on RNA quality and RNA transcript stability; the overall RNA quality of the transported samples was high. Some unexplained changes in gene expression were noted, which seemed to correlate with suboptimal blood volumes collected in the tubes. Temperature variations during transportation may also be of some importance. Conclusions Our results strongly suggest that special collection tubes are necessary for RNA stabilization and they should be used for establishing new biobanks. We also show that the 50,000 samples collected in the MoBa biobank provide RNA of high quality and in sufficient amounts to allow gene expression analyses for studying the association of disease with altered patterns of gene expression. PMID:22988904

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

    Science.gov (United States)

    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.

  5. Serum, plasma, and dried blood spot high sensitivity C-reactive protein enzyme immunoassay for population research

    OpenAIRE

    Brindle, Eleanor; Fujita, Masako; Shofer, Jane; O’Connor, Kathleen A.

    2010-01-01

    C-reactive protein (CRP) is used as a biomarker of morbidity and mortality risk in studies of population health, and is essential to interpretation of several micronutrient biomarkers. There is thus need for a robust high sensitivity CRP (hsCRP) measurement method for large-scale, non-clinical studies. We developed an efficient, inexpensive assay suitable for quantifying CRP across the physiological range using any blood specimen type. The ELISA uses readily available monoclonal antibodies to...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. Copyright © 2014 Elsevier Inc. All rights reserved.

  8. Age Spots

    Science.gov (United States)

    ... for Every Season How to Choose the Best Skin Care Products In This Section Dermatologic Surgery What is dermatologic ... for Every Season How to Choose the Best Skin Care Products Age Spots Treatment Options Learn more about treatment ...

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

    Science.gov (United States)

    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.

  10. Chlamydia trachomatis antibody detection in home-collected blood samples for use in epidemiological studies.

    NARCIS (Netherlands)

    Hoenderboom, B M; van Ess, E F; van den Broek, I V F; van Loo, I H M; Hoebe, C J P A; Ouburg, S; Morré, S A

    Capillary blood collected in serum tubes was subjected to centrifugation delay while stored at room temperature. Chlamydia trachomatis (CT) IgG concentrations in aliquoted serum of these blood samples remained stable for seven days after collection. CT IgG concentrations can reliably be measured in

  11. Leukocyte count affects expression of reference genes in canine whole blood samples

    NARCIS (Netherlands)

    Piek, C.J.; Brinkhof, B.; Rothuizen, J.; Dekker, A.; Penning, L.C.

    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

  12. Evaluation of some toxic metals in blood samples of smokers in ...

    African Journals Online (AJOL)

    Purpose: To determine some toxic elements in the blood of cigarette and tobacco pipe smokers in Riyadh, Saudi Arabia. Methods: The study setting was Riyadh, the capital city of Saudi Arabia, Riyadh City. Male volunteers, aged 20 - 58 year, whose blood samples were collected, were classified into three groups of ...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  15. An LC–MS/MS-Based Method for the Quantification of Pyridox(am)ine 5′-Phosphate Oxidase Activity in Dried Blood Spots from Patients with Epilepsy

    Science.gov (United States)

    2017-01-01

    We report the development of a rapid, simple, and robust LC–MS/MS-based enzyme assay using dried blood spots (DBS) for the diagnosis of pyridox(am)ine 5′-phosphate oxidase (PNPO) deficiency (OMIM 610090). PNPO deficiency leads to potentially fatal early infantile epileptic encephalopathy, severe developmental delay, and other features of neurological dysfunction. However, upon prompt treatment with high doses of vitamin B6, affected patients can have a normal developmental outcome. Prognosis of these patients is therefore reliant upon a rapid diagnosis. PNPO activity was quantified by measuring pyridoxal 5′-phosphate (PLP) concentrations in a DBS before and after a 30 min incubation with pyridoxine 5′-phosphate (PNP). Samples from 18 PNPO deficient patients (1 day–25 years), 13 children with other seizure disorders receiving B6 supplementation (1 month–16 years), and 37 child hospital controls (5 days–15 years) were analyzed. DBS from the PNPO-deficient samples showed enzyme activity levels lower than all samples from these two other groups as well as seven adult controls; no false positives or negatives were identified. The method was fully validated and is suitable for translation into the clinical diagnostic arena. PMID:28782931

  16. Effects of Transport and Storage Conditions on Gene Expression in Blood Samples.

    Science.gov (United States)

    Malentacchi, Francesca; Pizzamiglio, Sara; Wyrich, Ralf; Verderio, Paolo; Ciniselli, Chiara; Pazzagli, Mario; Gelmini, Stefania

    2016-04-01

    Inappropriate handling of blood samples might induce or repress gene expression and/or lead to RNA degradation affecting downstream analysis. In particular, sample transport is a critical step for biobanking or multicenter studies because of uncontrolled variables (i.e., unstable temperature). We report the results of a pilot study implemented within the EC funded SPIDIA project, aimed to investigate the role of transport and storage of blood samples containing and not containing an RNA stabilizer. Blood was collected from a single donor both in EDTA and in PAXgene Blood RNA tubes. Half of the samples were sent to a second laboratory both at room temperature and at 4°C, whereas the remaining samples were stored at room temperature and at 4°C. Gene expression of selected genes (c-FOS, IL-1β, IL-8, and GAPDH) known to be induced or repressed by ex vivo blood handling and of blood-mRNA quality biomarkers identified and validated within the SPIDIA project, which allow for monitoring changes in unstabilized blood samples after collection and during transport and storage, were analyzed by RT-qPCR. If the shipment of blood in tubes not containing RNA stabilizer is not performed under a stable condition, gene profile studies can be affected by the effects of transport. Moreover, also controlled temperature shipment (4°C) can influence the expression of specific genes if blood is collected in tubes not containing a stabilizer. The use of dedicated biomarkers or time course experiments should be performed in order to verify potential bias on gene expression analysis due to sample shipment and storage conditions. Alternatively, the use of RNA stabilizer containing tubes can represent a reliable option to avoid ex vivo RNA changes.

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

    Science.gov (United States)

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

    2017-12-01

    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.

  18. Na2EDTA anticoagulant impaired blood samples from the teleost Piaractus mesopotamicus

    Directory of Open Access Journals (Sweden)

    Thaís Heloisa Vaz Farias

    2016-05-01

    Full Text Available Abstract: The present study aimed to evaluate the effects of Na heparin and Na2EDTA on blood of Piaractus mesopotamicus (360.7±42.4g, 26.4±1.0cm. Twenty fishes were sampled in two experiment trials, ten for erythrocyte fragility analysis and ten for hematologic and plasma biochemical study. The blood collected by venous-caudal puncture was fractioned and stored in anticoagulants solution: Na2EDTA 10%, Na2EDTA 3%, Na heparin 5000 IU and Na heparin 100 IU. Plasmatic levels of calcium presented in the Na2EDTA stored samples were about 80% lower than both heparin groups. Blood samples of P. mesopotamicus stored with Na2EDTA demonstrated increase in the hematocrit and MCV, and decrease in MCHC. The dose-response effect was observed in this study. The results are reinforced by the higher levels of plasmatic protein and hemolysis presented in the Na2EDTA 10% stored blood, confirming the deleterious effect of this anticoagulant treatment on the quality of blood samples. Na2EDTA is not indicated to store P. mesopotamicus blood samples, but sodium heparin at 100 IU is the most recommended anticoagulant, since this treatment presented the lower rate of alterations in the stored blood.

  19. A duplex PCR for rapid and simultaneous detection of Brucella spp. in human blood samples.

    Science.gov (United States)

    Mirnejad, Reza; Mohamadi, Mozafar; Piranfar, Vahbeh; Mortazavi, Seied Mojtaba; Kachuei, Reza

    2013-06-01

    To design a duplex PCR for rapid and simultaneous detection of Brucella species. in human blood samples. Fifty-two peripheral bloods samples were collected from suspicious patients with brucellosis. Following DNA extraction, PCR assay were performed, using three primers that could simultaneously identify and differentiate three major species of pathogenic Brucella in humans and animals. Of the 52 peripheral bloods samples tested, 25 sample (48%) showed positive reactions in PCR. Twelve samples were positive for Brucella abortus 39 (B. abortus 39) (23%), 13 for Brucella melitensis 39 (B. melitensis 39) (25%) and 0 for Brucella ovis 39 (B. ovis 39) (0%). This work demonstrates that in case where specific primers were utilized, duplex PCR has proved to be a simple, fast, and relatively inexpensive method for simultaneous detection of important species of Brucella in clinical samples. Copyright © 2013 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

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

    DEFF Research Database (Denmark)

    Bach, Cathrine Carlsen; Henriksen, Tine Brink; Bossi, Rossana

    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...

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

    OpenAIRE

    Lenicek Krleza, Jasna

    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 t...

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

    Science.gov (United States)

    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% CaCl 2 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.

  3. Hot-spot detection and calibration of a scanning thermal probe with a noise thermometry gold wire sample

    NARCIS (Netherlands)

    Gaitas, A.; Wolgast, S.; Covington, E.; Kurdak, C.

    2013-01-01

    Measuring the temperature profile of a nanoscale sample using scanning thermal microscopy is challenging due to a scanning probe's non-uniform heating. In order to address this challenge, we have developed a calibration sample consisting of a 1-?m wide gold wire, which can be heated electrically by

  4. Survivability of Existing Peripheral Intravenous Access Following Blood Sampling in a Pediatric Population.

    Science.gov (United States)

    O'Neil, Sheree W; Friesen, Mary Ann; Stanger, Debra; Trickey, Amber Williams

    2018-03-07

    Although pediatric patients report venipuncture as their most feared experience during hospitalization, blood sampling from peripheral intravenous accesses (PIVs) is not standard of care. Blood sampling from PIVs has long been considered by healthcare personnel to harm the access. In an effort to minimize painful procedures, pediatric nursing staff conducted a prospective, observational study to determine if blood sampling using existing PIVs resulted in the loss of the access. The ability to obtain the sample from the PIV was measured along with patient and PIV characteristics. Specimen collection using 100 existing PIVs was attempted on pediatric inpatients. Each PIV was observed for functionality, infiltration, occlusion, and dislodgement following collection and again in 4h. Frequencies of PIV loss and successful blood sampling were calculated. Patient age, PIV gauge, access site, and PIV age were evaluated for associations with successful sampling using chi-square tests, Fisher's exact tests, and logistic regression. PIV survivability was reported at 99%. The ability to obtain a complete specimen was reported at 76% and found to be significantly related to PIV age and site. Size of PIV and patient's age were not significantly related to successful sampling. Encouraging rates of PIV survivability and collectability suggest blood sampling from PIVs to be a valuable technique to minimize painful and distressful procedures. Nursing practice was changed in this pediatric department. Patients and families are saved the pain and distress of venipuncture. Nurses reported saving time and personal distress by avoiding the venipuncture procedure. Copyright © 2018 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    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.

  6. Dried blood spot analysis for therapeutic drug monitoring of linezolid in patients with multidrug-resistant tuberculosis

    NARCIS (Netherlands)

    Vu, D H; Bolhuis, M S; Koster, R A; Greijdanus, B; de Lange, W C M; van Altena, R; Brouwers, J R B J; Uges, D R A; Alffenaar, J W C

    2012-01-01

    Linezolid is a promising antimicrobial agent for the treatment of multidrug-resistant tuberculosis (MDR-TB), but its use is limited by toxicity. Therapeutic drug monitoring (TDM) may help to minimize toxicity while adequate drug exposure is maintained. Conventional plasma sampling and monitoring

  7. Stability of heparin blood samples during transport based on defined pre-analytical quality goals

    DEFF Research Database (Denmark)

    Jensen, Esther A; Stahl, Marta; Brandslund, Ivan

    2008-01-01

    two periods (winter and summer). Transport conditions (mail, courier pick-up, or brought to hospital by public coach), storage time, storage temperature and centrifugation requirements were different in the two counties. Results were tested for deviation from a "0-sample", the blood sample taken......, centrifuged and separated at the doctor's office within 45-60 min. This sample was considered as the best estimate of a comparison value. RESULTS: The pre-set quality goals were fulfilled for all the investigated components for samples transported to hospital by courier either as whole blood or as "on gel...... whole blood if the above mentioned conditions are met. There is no need for centrifugation in the primary sector. Neither mailing of samples with plasma "on gel" nor public transport by coach bus fulfil our analytical goals....

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

    Science.gov (United States)

    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 Psprocedures except for JV-XKA and JV-MMF. Plasma NEFAs increased to 52% (Pprocedure 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 Psprocedures 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 the design and interpretation of blood sampling experiments in laboratory animals. (c) 2010. Published by Elsevier Inc.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  11. A simple method for measurement of cerebral blood flow using 123I-IMP SPECT with calibrated standard input function by one point blood sampling. Validation of calibration by one point venous blood sampling as a substitute for arterial blood sampling

    International Nuclear Information System (INIS)

    Ito, Hiroshi; Akaizawa, Takashi; Goto, Ryoui

    1994-01-01

    In a simplified method for measurement of cerebral blood flow using one 123 I-IMP SPECT scan and one point arterial blood sampling (Autoradiography method), input function is obtained by calibrating a standard input function by one point arterial blood sampling. A purpose of this study is validation of calibration by one point venous blood sampling as a substitute for one point arterial blood sampling. After intravenous infusion of 123 I-IMP, frequent arterial and venous blood sampling were simultaneously performed on 12 patients of CNS disease without any heart and lung disease and 5 normal volunteers. The radioactivity ratio of venous whole blood which obtained from cutaneous cubital vein to arterial whole blood were 0.76±0.08, 0.80±0.05, 0.81±0.06, 0.83±0.11 at 10, 20, 30, 50 min after 123 I-IMP infusion, respectively. The venous blood radioactivities were always 20% lower than those of arterial blood radioactivity during 50 min. However, the ratio which obtained from cutaneous dorsal hand vein to artery were 0.93±0.02, 0.94±0.05, 0.98±0.04, 0.98±0.03, at 10, 20, 30, 50 min after 123 I-IMP infusion, respectively. The venous blood radioactivity was consistent with artery. These indicate that arterio-venous difference of radioactivity in a peripheral cutaneous vein like a dorsal hand vein is minimal due to arteriovenous shunt in palm. Therefore, a substitution by blood sampling from cutaneous dorsal hand vein for artery will be possible. Optimized time for venous blood sampling evaluated by error analysis was 20 min after 123 I-IMP infusion, which is 10 min later than that of arterial blood sampling. (author)

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

    Science.gov (United States)

    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.

  13. Intraosseous blood samples for point-of-care analysis: agreement between intraosseous and arterial analyses.

    Science.gov (United States)

    Jousi, Milla; Saikko, Simo; Nurmi, Jouni

    2017-09-11

    Point-of-care (POC) testing is highly useful when treating critically ill patients. In case of difficult vascular access, the intraosseous (IO) route is commonly used, and blood is aspirated to confirm the correct position of the IO-needle. Thus, IO blood samples could be easily accessed for POC analyses in emergency situations. The aim of this study was to determine whether IO values agree sufficiently with arterial values to be used for clinical decision making. Two samples of IO blood were drawn from 31 healthy volunteers and compared with arterial samples. The samples were analysed for sodium, potassium, ionized calcium, glucose, haemoglobin, haematocrit, pH, blood gases, base excess, bicarbonate, and lactate using the i-STAT® POC device. Agreement and reliability were estimated by using the Bland-Altman method and intraclass correlation coefficient calculations. Good agreement was evident between the IO and arterial samples for pH, glucose, and lactate. Potassium levels were clearly higher in the IO samples than those from arterial blood. Base excess and bicarbonate were slightly higher, and sodium and ionised calcium values were slightly lower, in the IO samples compared with the arterial values. The blood gases in the IO samples were between arterial and venous values. Haemoglobin and haematocrit showed remarkable variation in agreement. POC diagnostics of IO blood can be a useful tool to guide treatment in critical emergency care. Seeking out the reversible causes of cardiac arrest or assessing the severity of shock are examples of situations in which obtaining vascular access and blood samples can be difficult, though information about the electrolytes, acid-base balance, and lactate could guide clinical decision making. The analysis of IO samples should though be limited to situations in which no other option is available, and the results should be interpreted with caution, because there is not yet enough scientific evidence regarding the agreement of IO

  14. SPOT Program

    Science.gov (United States)

    Smith, Jason T.; Welsh, Sam J.; Farinetti, Antonio L.; Wegner, Tim; Blakeslee, James; Deboeck, Toni F.; Dyer, Daniel; Corley, Bryan M.; Ollivierre, Jarmaine; Kramer, Leonard; hide

    2010-01-01

    A Spacecraft Position Optimal Tracking (SPOT) program was developed to process Global Positioning System (GPS) data, sent via telemetry from a spacecraft, to generate accurate navigation estimates of the vehicle position and velocity (state vector) using a Kalman filter. This program uses the GPS onboard receiver measurements to sequentially calculate the vehicle state vectors and provide this information to ground flight controllers. It is the first real-time ground-based shuttle navigation application using onboard sensors. The program is compact, portable, self-contained, and can run on a variety of UNIX or Linux computers. The program has a modular objec-toriented design that supports application-specific plugins such as data corruption remediation pre-processing and remote graphics display. The Kalman filter is extensible to additional sensor types or force models. The Kalman filter design is also strong against data dropouts because it uses physical models from state and covariance propagation in the absence of data. The design of this program separates the functionalities of SPOT into six different executable processes. This allows for the individual processes to be connected in an a la carte manner, making the feature set and executable complexity of SPOT adaptable to the needs of the user. Also, these processes need not be executed on the same workstation. This allows for communications between SPOT processes executing on the same Local Area Network (LAN). Thus, SPOT can be executed in a distributed sense with the capability for a team of flight controllers to efficiently share the same trajectory information currently being computed by the program. SPOT is used in the Mission Control Center (MCC) for Space Shuttle Program (SSP) and International Space Station Program (ISSP) operations, and can also be used as a post -flight analysis tool. It is primarily used for situational awareness, and for contingency situations.

  15. Identification of clinical biomarkers for pre-analytical quality control of blood samples.

    Science.gov (United States)

    Kang, Hyun Ju; Jeon, Soon Young; Park, Jae-Sun; Yun, Ji Young; Kil, Han Na; Hong, Won Kyung; Lee, Mee-Hee; Kim, Jun-Woo; Jeon, Jae-Pil; Han, Bok Ghee

    2013-04-01

    Pre-analytical conditions are key factors in maintaining the high quality of biospecimens. They are necessary for accurate reproducibility of experiments in the field of biomarker discovery as well as achieving optimal specificity of laboratory tests for clinical diagnosis. In research at the National Biobank of Korea, we evaluated the impact of pre-analytical conditions on the stability of biobanked blood samples by measuring biochemical analytes commonly used in clinical laboratory tests. We measured 10 routine laboratory analytes in serum and plasma samples from healthy donors (n = 50) with a chemistry autoanalyzer (Hitachi 7600-110). The analyte measurements were made at different time courses based on delay of blood fractionation, freezing delay of fractionated serum and plasma samples, and at different cycles (0, 1, 3, 6, 9) of freeze-thawing. Statistically significant changes from the reference sample mean were determined using the repeated-measures ANOVA and the significant change limit (SCL). The serum levels of GGT and LDH were changed significantly depending on both the time interval between blood collection and fractionation and the time interval between fractionation and freezing of serum and plasma samples. The glucose level was most sensitive only to the elapsed time between blood collection and centrifugation for blood fractionation. Based on these findings, a simple formula (glucose decrease by 1.387 mg/dL per hour) was derived to estimate the length of time delay after blood collection. In addition, AST, BUN, GGT, and LDH showed sensitive responses to repeated freeze-thaw cycles of serum and plasma samples. These results suggest that GGT and LDH measurements can be used as quality control markers for certain pre-analytical conditions (eg, delayed processing or repeated freeze-thawing) of blood samples which are either directly used in the laboratory tests or stored for future research in the biobank.

  16. A content validated questionnaire for assessment of self reported venous blood sampling practices

    Directory of Open Access Journals (Sweden)

    Bölenius Karin

    2012-01-01

    Full Text Available Abstract Background Venous blood sampling is a common procedure in health care. It is strictly regulated by national and international guidelines. Deviations from guidelines due to human mistakes can cause patient harm. Validated questionnaires for health care personnel can be used to assess preventable "near misses"--i.e. potential errors and nonconformities during venous blood sampling practices that could transform into adverse events. However, no validated questionnaire that assesses nonconformities in venous blood sampling has previously been presented. The aim was to test a recently developed questionnaire in self reported venous blood sampling practices for validity and reliability. Findings We developed a questionnaire to assess deviations from best practices during venous blood sampling. The questionnaire contained questions about patient identification, test request management, test tube labeling, test tube handling, information search procedures and frequencies of error reporting. For content validity, the questionnaire was confirmed by experts on questionnaires and venous blood sampling. For reliability, test-retest statistics were used on the questionnaire answered twice. The final venous blood sampling questionnaire included 19 questions out of which 9 had in total 34 underlying items. It was found to have content validity. The test-retest analysis demonstrated that the items were generally stable. In total, 82% of the items fulfilled the reliability acceptance criteria. Conclusions The questionnaire could be used for assessment of "near miss" practices that could jeopardize patient safety and gives several benefits instead of assessing rare adverse events only. The higher frequencies of "near miss" practices allows for quantitative analysis of the effect of corrective interventions and to benchmark preanalytical quality not only at the laboratory/hospital level but also at the health care unit/hospital ward.

  17. A selective detection of lysophosphatidylcholine in dried blood spots for diagnosis of adrenoleukodystrophy by LC-MS/MS

    Directory of Open Access Journals (Sweden)

    Ryuichi Mashima

    2016-06-01

    Full Text Available X-linked adrenoleukodystrophy (X-ALD is a rare inherited metabolic disorder characterized by an impaired beta-oxidation of very long chain fatty acids in the peroxisomes. Recent studies have suggested that 1-hexacosanoyl-2-hydroxy-sn-glycero-3-phosphocholine (Lyso-PC 26:0 can be a sensitive biomarker for X-ALD. Although approximately 10-fold increase in the concentration of Lyso-PC 26:0 in DBSs from X-ALD-affected individuals were reported, whether the carriers might be distinguished from the healthy controls remained unclear. To address this question, we have validated previously developed LC-MS/MS-based analytical procedures using QC DBS. We found that the recovery of Lyso-PC 26:0 from the QC DBSs was 73.6 ± 0.3% when 2 μM of Lyso-PC 26:0 was spiked into the blood. Based on this result, the amounts of Lyso-PC 26:0 in the controls and ALD-affected individuals were 0.090 ± 0.004 (n = 11 and 1.078 ± 0.217 (n = 4 pmol/DBS, respectively. Interestingly, the concentration of Lyso-PC 26:0 in the carriers were 0.548 ± 0.095 pmol/DBS (n = 3, indicating that the carriers and the healthy controls can be distinguished. These results suggest that LC-MS/MS-based technique can be used for the detection of asymptomatic carriers and X-ALD-affected subjects in the newborn screening.

  18. Sampling methods to the statistical control of the production of blood components.

    Science.gov (United States)

    Pereira, Paulo; Seghatchian, Jerard; Caldeira, Beatriz; Santos, Paula; Castro, Rosa; Fernandes, Teresa; Xavier, Sandra; de Sousa, Gracinda; de Almeida E Sousa, João Paulo

    2017-12-01

    The control of blood components specifications is a requirement generalized in Europe by the European Commission Directives and in the US by the AABB standards. The use of a statistical process control methodology is recommended in the related literature, including the EDQM guideline. The control reliability is dependent of the sampling. However, a correct sampling methodology seems not to be systematically applied. Commonly, the sampling is intended to comply uniquely with the 1% specification to the produced blood components. Nevertheless, on a purely statistical viewpoint, this model could be argued not to be related to a consistent sampling technique. This could be a severe limitation to detect abnormal patterns and to assure that the production has a non-significant probability of producing nonconforming components. This article discusses what is happening in blood establishments. Three statistical methodologies are proposed: simple random sampling, sampling based on the proportion of a finite population, and sampling based on the inspection level. The empirical results demonstrate that these models are practicable in blood establishments contributing to the robustness of sampling and related statistical process control decisions for the purpose they are suggested for. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Analysis of multiple single nucleotide polymorphisms (SNP) on DNA traces from plasma and dried blood samples

    NARCIS (Netherlands)

    Catsburg, Arnold; van der Zwet, Wil C.; Morre, Servaas A.; Ouburg, Sander; Vandenbroucke-Grauls, Christina M. J. E.; Savelkoul, Paul H. M.

    2007-01-01

    Reliable analysis of single nucleotide polymorphisms (SNPs) in DNA derived from samples containing low numbers of cells or from suboptimal sources can be difficult. A new procedure to characterize multiple SNPs in traces of DNA from plasma and old dried blood samples was developed. Six SNPs in the

  20. Measurement of cerebral blood flow the blood sampling method using 99mTc-ECD. Simultaneous scintigram scanning of arterial blood samples and the brain with a gamma camera

    International Nuclear Information System (INIS)

    Hachiya, Takenori; Inugami, Atsushi; Iida, Hidehiro; Mizuta, Yoshihiko; Kawakami, Takeshi; Inoue, Minoru

    1999-01-01

    To measure regional cerebral blood flow (rCBF) by blood sampling using 99m Tc-ECD we devised a method of measuring the radioactive concentration in arterial blood sample with a gamma camera. In this method the head and a blood sample are placed within the same visual field to record the SPECT data of both specimens simultaneously. The results of an evaluation of the counting rate performance, applying the 30 hours decaying method using 99m Tc solution showed that this method is not comparable to the well-type scintillation counter and in clinical cases the active concentration in arterial blood sample remained well within the dynamic range. In addition, examination of the influence of scattered radiation from the brain by the dilution method showed that it was negligible at a distance of more than 7.5 cm between the brain and the arterial blood sample. In the present study we placed a head-shaped phantom next to the sample. The results of the examinations suggested that this method is suitable for clinical application, and because it does not require a well-type scintillation counter, it is expected to find wide application. (author)

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

    Science.gov (United States)

    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.

  2. Multispectral Imaging Analysis of Circulating Tumor Cells in Negatively Enriched Peripheral Blood Samples.

    Science.gov (United States)

    Miller, Brandon; Lustberg, Maryam; Summers, Thomas A; Chalmers, Jeffrey J

    2017-01-01

    A variety of biomarkers are present on cells in peripheral blood of patients with a variety of disorders, including solid tumor malignancies. While rare, characterization of these cells for specific protein levels with the advanced technology proposed, will lead to future validation studies of blood samples as "liquid biopsies" for the evaluation of disease status and therapeutic response. While circulating tumor cells (CTCs) have been isolated in the blood samples of patients with solid tumors, the exact role of CTCs as clinically useful predictive markers is still debated. Current commercial technology has significant bias in that a positive selection technology is used that preassumes specific cell surface markers (such as EpCAM) are present on CTCs. However, CTCs with low EpCAM expression have been experimentally demonstrated to be more likely to be missed by this method. In contrast, this application uses a previously developed, technology that performs a purely negative enrichment methodology on peripheral blood, yielding highly enriched blood samples that contain CTCs as well as other, undefined cell types. The focus of this contribution is the use of multispectral imaging of epifluorescent, microscopic images of these enriched cells in order to help develop clinically relevant liquid biopsies from peripheral blood samples.

  3. Temporal variability in urinary excretion of bisphenol A and seven other phenols in spot, morning, and 24-h urine samples

    DEFF Research Database (Denmark)

    Lassen, Tina Harmer; Frederiksen, Hanne; Jensen, Tina Kold

    2013-01-01

    (ICCs). More than 70% of the urine samples had detectable levels of BPA, triclosan (TCS), benzophenone-3 (BP-3) and sum of 2,4-dichlorophenol and 2,5-dichlorophenol (σDCP). We found low to moderate ICCs for BPA (0.10-0.42) and σDCP (0.39-0.72), whereas the ICCs for BP-3 (0.69-0.80) and TCS (0...

  4. Dealing with large sample sizes: comparison of a new one spot dot blot method to western blot.

    Science.gov (United States)

    Putra, Sulistyo Emantoko Dwi; Tsuprykov, Oleg; Von Websky, Karoline; Ritter, Teresa; Reichetzeder, Christoph; Hocher, Berthold

    2014-01-01

    Western blot is the gold standard method to determine individual protein expression levels. However, western blot is technically difficult to perform in large sample sizes because it is a time consuming and labor intensive process. Dot blot is often used instead when dealing with large sample sizes, but the main disadvantage of the existing dot blot techniques, is the absence of signal normalization to a housekeeping protein. In this study we established a one dot two development signals (ODTDS) dot blot method employing two different signal development systems. The first signal from the protein of interest was detected by horseradish peroxidase (HRP). The second signal, detecting the housekeeping protein, was obtained by using alkaline phosphatase (AP). Inter-assay results variations within ODTDS dot blot and western blot and intra-assay variations between both methods were low (1.04-5.71%) as assessed by coefficient of variation. ODTDS dot blot technique can be used instead of western blot when dealing with large sample sizes without a reduction in results accuracy.

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

    Directory of Open Access Journals (Sweden)

    Karen M. Tobias

    2016-11-01

    Full Text Available 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

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

    Science.gov (United States)

    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

  7. Stability and reliability of glycated haemoglobin measurements in blood samples stored at -20°C.

    Science.gov (United States)

    Venkataraman, Vijayachandrika; Anjana, Ranjit Mohan; Pradeepa, Rajendra; Deepa, Mohan; Jayashri, Ramamoorthy; Anbalagan, Viknesh Prabu; Akila, Bridgitte; Madhu, Sri Venkata; Lakshmy, Ramakrishnan; Mohan, Viswanathan

    2016-01-01

    To validate the stability of glycated haemoglobin (HbA1c) measurements in blood samples stored at -20°C for up to one month. The study group comprised 142 type 2 diabetic subjects visiting a tertiary centre for diabetes at Chennai city in south India. The HbA1c assay was done on a fasting blood sample using the Bio-Rad Variant machine on Day 0 (day of blood sample collection). Several aliquots were stored at -20°C and the assay was repeated on the 3rd, 7th, 15th, and 30th day after the sample collection. Bland-Altman plots were constructed and variation in the HbA1c levels on the different days was compared with the day 0 level. The median differences between HbA1c levels measured on Day 0 and the 3rd, 7th, 15th, and 30th day after blood collection were 0.0%, 0.2%, 0.3% and 0.5% respectively. Bland-Altman plot analysis showed that the differences between the day '0' and the different time points tend to get larger with time, but these were not clinically significant. HbA1c levels are relatively stable up to 2weeks, if blood samples are stored at -20°C. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. A simplified method for determination of radioactive iron in whole-blood samples

    DEFF Research Database (Denmark)

    Bukhave, Klaus; Sørensen, Anne Dorthe; Hansen, M.

    2001-01-01

    in humans. The overall recovery of radioiron from blood is more than 90%, and the coefficient of variation, as judged by the variation in the ratio Fe-55/Fe-59 is in the order of 4%. Combined with whole-body counting of 59Fe and direct gamma -counting of Fe-59 on blood samples, this method represents......For studies on iron absorption in man radioisotopes represent an easy and simple tool. However, measurement of the orbital electron emitting radioiron, Fe-55, in blood is difficult and insufficiently described in the literature. The present study describes a relatively simple method...... for simultaneous determination of Fe-55 and Fe-59 in blood, using a dry-ashing procedure and recrystallization of the remaining iron. The detection Limit of the method permits measurements of 0.1 Bq/ml blood thus allowing detection of Less than 1% absorption from a 40 kBq dose, which is ethically acceptable...

  9. Dark Spots

    Science.gov (United States)

    2006-01-01

    Dark spots (left) and 'fans' appear to scribble dusty hieroglyphics on top of the Martian south polar cap in two high-resolution Mars Global Surveyor, Mars Orbiter Camera images taken in southern spring. Each image is about 3-kilometers wide (2-miles).

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

    Science.gov (United States)

    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. © 2015 John Wiley & Sons Ltd.

  11. A comparison of various methods of blood sampling in mice and rats: Effects on animal welfare

    DEFF Research Database (Denmark)

    Harikrishnan, Vs; Hansen, Axel K; Abelson, Klas Sp

    2018-01-01

    -puncture activity and anxiety levels of rats and mice were measured using an elevated plus maze test and an open field test. Stress levels 24 h post-puncture were assessed by analysing faecal corticosteroid metabolites. Sucrose intake and faecal corticosteroid levels were not affected by the blood sampling...... procedures. Rats showed reduced activity in the open field test and an increased level of anxiety in the elevated plus maze test following retrobulbar plexus puncture and isoflurane anaesthesia. In mice, nest building activity was affected in all the groups compared with the control group, except for animals......This study was conducted to investigate the effects of blood sampling on animal welfare in a total of 60 NTac:SD rats and 72 C57BL/6NTac mice of both sexes. Blood was sampled either by sublingual vein puncture, tail vein puncture or by retrobulbar plexus/sinus puncture under light isoflurane...

  12. Radioimmunoassay screening and GC/MS confirmation of whole blood samples for drugs of abuse

    Energy Technology Data Exchange (ETDEWEB)

    Spiehler, V.R.; Sedgwick, P.

    From 1981 to 1984, an average of 300 radioimmunoassay screens on whole blood were performed each week in the authors laboratory. Most samples were screened for opiates phencyclidine and its analogs, barbiturates, and cocaine or its metabolite benzoylecgonine. A commercially available radioimmunoassay was used with modifications to facilitate screening of whole blood. Increasing sample size increased the sensitivity of the assay. Changing reagent concentration (1:1 dilution), incubation time, sample matrix (water, urine, or blood), or fraction counted (precipitate or supernatant) did not affect the utility of the standard curve or the sensitivity of the assay. All positive results for phencyclidine, opiates, cocaine, and related compounds were confirmed by GC/MA. Barbiturate positives were confirmed by UV spectrophotometry.

  13. EVALUATION OF ZEBU NELLORE CATTLE BLOOD SAMPLES USING THE CELL-DYN 3500 HEMATOLOGY ANALYZER

    Directory of Open Access Journals (Sweden)

    Alexandre Secorun Borges

    2014-12-01

    Full Text Available The Cell-dyn 3500 is a multiparameter flow cytometer, which may analyze samples from several species performing several simultaneous analyses. It is able to perform white blood cells, red blood cells and platelet counts, besides differential leukocyte counts, packed cell volume and hemoglobin determination. Cell-Dyn 3500 performs total leukocyte count both optically and by impedance. The equipment may choose one or other method, based on the reliability of the results. Erythrocyte and platelet counts are determined by impedance. Leukocyte differentiation is based on an optical principle, using separation in multiangular polarized light. The objective of this study was to compare the results of complete blood count of Zebu Nellore heifers from Celldyn 3500, with those obtained from a semi-automated cell counter (Celm CC 510 and the manual technique. Blood samples were collected from the jugular vein in 5 mL EDTA vacuum tubes from 58 Nellore heifers, at 24 months of age. Samples were processed in parallel in the three different techniques. Results were analyzed using paired t test, Pearson’s correlation and the Bland-Altmann method. There was a strong correlation for all parameters analyzed by Cell-Dyn 3500, manual method and semiautomated cell counter, except for basophils and monocytes counts. These results confirm that this analyzer is reliable for blood samples analysis of zebu cattle.

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

    Science.gov (United States)

    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.

  15. Rapid microbial sample preparation from blood using a novel concentration device.

    Directory of Open Access Journals (Sweden)

    Anna K Boardman

    Full Text Available 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 (MSSAto 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.

  16. Hybrid image and blood sampling input function for quantification of small animal dynamic PET data

    International Nuclear Information System (INIS)

    Shoghi, Kooresh I.; Welch, Michael J.

    2007-01-01

    We describe and validate a hybrid image and blood sampling (HIBS) method to derive the input function for quantification of microPET mice data. The HIBS algorithm derives the peak of the input function from the image, which is corrected for recovery, while the tail is derived from 5 to 6 optimally placed blood sampling points. A Bezier interpolation algorithm is used to link the rightmost image peak data point to the leftmost blood sampling point. To assess the performance of HIBS, 4 mice underwent 60-min microPET imaging sessions following a 0.40-0.50-mCi bolus administration of 18 FDG. In total, 21 blood samples (blood-sampled plasma time-activity curve, bsPTAC) were obtained throughout the imaging session to compare against the proposed HIBS method. MicroPET images were reconstructed using filtered back projection with a zoom of 2.75 on the heart. Volumetric regions of interest (ROIs) were composed by drawing circular ROIs 3 pixels in diameter on 3-4 transverse planes of the left ventricle. Performance was characterized by kinetic simulations in terms of bias in parameter estimates when bsPTAC and HIBS are used as input functions. The peak of the bsPTAC curve was distorted in comparison to the HIBS-derived curve due to temporal limitations and delay in blood sampling, which affected the rates of bidirectional exchange between plasma and tissue. The results highlight limitations in using bsPTAC. The HIBS method, however, yields consistent results, and thus, is a substitute for bsPTAC

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

    Science.gov (United States)

    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.

  18. Evaluation of preanalytical stability of the sample for complete blood count

    Directory of Open Access Journals (Sweden)

    Raghavendra Karanth P

    2011-09-01

    Full Text Available Objective: To evaluate the effect of time of incubation on complete blood count (CBC by using HmX analyzer. Methods: A cross sectional study was conducted at Manipal Acunova Limited, Bangalore. Ten blood samples which were stored at room temperature for CBC and differential count by using HmX analyzer were analyzed within one hour of draw and on 24, 48 and 72 hours of draw. Results: Differential count measured in an automated instrument changed over time. Conclusions: The finding of this study shows that some of CBC parameters can be changed with the incubation, therefore it is better to perform on a fresh sample.

  19. Application of gelatin zymography for evaluating low levels of contaminating neutrophils in red blood cell samples.

    Science.gov (United States)

    Achilli, Cesare; Ciana, Annarita; Balduini, Cesare; Risso, Angela; Minetti, Giampaolo

    2011-02-15

    Supposedly "homogeneous" red blood cell (RBC) samples are commonly obtained by "washing" whole blood free of plasma, platelets, and white cells with physiological solutions, a procedure that does not result, however, in sufficient removal of polymorphonuclear neutrophils (PMNs), leading to possible artifactual results. Pure RBC samples can be obtained only by leukodepletion procedures. Proposed here is a version of gelatin zymography adapted to detect matrix metalloproteinase 9 (MMP-9), selectively expressed by PMNs, in heterogeneous mixtures of RBCs and PMNs that can reveal contamination at levels as low as 1 PMN/10⁶ RBCs. Copyright © 2010 Elsevier Inc. All rights reserved.

  20. Eosinophilia in routine blood samples as a biomarker for solid tumor development

    DEFF Research Database (Denmark)

    Andersen, Christen Bertel L; Siersma, V.D.; Hasselbalch, H.C.

    2014-01-01

    eosinophilia in routine blood samples as a potential biomarker of solid tumor development in a prospective design. MATERIAL AND METHODS: From the Copenhagen Primary Care Differential Count (CopDiff) Database, we identified 356 196 individuals with at least one differential cell count (DIFF) encompassing...... was increased with mild eosinophilia [OR 1.93 (CI 1.29-2.89), p = 0.0013]. No associations with eosinophilia were observed for the remaining solid cancers. CONCLUSION: We demonstrate that eosinophilia in routine blood samples associates with an increased risk of bladder cancer. Our data emphasize...

  1. Blood venous sample collection: Recommendations overview and a checklist to improve quality.

    Science.gov (United States)

    Giavarina, Davide; Lippi, Giuseppe

    2017-07-01

    The extra-analytical phases of the total testing process have substantial impact on managed care, as well as an inherent high risk of vulnerability to errors which is often greater than that of the analytical phase. The collection of biological samples is a crucial preanalytical activity. Problems or errors occurring shortly before, or soon after, this preanalytical step may impair sample quality and characteristics, or else modify the final results of testing. The standardization of fasting requirements, rest, patient position and psychological state of the patient are therefore crucial for mitigating the impact of preanalytical variability. Moreover, the quality of materials used for collecting specimens, along with their compatibility, can guarantee sample quality and persistence of chemical and physical characteristics of the analytes over time, so safeguarding the reliability of testing. Appropriate techniques and sampling procedures are effective to prevent problems such as hemolysis, undue clotting in the blood tube, draw of insufficient sample volume and modification of analyte concentration. An accurate identification of both patient and blood samples is a key priority as for other healthcare activities. Good laboratory practice and appropriate training of operators, by specifically targeting collection of biological samples, blood in particular, may greatly improve this issue, thus lowering the risk of errors and their adverse clinical consequences. The implementation of a simple and rapid check-list, including verification of blood collection devices, patient preparation and sampling techniques, was found to be effective for enhancing sample quality and reducing some preanalytical errors associated with these procedures. The use of this tool, along with implementation of objective and standardized systems for detecting non-conformities related to unsuitable samples, can be helpful for standardizing preanalytical activities and improving the quality of

  2. Use of standard laboratory methods to obviate routine dithiothreitol treatment of blood samples with daratumumab interference.

    Science.gov (United States)

    Lintel, Nicholas J; Brown, Debra K; Schafer, Diane T; Tsimba-Chitsva, Farai M; Koepsell, Scott A; Shunkwiler, Sara M

    2017-01-01

    Daratumumab is an antibody currently used in the treatment of patients with refractory multiple myeloma. Blood samples from patients being treated with daratumumab may show panreactivity during pre-transfusion testing. To facilitate the provision of blood components for such patients, it is recommended that a baseline phenotype or genotype be established prior to starting treatment with daratumumab. If patient red blood cells (RBCs) require phenotyping after the start of daratumumab treatment, dithiothreitol (DTT) treatment of the patient's RBCs should be performed. The medical charts of four patients treated with daratumumab were reviewed. The individual number of doses ranged from 1 to 14; patient age ranged from 55 to 78 years; two men and two women were included in the review. Type and screen data were obtained from samples collected over 33 encounters with a range of 1 to 13 encounters per patient. All samples were tested initially by automated solid-phase testing. Any reactivity with solid phase led to tube testing with either low-ionic-strength saline, polyethylene glycol, or both. If incubation failed to eliminate the reactivity, the sample was sent to a reference laboratory for DTT treatment and phenotyping. Of the 33 samples tested, 23 (69.7%) samples had reactivity in solid-phase testing. In 8 of the 10 samples that did not react in solid-phase, testing was conducted more than four half-lives after the last dose of daratumumab. Of the 23 that had reactivity in solid-phase, 16 (69.6%) samples demonstrated loss of reactivity using common laboratory methods. For the seven patients whose sample reactivity was not initially eliminated, six were provided with phenotypically matched blood based on prior molecular testing. Only one sample was sent out for DTT treatment. These results suggest that daratumumab interference with pre-transfusion testing can be addressed using common laboratory methods. This finding could save time and money for laboratories that do

  3. Preanalytical aspects and sample quality assessment in metabolomics studies of human blood.

    Science.gov (United States)

    Yin, Peiyuan; Peter, Andreas; Franken, Holger; Zhao, Xinjie; Neukamm, Sabine S; Rosenbaum, Lars; Lucio, Marianna; Zell, Andreas; Häring, Hans-Ulrich; Xu, Guowang; Lehmann, Rainer

    2013-05-01

    Metabolomics is a powerful tool that is increasingly used in clinical research. Although excellent sample quality is essential, it can easily be compromised by undetected preanalytical errors. We set out to identify critical preanalytical steps and biomarkers that reflect preanalytical inaccuracies. We systematically investigated the effects of preanalytical variables (blood collection tubes, hemolysis, temperature and time before further processing, and number of freeze-thaw cycles) on metabolomics studies of clinical blood and plasma samples using a nontargeted LC-MS approach. Serum and heparinate blood collection tubes led to chemical noise in the mass spectra. Distinct, significant changes of 64 features in the EDTA-plasma metabolome were detected when blood was exposed to room temperature for 2, 4, 8, and 24 h. The resulting pattern was characterized by increases in hypoxanthine and sphingosine 1-phosphate (800% and 380%, respectively, at 2 h). In contrast, the plasma metabolome was stable for up to 4 h when EDTA blood samples were immediately placed in iced water. Hemolysis also caused numerous changes in the metabolic profile. Unexpectedly, up to 4 freeze-thaw cycles only slightly changed the EDTA-plasma metabolome, but increased the individual variability. Nontargeted metabolomics investigations led to the following recommendations for the preanalytical phase: test the blood collection tubes, avoid hemolysis, place whole blood immediately in ice water, use EDTA plasma, and preferably use nonrefrozen biobank samples. To exclude outliers due to preanalytical errors, inspect the biomarker signal intensities reflecting systematic as well as accidental and preanalytical inaccuracies before processing the bioinformatics data. © 2013 American Association for Clinical Chemistry.

  4. Comparative evaluation of blood and serum samples in rapid immunochromatographic tests for visceral leishmaniasis.

    Science.gov (United States)

    Kumar, Dinesh; Khanal, Basudha; Tiwary, Puja; Mudavath, Shyam Lal; Tiwary, Narendra K; Singh, Rupa; Koirala, Kanika; Boelaert, Marleen; Rijal, Suman; Sundar, Shyam

    2013-12-01

    Rapid diagnostic tests (RDTs) based on the detection of specific antibodies in serum are commonly used for the diagnosis of visceral leishmaniasis (VL). Several commercial kits are available, and some of them allow the use of whole-blood samples instead of serum. An RDT is much more user-friendly for blood samples than for serum samples. In this study, we examined the sensitivities and specificities of six different commercially available immunochromatographic tests for their accuracy in detecting Leishmania infection in whole blood and serum of parasitologically confirmed VL cases. This study was performed in areas of India and Nepal where VL is endemic. A total of 177 confirmed VL cases, 208 healthy controls from areas of endemicity (EHCs), 26 malaria patients (MP), and 37 tuberculosis (TB) patients were enrolled. The reproducibilities of the blood and serum results and between-reader and between-laboratory results were tested. In India, the sensitivities of all the RDTs ranged between 94.7 and 100.0%, with no significant differences between whole blood and serum. The specificities ranged between 92.4 and 100.0%, except for the specificity of the Onsite Leishmania Ab RevB kit, which was lower (33.6 to 42.0%). No differences in specificities were observed for blood and serum. In Nepal, the sensitivities of all the test kits, for whole-blood as well as serum samples, ranged between 96.3 and 100.0%, and the specificities ranged between 90.1 and 96.1%, again with the exception of that of the Onsite Leishmania Ab RevB test, which was markedly lower (48.7 to 49.3%). The diagnostic accuracies of all the tests, except for one brand, were excellent for the whole-blood and serum samples. We conclude that whole blood is an adequate alternative for serum in RDTs for VL, with sensitivities and specificities comparable to those obtained in serum samples, provided that the test kit is of overall good quality.

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

    Science.gov (United States)

    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

  6. 1-Hydroxypyrene Levels in Blood Samples of Rats After Exposure to Generator Fumes

    Science.gov (United States)

    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

  7. Detection of Merkel Cell Polyomavirus DNA in Serum Samples of Healthy Blood Donors

    Directory of Open Access Journals (Sweden)

    Elisa Mazzoni

    2017-11-01

    Full Text Available Merkel cell polyomavirus (MCPyV has been detected in 80% of Merkel cell carcinomas (MCC. In the host, the MCPyV reservoir remains elusive. MCPyV DNA sequences were revealed in blood donor buffy coats. In this study, MCPyV DNA sequences were investigated in the sera (n = 190 of healthy blood donors. Two MCPyV DNA sequences, coding for the viral oncoprotein large T antigen (LT, were investigated using polymerase chain reaction (PCR methods and DNA sequencing. Circulating MCPyV sequences were detected in sera with a prevalence of 2.6% (5/190, at low-DNA viral load, which is in the range of 1–4 and 1–5 copies/μl by real-time PCR and droplet digital PCR, respectively. DNA sequencing carried out in the five MCPyV-positive samples indicated that the two MCPyV LT sequences which were analyzed belong to the MKL-1 strain. Circulating MCPyV LT sequences are present in blood donor sera. MCPyV-positive samples from blood donors could represent a potential vehicle for MCPyV infection in receivers, whereas an increase in viral load may occur with multiple blood transfusions. In certain patient conditions, such as immune-depression/suppression, additional disease or old age, transfusion of MCPyV-positive samples could be an additional risk factor for MCC onset.

  8. Field evaluation of a broadly sensitive HIV-1 in-house genotyping assay for use with both plasma and dried blood spot specimens in a resource-limited country.

    Science.gov (United States)

    Inzaule, Seth; Yang, Chunfu; Kasembeli, Alex; Nafisa, Lillian; Okonji, Jully; Oyaro, Boaz; Lando, Richard; Mills, Lisa A; Laserson, Kayla; Thomas, Timothy; Nkengasong, John; Zeh, Clement

    2013-02-01

    HIV-1 drug resistance (HIVDR) assays are important tools in clinical management of HIV-infected patients on antiretroviral therapy (ART) and surveillance of drug-resistant variants at population levels. The high cost associated with commercial assays hinders their use in resource-limited settings. We adopted and validated a low-cost in-house assay using 68 matched plasma and dried blood spot (DBS) samples with a median viral load (VL) of 58,187 copies/ml, ranging from 253 to 3,264,850 against the commercial assay ViroSeq. Results indicated that the in-house assay not only had a higher plasma genotyping rate than did ViroSeq (94% versus 78%) but also was able to genotype 89.5% (51/57) of the matched DBS samples with VLs of ≥ 1,000 copies/ml. The sensitivity in detecting DR mutations by the in-house assay was 98.29% (95% confidence interval [CI], 97.86 to 98.72) on plasma and 96.54 (95% CI, 95.93 to 97.15) on DBS, and the specificity was 99.97% (95% CI, 99.91 to 100.00) for both sample types compared to ViroSeq. The minor DR mutation differences detected by the in-house assay against ViroSeq did not result in clinical significance. In addition, cost analysis showed that the in-house assay could reduce the genotyping cost by about 60% for both plasma and DBS compared to ViroSeq. This field condition evaluation highlights the potential utility of a cost-effective, subtype-independent, in-house genotyping assay using both plasma and DBS specimens for HIVDR clinical monitoring and population-based surveillance in resource-limited settings.

  9. Simultaneous determination of pentoxifylline, metabolites M1 (lisofylline), M4 and M5, and caffeine in plasma and dried blood spots for pharmacokinetic studies in preterm infants and neonates.

    Science.gov (United States)

    Page-Sharp, Madhu; Strunk, Tobias; Salman, Sam; Hibbert, Julie; Patole, Sanjay K; Manning, Laurens; Batty, Kevin T

    2017-11-30

    Advances in bioanalytical methods are facilitating micro-volume and dried blood spot (DBS) analysis of drugs in biological matrices for pharmacokinetic studies in children and neonates. We sought to develop a UPLC-MS/MS assay for simultaneous measurement of caffeine, pentoxifylline (PTX) and three metabolites of PTX in both plasma and DBS. Caffeine, PTX, the metabolites M1 (lisofylline), M4 and M5, and the internal standards (caffeine-d 9 and PTX-d 6 ) were separated using a Waters Aquity T3 UPLC C 18 column and gradient mobile phase (water-methanol-formic acid). Retention times for caffeine, M5, M4, PTX and M1 were 1.6, 1.7, 1.9, 2.0 and 2.1min, respectively, with a run time of 5min. The precision (≤10%) and accuracy (≤15%) across the concentration range 0.1-50mg/L for caffeine, PTX and the three metabolites in plasma and DBS were within accepted limits, as were the limits of quantification (100μg/L for caffeine and 10μg/L for PTX, M1, M4 and M5). Caffeine, PTX and the metabolites were stable in DBS for >34days at room and refrigerated temperatures. Plasma and DBS samples were obtained from 24 preterm infants recruited into a clinical pharmacokinetic study of PTX. Paired analysis indicated that DBS concentrations were 9% lower than concurrent plasma concentrations for caffeine, 7% lower for PTX (consistent with the blood:plasma ratio) and 13% lower for M1 (lisofylline). The validated UPLC-MS/MS method is suitable for micro-volume plasma and DBS analysis of caffeine, PTX and its metabolites for pharmacokinetic studies in paediatric patients. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. The effect of introducing point-of-care or dried blood spot analysis on the uptake of hepatitis C virus testing in high-risk populations: A systematic review of the literature.

    Science.gov (United States)

    Coats, Josh T; Dillon, John F

    2015-11-01

    Testing for hepatitis C virus (HCV) infection typically relies upon blood samples taken by traditional phlebotomy for laboratory processing. Novel testing methods, including using dried blood spots (DBS) and point-of-care (PoC) testing enable easier access to high risk populations who have less frequent contact with healthcare professionals. Many of these individuals have been exposed to HCV but have not previously been tested. We aimed to establish whether the availability of these novel testing methods increased either uptake of testing or the number of new diagnoses of HCV. The PubMed, Cochrane and SCOPUS databases were searched for terms relating to the study. References and associated bibliographies were also examined for further relevant articles. Studies were included if they contained quantitative data on frequency of testing and/or new diagnoses following the introduction of PoC and/or DBS testing of high-risk populations. Studies were then examined for findings and limitations and graded upon the quality of evidence provided. No studies were found which introduced PoC testing and determined its effect on frequency of testing or new diagnoses. Six studies were identified in which DBS testing was introduced and its effect evaluated. Two of the studies were randomised controlled trials, two were prospective cohort studies, one was an ecological study and one was a clinical audit. Populations studied included those attending substance misuse clinics, prisons and needle exchanges. Injection drug use was the commonest risk factor for HCV. Five of the six studies provided evidence that the introduction of DBS testing increased the number of tests, new diagnoses or both. Current evidence indicates that DBS testing availability may increase the uptake of testing for HCV in high-risk populations. There is currently no evidence regarding the efficacy of PoC testing in these populations. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. A pilot study using residual newborn dried blood spots to assess the potential role of cytomegalovirus and Toxoplasma gondii in the etiology of congenital hydrocephalus.

    Science.gov (United States)

    Simeone, Regina M; Rasmussen, Sonja A; Mei, Joanne V; Dollard, Sheila C; Frias, Jaime L; Shaw, Gary M; Canfield, Mark A; Meyer, Robert E; Jones, Jeffrey L; Lorey, Fred; Honein, Margaret A

    2013-07-01

    Congenital hydrocephalus is a condition characterized by accumulation of cerebrospinal fluid in the ventricles of the brain. Prenatal infections are risk factors for some birth defects. This pilot study investigated whether residual dried blood spots (DBS) could be used to assess infections as risk factors for birth defects by examining the associations between prenatal infection with Toxoplasma gondii (T. gondii) or cytomegalovirus (CMV) with congenital hydrocephalus. Case-infants with hydrocephalus (N=410) were identified among live-born infants using birth defects surveillance systems in California, North Carolina, and Texas. Control-infants without birth defects were randomly selected from the same geographic areas and time periods as case-infants (N=448). We tested residual DBS from case- and control-infants for T. gondii immunoglobulin M and CMV DNA. When possible, we calculated crude odds ratios (cORs) and confidence intervals (CIs). Evidence for prenatal T. gondii infection was more common among case-infants (1.2%) than control-infants (0%; p=0.11), and evidence for prenatal CMV infection was higher among case-infants (1.5%) than control-infants (0.7%; cOR: 2.3; 95% CI: 0.48, 13.99). Prenatal infections with T. gondii and CMV occurred more often among infants with congenital hydrocephalus than control-infants, although differences were not statistically significant. This pilot study highlighted some challenges in using DBS to examine associations between certain infections and birth defects, particularly related to reduced sensitivity and specimen storage conditions. Further study with increased numbers of specimens and higher quality specimens should be considered to understand better the contribution of these infections to the occurrence of congenital hydrocephalus. Copyright © 2013 Wiley Periodicals, Inc.

  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.

    Science.gov (United States)

    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 (blood, prolonged direct contact between infected and healthy cattle may be considered as a risk factor for BLV transmission. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Large-scale prospective T cell function assays in shipped, unfrozen blood samples

    DEFF Research Database (Denmark)

    Hadley, David; Cheung, Roy K; Becker, Dorothy J

    2014-01-01

    , for measuring core T cell functions. The Trial to Reduce Insulin-dependent diabetes mellitus in the Genetically at Risk (TRIGR) type 1 diabetes prevention trial used consecutive measurements of T cell proliferative responses in prospectively collected fresh heparinized blood samples shipped by courier within...... cell immunocompetence. We have found that the vast majority of the samples were viable up to 3 days from the blood draw, yet meaningful responses were found in a proportion of those with longer travel times. Furthermore, the shipping time of uncooled samples significantly decreased both the viabilities...... North America. In this article, we report on the quality control implications of this simple and pragmatic shipping practice and the interpretation of positive- and negative-control analytes in our assay. We used polyclonal and postvaccination responses in 4,919 samples to analyze the development of T...

  14. Possibilities of radioisotope renal clearances without blood sampling in patients with chronic glomerulonephritis

    International Nuclear Information System (INIS)

    Shejretova, E.; Velikov, P.

    1987-01-01

    Two clearance methods are proposed: 1) Effective renal plasma flow determination with 131 J-hippuran, and 2) glomeruli filtration determination with 169 Yb-EDTA. Thirty one adult patients with chronic glomerulonephritis and 59 without clinical and laboratory evidence of urinary tract disease comprised the study group. Statistically significant decrease in the effective renal plasma flow (p<0,001) and of glomerular filtration (p<0,01) was recorded in comparison to the control group, which is an evidence of the good diagnostic possibilities of the methods. The significant difference between the group of patients with elevated blood urea levels or pathologic Zimnincki's test and reduced glomerular filtration, compared to those with normal glomerular filtration, is an indication for the high diagnostic value of the glomerular filtration method. The clearance methods without blood sampling are particularly useful in the diagnosis and follow-up of patients with chronic glomerulonephritis, because of the burdenless technique, lack of blood sampling, and sufficient accuracy

  15. Development and validation of an HPLC-MS/MS method for the analysis of dexamethasone from pig synovial fluid using dried matrix spotting.

    Science.gov (United States)

    Christianson, Chad D; Laine, Derek F; Zimmer, Jennifer Sd; Johnson, Casey Jl; Sheaff, Chrystal N; Carpenter, Anna; Needham, Shane R

    2010-11-01

    Dried matrix spot techniques were employed to validate an HPLC-MS/MS assay for the determination of dexamethasone in clear Yorkshire pig synovial fluid using 15 µl of sample. We have adopted the term dried matrix spot to indicate that the techniques used for dried blood spots can be applied to nonblood matrices. The dried matrix spot method employs a color-indicating process developed at Alturas Analytics that enhances the ability to analyze transparent fluids spotted onto collection paper by allowing the analyst to visually verify the location of the dried sample spot. The method was shown to be accurate (±4.3%) and precise (14.2% at the LLOQ and ≤10.0% at all other concentrations) across the dynamic range of the assay. The method shows the potential application of dried matrix spot techniques for the analysis of transparent biological fluids.

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

    Science.gov (United States)

    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. Dried Blood Spots, an Affordable Tool to Collect, Ship, and Sequence gDNA from Patients with an X-Linked Agammaglobulinemia Phenotype Residing in a Developing Country

    Directory of Open Access Journals (Sweden)

    Gesmar R. S. Segundo

    2018-02-01

    Full Text Available BackgroundNew sequencing techniques have revolutionized the identification of the molecular basis of primary immunodeficiency disorders (PID not only by establishing a gene-based diagnosis but also by facilitating defect-specific treatment strategies, improving quality of life and survival, and allowing factual genetic counseling. Because these techniques are generally not available for physicians and their patients residing in developing countries, collaboration with overseas laboratories has been explored as a possible, albeit cumbersome, strategy. To reduce the cost of time and temperature-sensitive shipping, we selected Guthrie cards, developed for newborn screening, to collect dried blood spots (DBS, as a source of DNA that can be shipped by regular mail at minimal cost.MethodBlood was collected and blotted onto the filter paper of Guthrie cards by completely filling three circles. We enrolled 20 male patients with presumptive X-linked agammaglobulinemia (XLA cared for at the Vietnam National Children’s Hospital, their mothers, and several sisters for carrier analysis. DBS were stored at room temperature until ready to be shipped together, using an appropriately sized envelope, to a CLIA-certified laboratory in the US for sequencing. The protocol for Sanger sequencing was modified to account for the reduced quantity of gDNA extracted from DBS.ResultHigh-quality gDNA could be extracted from every specimen. Bruton tyrosine kinase (BTK mutations were identified in 17 of 20 patients studied, confirming the diagnosis of XLA in 85% of the study cohort. Type and location of the mutations were similar to those reported in previous reviews. The mean age when XLA was suspected clinically was 4.6 years, similar to that reported by Western countries. Two of 15 mothers, each with an affected boy, had a normal BTK sequence, suggesting gonadal mosaicism.ConclusionDBS collected on Guthrie cards can be shipped inexpensively by airmail across continents

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

    Science.gov (United States)

    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

    2017-12-01

    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.

  19. Blood Sampling Seasonality as an Important Preanalytical Factor for Assessment of Vitamin D Status

    Directory of Open Access Journals (Sweden)

    Bonelli Patrizia

    2016-04-01

    Full Text Available 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.

  20. [Automated serial diagnosis of donor blood samples. Ergonomic and economic organization structure].

    Science.gov (United States)

    Stoll, T; Fischer-Fröhlich, C L; Mayer, G; Hanfland, P

    1990-01-01

    A comprehensive computer-aided administration-system for blood-donors is presented. Ciphered informations of barcode-labels allow the automatic and nevertheless selective pipetting of samples by pipetting-robots. Self-acting analysis-results are transferred to a host-computer in order to actualize a donor data-base.

  1. Uranium concentration in blood samples of Southern Iraqi leukemia patients using CR-39 track detector

    International Nuclear Information System (INIS)

    Al-Hamzawi, A.A.; Al-Qadisiyah University, Qadisiyah; Jaafar, M.S.; Tawfiq, N.F.

    2014-01-01

    The simple and effective technique of fission track etch has been applied to determine trace concentration of uranium in human blood samples taken from two groups of male and female participants: leukemia patients and healthy subjects group. The blood samples of leukemia patients and healthy subjects were collected from three key southern governorates namely, Basrah, Muthanna and Dhi-Qar. These governorates were the centers of intensive military activities during the 1991 and 2003 Gulf wars, and the discarded weapons are still lying around in these regions. CR-39 track detector was used for registration of induced fission tracks. The results show that the highest recorded uranium concentration in the blood samples of leukemia patients was 4.71 ppb (female, 45 years old, from Basrah) and the minimum concentration was 1.91 ppb (male, 3 years old, from Muthanna). For healthy group, the maximum uranium concentration was 2.15 ppb (female, 55 years old, from Basrah) and the minimum concentration was 0.86 ppb (male, 5 years old, from Dhi-Qar). It has been found that the uranium concentrations in human blood samples of leukemia patients are higher than those of the healthy group. These uranium concentrations in the leukemia patients group were significantly different (P < 0.001) from those in the healthy group. (author)

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

    CSIR Research Space (South Africa)

    Smith, S

    2015-02-01

    Full Text Available 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...

  3. Workup of Human Blood Samples for Deep Sequencing of HIV-1 Genomes

    NARCIS (Netherlands)

    Cornelissen, Marion; Gall, Astrid; van der Kuyl, Antoinette; Wymant, Chris; Blanquart, François; Fraser, Christophe; Berkhout, Ben

    2018-01-01

    We describe a detailed protocol for the manual workup of blood (plasma/serum) samples from individuals infected with the human immunodeficiency virus type 1 (HIV-1) for deep sequence analysis of the viral genome. The study optimizing the assay was performed in the context of the BEEHIVE (Bridging

  4. Standardised Resting Time Prior to Blood Sampling and Diurnal Variation Associated with Risk of Patient Misclassification

    DEFF Research Database (Denmark)

    Bøgh Andersen, Ida; Brasen, Claus L.; Christensen, Henry

    2015-01-01

    BACKGROUND: According to current recommendations, blood samples should be taken in the morning after 15 minutes' resting time. Some components exhibit diurnal variation and in response to pressures to expand opening hours and reduce waiting time, the aims of this study were to investigate the imp...

  5. Development of a One-Step Probe Based Molecular Assay for Rapid Immunodiagnosis of Infection with M. tuberculosis Using Dried Blood Spots

    DEFF Research Database (Denmark)

    Blauenfeldt, Thomas; Heyckendorf, Jan; Graff Jensen, Sidse

    2014-01-01

    detection. AIM: To develop a robust IP-10 based molecular assay for the diagnosis of infection with M. tubercuolsis from whole blood and DBS. METHOD: We developed a one-step probe based multiplex RT-qPCR assay for detecting IP-10 and IFN-γ mRNA expression from whole blood and DBS samples. The assay.......2, IQR 10.7-67.0) and persons with latent tuberculosis infection (LTBI) (41.2, IQR 9.8-64.9) compared to healthy controls (1.6, IQR 1.1-2.4; pdiagnostic accuracy to QFT-TB, sensitivity (85% and 88% vs 85%) and specificity (96% and 96% vs 97...

  6. A blood sampling microsystem for pharmacokinetic applications: design, fabrication, and initial results.

    Science.gov (United States)

    Li, Tao; Barnett, Adam; Rogers, Karen L; Gianchandani, Yogesh B

    2009-12-21

    This paper describes a microsystem for automated blood sampling from laboratory mice used in pharmacokinetic studies. Intended to be mounted as a "backpack" on a mouse, it uses a microneedle, reservoir, and an actuator to instantaneously prick the animal for a time-point sample, eliminating the need for a tethered catheter with large dead volume. The blood is collected by capillary effect through a 31-33 gauge microneedle (250-210 microm OD) into a approximately 1 microL micromachined steel reservoir. The voice coil actuator provides a peak force of approximately 300 mN, which amply exceeds the measured piercing force of mouse skin (i.e., 60-85 mN for a 31-gauge needle with 12 degrees bevel). The sampling system was tested in vitro using a mock vessel with adjustable pressure; the reservoir was filled in pressure. The system may also be used to sample interstitial fluid, but the absence of blood pressure makes it necessary to enhance the capillary effect of the needle. This is accomplished by either electropolishing the inner surface to make it more hydrophilic or using a polymer wire insert to increase the surface area. The steel surface of the reservoir is also coated with silicon oxynitride by plasma-enhanced chemical vapor deposition to improve its hydrophilicity. Blood from fresh bovine tissue was collected into the reservoir to simulate interstitial fluid sampling. In vivo tests on live, anesthetized mice resulted in successful collection of blood into the reservoir. The possible integration of the device in microanalytical systems and the device scalability for multisampling are discussed.

  7. Comparative Analysis of Clinical Samples Showing Weak Serum Reaction on AutoVue System Causing ABO Blood Typing Discrepancies

    OpenAIRE

    Jo, Su Yeon; Lee, Ju Mi; Kim, Hye Lim; Sin, Kyeong Hwa; Lee, Hyeon Ji; Chang, Chulhun Ludgerus; Kim, Hyung-Hoi

    2016-01-01

    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...

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    Williams Adam R

    2009-12-01

    Full Text Available Abstract 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

  10. A simple method for regional cerebral blood flow measurement by one-point arterial blood sampling and 123I-IMP microsphere model (part 2). A study of time correction of one-point blood sample count

    International Nuclear Information System (INIS)

    Masuda, Yasuhiko; Makino, Kenichi; Gotoh, Satoshi

    1999-01-01

    In our previous paper regarding determination of the regional cerebral blood flow (rCBF) using the 123 I-IMP microsphere model, we reported that the accuracy of determination of the integrated value of the input function from one-point arterial blood sampling can be increased by performing correction using the 5 min: 29 min ratio for the whole-brain count. However, failure to carry out the arterial blood collection at exactly 5 minutes after 123 I-IMP injection causes errors with this method, and there is thus a time limitation. We have now revised out method so that the one-point arterial blood sampling can be performed at any time during the interval between 5 minutes and 20 minutes after 123 I-IMP injection, with addition of a correction step for the sampling time. This revised method permits more accurate estimation of the integral of the input functions. This method was then applied to 174 experimental subjects: one-point blood samples collected at random times between 5 and 20 minutes, and the estimated values for the continuous arterial octanol extraction count (COC) were determined. The mean error rate between the COC and the actual measured continuous arterial octanol extraction count (OC) was 3.6%, and the standard deviation was 12.7%. Accordingly, in 70% of the cases, the rCBF was able to be estimated within an error rate of 13%, while estimation was possible in 95% of the cases within an error rate of 25%. This improved method is a simple technique for determination of the rCBF by 123 I-IMP microsphere model and one-point arterial blood sampling which no longer shows a time limitation and does not require any octanol extraction step. (author)

  11. Midazolam sedates Passeriformes for field sampling but affects multiple venous blood analytes

    Directory of Open Access Journals (Sweden)

    Heatley JJ

    2015-01-01

    Full Text Available J Jill Heatley,1 Jennifer Cary,2,3 Lyndsey Kingsley,1 Hughes Beaufrere,4 Karen E Russell,5 Gary Voelker2,3 1Department of Small Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, 2Department of Wildlife and Fisheries Sciences, 3Texas A&M Biodiversity Research and Teaching Collections, Texas A&M University, College Station, TX, USA; 4Health Sciences Centre, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada; 5Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, College Station, TX, USA Abstract: Feasibility and effect of midazolam administration on blood analytes and for sedation of Passeriformes being collected in a larger study of genetic biodiversity was assessed. Midazolam (5.6±2.7 mg/kg was administered intranasally prior to sampling, euthanasia, and specimen preparation of 104 passerine birds. Each bird was assessed for sedation score and then multiple analytes were determined from jugular blood samples using the i-STAT® point of care analyzer at “bird side”. Most birds were acceptably sedated, sedation became more pronounced as midazolam dose increased, and only a single bird died. Electrolyte concentrations and venous blood gas analytes were affected by midazolam administration while blood pH, packed cell volume, hemoglobin, and calculated hematocrit were not. Intranasal midazolam gives adequate sedation and is safe for short-term use in free-living Passeriformes. Based on venous blood analyte data, sedation of Passeriformes prior to handling appears to reduce stress but also produces venous blood gas differences consistent with hypoventilation relative to birds which were not given midazolam. Further study is recommended to investigate midazolam's continued use in free-living avian species. Studies should include safety, reversal and recovery, effect upon additional endogenous analytes, and compatibility with studies of ecology and toxicology

  12. Fabrication and Characterization of a Microfluidic Device to Ultrapurify Blood Samples

    KAUST Repository

    Tallerico, Marco

    2015-05-04

    The improvement of blood cell sorting techniques in recent years have attracted the attention of many researchers due to the possible benefits that these methods can lead in biology, regenerative medicine, materials science and therapeutic area. In this work a cell sorting technique based on filtration is described. The separation occurs by means of a microfluidic device, suitably designed, manufactured and tested, that is connected to an external experimental set-up. The fabrication process can be divided in two parts: at first it is described the manufacturing process of a filtering membrane, with holes of specific size that allow the passage of only certain cell types. Following the microfluidic device is fabricated through the mechanical micromilling. The membrane and the microdevice are suitably bonded and tested by means of an external connection with syringe pumps that inject blood samples at specific flow rates. The device is designed to separate blood cells and tumor cells only by using differences in size and shape. In particular during the first experiments red blood cells and platelets are sorted from white blood cells; in the other experiments red blood cells and platelets are separated from white blood cells and tumor cells. The microdevice has proven to be very efficient, in fact a capture efficiency of 99% is achieved. For this reason it could be used in identification and isolation of circulating tumor cells, a very rare cancer cell type whose presence in the bloodstream could be symptom of future solid tumor formation. The various experiments have also demonstrated that tumor cells survive even after the separation treatment, and then the suffered stress during the sorting process does not harm the biological sample.

  13. Using improved serial blood sampling method of mice to study pharmacokinetics and drug-drug interaction.

    Science.gov (United States)

    Watanabe, Ayahisa; Watari, Ryosuke; Ogawa, Keiko; Shimizu, Ryosuke; Tanaka, Yukari; Takai, Nozomi; Nezasa, Ken-ichi; Yamaguchi, Yoshitaka

    2015-03-01

    In pharmacokinetic evaluation of mice, using serial sampling methods rather than a terminal blood sampling method could reduce the number of animals needed and lead to more reliable data by excluding individual differences. In addition, using serial sampling methods can be valuable for evaluation of the drug-drug interaction (DDI) potential of drug candidates. In this study, we established an improved method for serially sampling the blood from one mouse by only one incision of the lateral tail vein, and investigated whether our method could be adapted to pharmacokinetic and DDI studies. After intravenous and oral administration of ibuprofen and fexofenadine (BCS class II and III), the plasma concentration and pharmacokinetic parameters were evaluated by our method and a terminal blood sampling method, with the result that both methods gave comparable results (ibuprofen: 63.8 ± 4.0% and 64.4%, fexofenadine: 6.5 ± 0.7% and 7.9%, respectively, in bioavailability). In addition, our method could be adapted to DDI study for cytochrome P450 and organic anion transporting polypeptide inhibition. These results demonstrate that our method can be useful for pharmacokinetic evaluation from the perspective of reliable data acquisition as well as easy handling and low stress to mice and improve the quality of pharmacokinetic and DDI studies. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  14. Alternative sampling site for blood glucose testing in cats: giving the ears a rest.

    Science.gov (United States)

    Zeugswetter, Florian K; Rebuzzi, Laura; Karlovits, Sonja

    2010-09-01

    BACKGROUND AND STUDY RATIONALE: Home monitoring is an important part of the long-term management of diabetic cats. Despite the extensive use of glucometers in this species, up until now only the pinna of the ear has been validated as a testing site. This cross-sectional study investigated the feasibility and validity of sampling from the metacarpal/metatarsal pads in hospitalised cats with various diseases. The large pads were compared with the ear as a sampling site in 75 cats. Lancing the pads was tolerated very well. If the initial drop of blood was too small, an adequate volume of blood was almost always achieved by squeezing the pads. No significant differences were observed in first-attempt success rate or glucose values between the two sites. Due to the inability to obtain an adequate volume of blood or struggling, no measurement was possible in four cats. While further work is necessary to assess the utility of this technique, especially in the home environment, the results indicate that the metacarpal pads, in particular, may offer a viable alternative testing site for the measurement of blood glucose concentrations, especially if ear sampling fails. Copyright 2010 ISFM and AAFP. Published by Elsevier Ltd. All rights reserved.

  15. Determination of Se and Zn elements in blood serum samples by neutron activation analysis

    International Nuclear Information System (INIS)

    Indah Kusmartini; Rukruk Rukayah; Woro Yatu NS; Muhayatun; Rochestry Sofyan

    2010-01-01

    Se and Zn are essential elements being required for activity several enzyme systems in human metabolism. The elements in blood serum as well as to obtain information regarding the health status of individuals, so that important to investigated. Commonly Se and Zn elements in blood serum are low in concentration with limited samples weight, therefore high sensitive and accurate analysis method like NAA are needed. This study aims to determine the content of the elements Se and Zn in blood serum of employee using NAA technique. The samples were freeze dried then irradiated at rabbit system facility of G.A. Siwabessy Serpong reactor with neutron flux ~10 13 n.cm -2 .s -1 for 2 hours. Then samples were counted for 50000 s by HPGe spectrometer gamma and analyzed by software GENIE 2000. Method validation was also applied by analyzing the Reference Material IAEA Animal Blood RM A-13. The range concentration of elements Se and Zn were 0.06-0.17 μg/mL and 0.56-1.37 μg/mL with overall mean 0.10 ± 0.01 μg/mL and 0.97 ± 0.07 μg/mL. The value showed that appropriate with last researcher and another country. (author)

  16. Interlaboratory comparison of PCR-based identification of Candida and Aspergillus DNA in spiked blood samples.

    Science.gov (United States)

    Reichard, Utz; Buchheidt, Dieter; Lass-Flörl, Cornelia; Loeffler, Juergen; Lugert, Raimond; Ruhnke, Markus; Tintelnot, Kathrin; Weig, Michael; Groß, Uwe

    2012-09-01

    Despite PCR per se being a powerful and sensitive technique, regarding the detection of fungi in patients' blood, no consensus for a standardised PCR protocol yet exists. To complement other ongoing or accomplished studies which tackle this problem, the German Reference Center for Systemic Mycoses conducted an interlaboratory comparison starting with blood samples spiked with fungal cell elements. Altogether, six laboratories using in-house PCR-protocols from Germany and Austria participated in the trial. Blood samples were spiked with vital cells of Candida albicans or Aspergillus fumigatus. Candida was used in the yeast form, whereas Aspergillus cells were either spiked as conidia or as very young germlings, also known as smoo cells. Spiked blood samples contained between 10 and 10 000 cells ml(-1). Depending on the techniques used for fungal cell disruption and DNA-amplification, detection quality was variable between laboratories, but also differed within single laboratories in different trials particularly for samples spiked with less than 100 cells ml(-1). Altogether, at least regarding the detection of A. fumigatus, two of six laboratories showed constant reliable test results also with low fungal cell number spiked samples. Protocols used by these labs do not differ substantially from others. However, as particularities, one protocol included a conventional phenol chloroform extraction during the DNA preparation process and the other included a real time PCR-protocol based on FRET probes. Other laboratory comparisons on the basis of clinical samples should follow to further evaluate the procedures. The difficulties and problems of such trials in general are discussed. © 2012 Blackwell Verlag GmbH.

  17. Suitability of small diagnostic peripheral-blood samples for cell-therapy studies.

    Science.gov (United States)

    Stephanou, Coralea; Papasavva, Panayiota; Zachariou, Myria; Patsali, Petros; Epitropou, Marilena; Ladas, Petros; Al-Abdulla, Ruba; Christou, Soteroulla; Antoniou, Michael N; Lederer, Carsten W; Kleanthous, Marina

    2017-02-01

    Primary hematopoietic stem and progenitor cells (HSPCs) are key components of cell-based therapies for blood disorders and are thus the authentic substrate for related research. We propose that ubiquitous small-volume diagnostic samples represent a readily available and as yet untapped resource of primary patient-derived cells for cell- and gene-therapy studies. In the present study we compare isolation and storage methods for HSPCs from normal and thalassemic small-volume blood samples, considering genotype, density-gradient versus lysis-based cell isolation and cryostorage media with different serum contents. Downstream analyses include viability, recovery, differentiation in semi-solid media and performance in liquid cultures and viral transductions. We demonstrate that HSPCs isolated either by ammonium-chloride potassium (ACK)-based lysis or by gradient isolation are suitable for functional analyses in clonogenic assays, high-level HSPC expansion and efficient lentiviral transduction. For cryostorage of cells, gradient isolation is superior to ACK lysis, and cryostorage in freezing media containing 50% fetal bovine serum demonstrated good results across all tested criteria. For assays on freshly isolated cells, ACK lysis performed similar to, and for thalassemic samples better than, gradient isolation, at a fraction of the cost and hands-on time. All isolation and storage methods show considerable variation within sample groups, but this is particularly acute for density gradient isolation of thalassemic samples. This study demonstrates the suitability of small-volume blood samples for storage and preclinical studies, opening up the research field of HSPC and gene therapy to any blood diagnostic laboratory with corresponding bioethics approval for experimental use of surplus material. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    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

  19. Mongolian spots

    Directory of Open Access Journals (Sweden)

    Divya Gupta

    2013-01-01

    Full Text Available Mongolian spots (MS are birthmarks that are present at birth and their most common location is sacrococcygeal or lumbar area. Lesions may be single or multiple and usually involve < 5% total body surface area. They are macular and round, oval or irregular in shape. The color varies from blue to greenish, gray, black or a combination of any of the above. The size varies from few to more than 20 centimetres. Pigmentation is most intense at the age of one year and gradually fades thereafter. It is rarely seen after the age of 6 years. Aberrant MS over occiput, temple, mandibular area, shoulders and limbs may be confused with other dermal melanocytoses and bruises secondary to child abuse, thus necessitating documentation at birth. Although regarded as benign, recent data suggest that MS may be associated with inborn errors of metabolism and neurocristopathies. Mongolian spots usually resolve by early childhood and hence no treatment is generally needed if they are located in the sacral area. However, sometimes it may be required for extrasacral lesions for cosmesis.

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

    Science.gov (United States)

    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

  1. Gene methylation parallelisms between peripheral blood cells and oral mucosa samples in relation to overweight.

    Science.gov (United States)

    San-Cristobal, Rodrigo; Navas-Carretero, Santiago; Milagro, Fermín I; Riezu-Boj, J Ignacio; Guruceaga, Elizabeth; Celis-Morales, Carlos; Livingstone, Katherine M; Brennan, Lorraine; Lovegrove, Julie A; Daniel, Hannelore; Saris, Wim H; Traczyk, Iwonna; Manios, Yannis; Gibney, Eileen R; Gibney, Michael J; Mathers, John C; Martinez, J Alfredo

    2016-08-01

    Epigenetics has an important role in the regulation of metabolic adaptation to environmental modifications. In this sense, the determination of epigenetic changes in non-invasive samples during the development of metabolic diseases could play an important role in the procedures in primary healthcare practice. To help translate the knowledge of epigenetics to public health practice, the present study aims to explore the parallelism of methylation levels between white blood cells and buccal samples in relation to obesity and associated disorders. Blood and buccal swap samples were collected from a subsample of the Spanish cohort of the Food4Me study. Infinium HumanMethylation450 DNA Analysis was carried out for the determination of methylation levels. Standard deviation for β values method and concordance correlation analysis were used to select those CpG which showed best parallelism between samples. A total of 277 CpGs met the criteria and were selected for an enrichment analysis and a correlation analysis with anthropometrical and clinical parameters. From those selected CpGs, four presented high associations with BMI (cg01055691 in GAP43; r = -0.92 and rho = -0.84 for blood; r = -0.89 and rho = -0.83 for buccal sample), HOMA-IR (cg00095677 in ATP2A3; r = 0.82 and rho = -0.84 for blood; r = -0.8 and rho = -0.83 for buccal sample) and leptin (cg14464133 in ADARB2; r = -0.9182 and rho = -0.94 for blood; r = -0.893 and rho = -0.79 for buccal sample). These findings demonstrate the potential application of non-invasive buccal samples in the identification of surrogate epigenetic biomarkers and identify methylation sites in GAP43, ATP2A3 and ADARB2 genes as potential targets in relation to overweight management and insulin sensibility.

  2. Highly Effective DNA Extraction Method from Fresh, Frozen, Dried and Clotted Blood Samples

    Directory of Open Access Journals (Sweden)

    Jaleh Barar

    2011-09-01

    Full Text Available Introduction: Today, with the tremendous potential of genomics and other recent advances in science, the role of science to improve reliable DNA extraction methods is more relevant than ever before. The ideal process for genomic DNA extraction demands high quantities of pure, integral and intact genomic DNA (gDNA from the sample with minimal co-extraction of inhibitors of downstream processes. Here, we report the development of a very rapid, less-hazardous, and high throughput protocol for extracting of high quality DNA from blood samples. Methods: Dried, clotted and ethylene diamine tetra-acetic acid (EDTA treated fresh and frozen blood samples were extracted using this method in which the quality and integrity of the extracted DNA were corroborated by agarose gel electrophoresis, PCR reaction and DNA digestion using restricted enzyme. The UV spectrophotometric and gel electrophoresis analysis resulted in high A260/A280 ratio (>1.8 with high intactness of DNA. Results: PCR and DNA digestion experiments indicated that the final solutions of extracted DNA contained no inhibitory substances, which confirms that the isolated DNA is of good quality. Conclusion: The high quality and quantity of current method, no enzymatic processing and accordingly its low cost, make it appropriate for DNA extraction not only from human but also from animal blood samples in any molecular biology labs.

  3. Detection of Candida albicans DNA from blood samples using a novel electrochemical assay.

    Science.gov (United States)

    Muir, Alastair; Forrest, Gordon; Clarkson, John; Wheals, Alan

    2011-04-01

    The genus Candida contains a number of yeast species which are opportunistic pathogens and are associated with life-threatening infections in immunocompromised individuals. Provision of appropriate therapy relies on the rapid identification of the infecting species, and existing methods of identifying Candida species in clinical samples are time and resource intensive and are not always specific enough to differentiate between drug-susceptible and drug-resistant species. We have previously developed a system for the rapid detection of yeast pathogens in clinical samples using PCR followed by hybridization with a suite of five species-specific, electrochemically labelled DNA probes. The limit of detection of the assay was shown to be 37 fg (∼1 genome) per reaction using extracted genomic DNA. We carried out a study to test the limit of detection of one of the probes, CA PR3, using blood samples from a healthy donor that were spiked with genomic DNA or with C. albicans cells. Our results demonstrated a limit of detection of 37 fg (ml blood)(-1) (∼1 genome ml(-1)) using extracted DNA or 10 c.f.u. (ml blood)(-1) using C. albicans cells, indicating that the assay is capable of detecting C. albicans nucleic acid at levels that are encountered in clinical samples.

  4. Rapid diagnosis of candidaemia by real-time PCR detection of Candida DNA in blood samples.

    Science.gov (United States)

    Wellinghausen, Nele; Siegel, Dunja; Winter, Juliane; Gebert, Susanne

    2009-08-01

    This study prospectively evaluated an 18S rRNA gene-targeted real-time PCR approach in comparison with standard blood culture (BC) diagnostics for rapid diagnosis of candidaemia in a large study population of 384 patients, including 902 whole blood samples from 468 infectious episodes (IEs) of 329 adults and 55 children with haematological malignancies and various forms of immunodeficiency, and intensive care unit patients. Seven out of eight BC-proven cases (87.5 %) of candidaemia and seven out of twelve BC-positive samples (58.3 %) were positive by the Candida-specific PCR. A positive PCR result was also obtained for 28/460 BC-negative samples from IEs, including 8 patients with culture-confirmed Candida infection at primary sterile body sites. Of the PCR-positive, culture-negative patients, more than 50 % received systemic antifungal therapy. In 432/460 BC-negative IEs, the Candida specific-PCR was negative, resulting in a negative predictive value of 99.8 %. In conclusion, the Candida specific-PCR approach facilitates rapid detection of Candida DNA in blood samples of patients at risk of candidaemia within a few hours. Although standard BC diagnostics appear to remain indispensable for the detection of all cases of candidaemia, this PCR assay allowed the detection of candidaemia at a mean of 3 days earlier than BC diagnostics. Thus, it enables earlier antifungal therapy for patients with suspected candidaemia and may prevent further complications.

  5. Blood Sampling Seasonality as an Important Preanalytical Factor for Assessment of Vitamin D Status.

    Science.gov (United States)

    Bonelli, Patrizia; Buonocore, Ruggero; Aloe, Rosalia; Lippi, Giuseppe

    2016-04-01

    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. 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. 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. 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.

  6. Hematology and serum chemistry of the island spotted skunk on Santa Cruz Island.

    Science.gov (United States)

    Crooks, Kevin R; Garcelon, D K; Scott, Cheryl A; Wilcox, Jeffery T; Timm, Steven F; Van Vuren, Dirk H

    2003-04-01

    We determined serum biochemistry and hematologic values for island spotted skunks (Spilogale gracilis amphiala) on Santa Cruz Island (California, USA). Samples were collected from island spotted skunks chemically restrained with ketamine hydrochloride and acepromazine in August 1999 (dry season) and from skunks manually restrained in August 2000 (dry season) and January 2001 (wet season). One parameter, glucose, significantly differed with season, with higher levels during the wet season. Serum chemistry and hematologic profiles suggest that method of restraint (manual or chemical), as well as other methodologic details, may influence blood characteristics in the island spotted skunk.

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

    Science.gov (United States)

    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.

  8. [Guidelines for electronic-data-processing-controlled serial diagnosis of donor blood samples].

    Science.gov (United States)

    Fischer-Fröhlich, C L; Stoll, T; Hanfland, P

    1990-01-01

    Increasing performance figures and the necessity to save expenses oblige transfusion services to automatize their donors' laboratory examination. Sufficient hard- and software for sample distribution and processing is now available. Following aspects should be regarded when switching to automatic serial screening: The identity of blood-donor, donation and laboratory result will be achieved by machine readable labeling and on-line communication between working-stations and central administration. Flexibility: Easy automatic selective laboratory screening will be possible using special barcodes including sample identification and working orders. A modular hardware concept with easily accessible programming control allows it to implement new devices or methods. Ergonomy: Automatic sample processing including selective screening and simultaneous operating robotic sample processors increase working quality, sample output and time benefits. Economy: Improved working conditions will result in saving reagents and compensating staff limitations.

  9. Detection of tumor-associated cells in cryopreserved peripheral blood mononuclear cell samples for retrospective analysis.

    Science.gov (United States)

    Zhu, Peixuan; Stanton, Melissa L; Castle, Erik P; Joseph, Richard W; Adams, Daniel L; Li, Shuhong; Amstutz, Platte; Tang, Cha-Mei; Ho, Thai H

    2016-07-02

    Cryopreserved peripheral blood mononuclear cells (PBMCs) are commonly collected in biobanks. However, little data exist regarding the preservation of tumor-associated cells in cryopreserved collections. The objective of this study was to determine the feasibility of using the CellSieve™ microfiltration assay for the isolation of circulating tumor cells (CTCs) and circulating cancer-associated macrophage-like cells (CAMLs) from cryopreserved PBMC samples. Blood samples spiked with breast (MCF-7), prostate (PC-3), and renal (786-O) cancer cell lines were used to establish analytical accuracy, efficiency, and reproducibility after cryopreservation. The spiked samples were processed through Ficoll separation, and cryopreservation was followed by thawing and microfiltration. MCF-7 cells were successfully retrieved with recovery efficiencies of 90.5 % without cryopreservation and 87.8 and 89.0 %, respectively, on day 7 and day 66 following cryopreservation. The corresponding recovery efficiencies of PC-3 cells were 83.3 % without cryopreservation and 85.3 and 84.7 %, respectively, after cryopreservation. Recovery efficiencies of 786-O cells were 92.7 % without cryopreservation, and 82.7 and 81.3 %, respectively, after cryopreservation. The recovered cells retained the morphologic characteristics and immunohistochemical markers that had been observed before freezing. The protocols were further validated by quantitation of CAMLs in blood samples from two patients with renal cell carcinoma (RCC). The recovery rates of CTCs and CAMLs from cryopreserved samples were not statistically significant different (P > 0.05) from matched fresh samples. To our knowledge, this is the first report that CAMLs could be cryopreserved and analyzed after thawing with microfiltration technology. The application of microfiltration technology to cryopreserved samples will enable much greater retrospective study of cancer patients in relation to long-term outcomes.

  10. Potentiating day-old blood samples for detection of interferon-gamma responses following infection with Mycobacterium avium subsp. paratuberculosis

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Nielsen, Søren Saxmose; Jungersen, Gregers

    time interval from blood sampling to culture. The objective of the study was to assess options for use of day-old blood samples for early-stage diagnosis of MAP infections. Bovine interleukin 12 (IL-12) can induce, and IL-10 reduce, IFN-γ production. Therefore, addition of IL-12 and anti-IL-10 could...... result in production of IFN-γ in samples previously exposed to MAP antigens. Whole blood samples were collected from heifers in a Danish dairy herd known to be infected with MAP. The samples were collected on three sample dates, and on each date the blood samples were stimulated with PPDj and recombinant......The interferon gamma (IFN-γ) test measuring specific cell-mediated immune responses in whole blood can be used for diagnosis at an early stage of Mycobacterium avium subsp. paratuberculosis (MAP) infection. A major obstacle for the practical use of IFN-γ testing is the recommended maximum 8 hour...

  11. Performance evaluation of continuous blood sampling system for PET study. Comparison of three detector-systems

    CERN Document Server

    Matsumoto, K; Sakamoto, S; Senda, M; Yamamoto, S; Tarutani, K; Minato, K

    2002-01-01

    To measure cerebral blood flow with sup 1 sup 5 O PET, it is necessary to measure the time course of arterial blood radioactivity. We examined the performance of three different types of continuous blood sampling system. Three kinds of continuous blood sampling system were used: a plastic scintillator-based beta detector (conventional beta detector (BETA)), a bismuth germinate (BGO)-based coincidence gamma detector (Pico-count flow-through detector (COINC)) and a Phoswich detector (PD) composed by a combination of plastic scintillator and BGO scintillator. Performance of these systems was evaluated for absolute sensitivity, count rate characteristic, sensitivity to background gamnra photons, and reproducibility for nylon tube geometry. The absolute sensitivity of the PD was 0.21 cps/Bq for sup 6 sup 8 Ga positrons at the center of the detector. This was approximately three times higher than BETA, two times higher than COINC. The value measured with BETA was stable, even when background radioactivity was incre...

  12. Physiological and Pathological Impact of Blood Sampling by Retro-Bulbar Sinus Puncture and Facial Vein Phlebotomy in Laboratory Mice

    DEFF Research Database (Denmark)

    Teilmann, Anne Charlotte; Nygaard Madsen, Andreas; Holst, Birgitte

    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...... 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....

  13. Wuchereria bancrofti in Tanzania: microfilarial periodicity and effect of blood sampling time on microfilarial intensities

    DEFF Research Database (Denmark)

    Simonsen, Poul Erik; Niemann, L.; Meyrowitsch, Dan Wolf

    1997-01-01

    The circadian periodicity of Wuchereria bancrofti microfilarial (mf) intensities in peripheral blood was analysed in a group of infected individuals from an endemic community in north-eastern Tanzania. The mf density was quantified at two-hourly intervals for 24 hours. A clear nocturnal periodic...... pattern was observed. Mathematical analysis of the data indicated a peak at 0152 h and a periodicity index of 117.5. A periodicity equation was developed describing the average relation between mf intensity and hour of the day for the study area. Based on the observed periodicity pattern, the effect...... of blood sampling before peak time is discussed, and the importance of taking sampling time into consideration when analysing data from epidemiological studies is emphasized. A simple method is devised which can be used to adjust for the influence of time on mf intensities, in studies where accurate...

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

    Science.gov (United States)

    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.

  15. Measuring Blood Glucose Concentrations in Photometric Glucometers Requiring Very Small Sample Volumes.

    Science.gov (United States)

    Demitri, Nevine; Zoubir, Abdelhak M

    2017-01-01

    Glucometers present an important self-monitoring tool for diabetes patients and, therefore, must exhibit high accuracy as well as good usability features. Based on an invasive photometric measurement principle that drastically reduces the volume of the blood sample needed from the patient, we present a framework that is capable of dealing with small blood samples, while maintaining the required accuracy. The framework consists of two major parts: 1) image segmentation; and 2) convergence detection. Step 1 is based on iterative mode-seeking methods to estimate the intensity value of the region of interest. We present several variations of these methods and give theoretical proofs of their convergence. Our approach is able to deal with changes in the number and position of clusters without any prior knowledge. Furthermore, we propose a method based on sparse approximation to decrease the computational load, while maintaining accuracy. Step 2 is achieved by employing temporal tracking and prediction, herewith decreasing the measurement time, and, thus, improving usability. Our framework is tested on several real datasets with different characteristics. We show that we are able to estimate the underlying glucose concentration from much smaller blood samples than is currently state of the art with sufficient accuracy according to the most recent ISO standards and reduce measurement time significantly compared to state-of-the-art methods.

  16. Stability of HE4 and CA125 in blood samples from patients diagnosed with ovarian cancer

    DEFF Research Database (Denmark)

    Sandhu, Noreen; Karlsen, Mona A; Høgdall, Claus

    2014-01-01

    OBJECTIVE: To investigate the influence of handling and storage on HE4 and CA125 serum and EDTA plasma levels to clarify any important consequences for a clinical setting. METHODS: Blood samples from 13 ovarian cancer (OC) patients were collected and allowed to clot or sediment for up to 72 hours.......024). No significant difference between CA125 serum and plasma levels were found (p = 0.46). Serum and EDTA plasma samples were stable during the eight cycles of freezing and thawing (CA125: all p > 0.2; HE4: all p > 0.5). CONCLUSION: No systematic difference could be demonstrated for HE4. CA125 is not dependent...

  17. Heel blood sampling in European neonatal intensive care units: compliance with pain management guidelines

    DEFF Research Database (Denmark)

    Losacco, Valentina; Cuttini, Marina; Greisen, Gorm

    2011-01-01

    Objective To describe the use of heel blood sampling and non-pharmacological analgesia in a large representative sample of neonatal intensive care units (NICUs) in eight European countries, and compare their self-reported practices with evidence-based recommendations. Methods Information on use...... and France were the most likely, and Belgium and Spain the least likely to employ recommended combinations of evidence-based pain management measures. Conclusions Heel puncture is a common procedure in preterm neonates, but pain appears inadequately treated in many units and countries. Better compliance...

  18. Colorimetric assay for on-the-spot alcoholic strength sensing in spirit samples based on dual-responsive lanthanide coordination polymer particles with ratiometric fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Deng, Jingjing, E-mail: jjdeng@des.ecnu.edu.cn [School of Ecological and Environmental Sciences, East China Normal University, 500 Dongchuan Road, Shanghai 200241 (China); Shi, Guoyue [Department of Chemistry, East China Normal University, 500 Dongchuan Road, Shanghai 200241 (China); Zhou, Tianshu, E-mail: tszhou@des.ecnu.edu.cn [School of Ecological and Environmental Sciences, East China Normal University, 500 Dongchuan Road, Shanghai 200241 (China)

    2016-10-26

    This study demonstrates a new strategy for colorimetric detection of alcoholic strength (AS) in spirit samples based on dual-responsive lanthanide infinite coordination polymer (Ln-ICP) particles with ratiometric fluorescence. The ICP used in this study are composed of two components: one is the supramolecular Ln-ICP network formed by the coordination between the ligand 2,2’-thiodiacetic acid (TDA) and central metal ion Eu{sup 3+}; and the other is a fluorescent dye, i.e., coumarin 343 (C343), both as the cofactor ligand and as the sensitizer, doped into the Ln-ICP network through self-adaptive chemistry. Upon being excited at 300 nm, the red fluorescence of Ln-ICP network itself at 617 nm is highly enhanced due to the concomitant energy transfer from C343 to Eu{sup 3+}, while the fluorescence of C343 at 495 nm is supressed. In pure ethanol solvent, the as-formed C343@Eu-TDA is well dispersed and quite stable. However, the addition of water into ethanolic dispersion of C343@Eu-TDA destructs Eu-TDA network structure, resulting in the release of C343 from ICP network into the solvent. Consequently, the fluorescence of Eu-TDA turns off and the fluorescence of C343 turns on, leading to the fluorescent color change of the dispersion from red to blue, which constitutes a new mechanism for colorimetric sensing of AS in commercial spirit samples. With the method developed here, we could clearly distinguish the AS of different spirit samples within a wide linear range from 10% vol to 100% vol directly by “naked eye” with the help of UV-lamp (365 nm). This study not only offers a new method for on-the-spot visible detection of AS, but also provides a strategy for dual-responsive sensing mode by rational designing the optical properties of the Ln-ICP network and the guest, respectively. - Highlights: • Dual responsive lanthanide coordination polymer particles C343@Eu-TDA were synthesized. • The guest molecular coumarin 343 sensitized the luminescence of Eu

  19. Blood gas sample spiking with total parenteral nutrition, lipid emulsion, and concentrated dextrose solutions as a model for predicting sample contamination based on glucose result.

    Science.gov (United States)

    Jara-Aguirre, Jose C; Smeets, Steven W; Wockenfus, Amy M; Karon, Brad S

    2018-03-16

    Evaluate the effects of blood gas sample contamination with total parenteral nutrition (TPN)/lipid emulsion and dextrose 50% (D50) solutions on blood gas and electrolyte measurement; and determine whether glucose concentration can predict blood gas sample contamination with TPN/lipid emulsion or D50. Residual lithium heparin arterial blood gas samples were spiked with TPN/lipid emulsion (0 to 15%) and D50 solutions (0 to 2.5%). Blood gas (pH, pCO2, pO2), electrolytes (Na+, K+ ionized calcium) and hemoglobin were measured with a Radiometer ABL90. Glucose concentration was measured in separated plasma by Roche Cobas c501. Chart review of neonatal blood gas results with glucose >300 mg/dL (>16.65 mmol/L) over a seven month period was performed to determine whether repeat (within 4 h) blood gas results suggested pre-analytical errors in blood gas results. Results were used to determine whether a glucose threshold could predict contamination resulting in blood gas and electrolyte results with greater than laboratory-defined allowable error. Samples spiked with 5% or more TPN/lipid emulsion solution or 1% D50 showed glucose concentration >500 mg/dL (>27.75 mmol/L) and produced blood gas (pH, pO 2 , pCO 2 ) results with greater than laboratory-defined allowable error. TPN/lipid emulsion, but not D50, produced greater than allowable error in electrolyte (Na + ,K + ,Ca ++ ,Hb) results at these concentrations. Based on chart review of 144 neonatal blood gas results with glucose >250 mg/dL received over seven months, four of ten neonatal intensive care unit (NICU) patients with glucose results >500 mg/dL and repeat blood gas results within 4 h had results highly suggestive of pre-analytical error. Only 3 of 36 NICU patients with glucose results 300-500 mg/dL and repeat blood gas results within 4 h had clear pre-analytical errors in blood gas results. Glucose concentration can be used as an indicator of significant blood sample contamination with either TPN

  20. TCRgamma gene rearrangement analysis in skin samples and peripheral blood of mycosis fungoides patients.

    Science.gov (United States)

    Kandolf Sekulović, L; Cikota, B; Stojadinović, O; Basanović, J; Skiljević, D; Medenica, Lj; Pavlović, M; Magić, Z

    2007-12-01

    Diagnosing mycosis fungoides (MF) can be challenging in the early stage of the disease because histopathological features may simulate a variety of benign inflammatory skin diseases. Assessment of T-cell clonality was found to be useful in diagnosis and follow-up of patients. In this study, PCR-based TCRgamma gene rearrangement analysis was performed in skin and peripheral blood samples of patients with MF treated at the two largest referral centers in Serbia, and the results obtained were correlated with clinical and follow-up data. Skin and peripheral blood samples were obtained with informed consent from 37 patients treated at the Department of Dermatology of the Military Medical Academy and the Medical Center of Serbia from 2001 to 2006. The median time of follow-up was 4 years. Multiplex PCR was used for TCRgamma gene rearrangement analysis in skin and peripheral blood samples. Clonality results were correlated with the clinical data and disease course data. Monoclonality was detected in skin samples of 30/37 patients (81%), in 2/5 patients with large-plaque parapsoriasis (LPP), in 28/32 (88%) patients with histologically proven MF, and in 1/16 (6%) patients with benign inflammatory dermatoses. A monoclonal pattern in both skin and peripheral blood was detected in 7/16 (44%) patients in the late stage of the disease, and in 1/7 (14%) patients in the early stage of the disease. A dominant clone was found in both skin and peripheral blood in 1/4 patients in remission, 2/5 with a stable disease, and 4/9 (44%) with disease progression. TCR-gamma gene rearrangement analysis can be regarded as a useful adjunct to diagnosis of epidermotropic lymphoproliferative disorders. The presence of a dominant clone in both the skin and peripheral blood was more frequently detected in late stages and in patients with disease progression, confirming the usefulness of clonality detection by TCR-gamma gene rearrangement analysis in follow-up of patients with primary cutaneous T

  1. Dendritic cells in blood and urine samples from bladder cancer patients undergoing BCG immunotherapy

    Directory of Open Access Journals (Sweden)

    Raffaella Rossi

    2013-12-01

    Full Text Available Objectives: Immunotherapy with BCG (Bacille Calmette-Guérin after transurethral resection of the bladder tumor represents a highly effective primary treatment for intermediate and high-risk superficial bladder cancer. The effectiveness of this therapy has been documented, but its mechanism of action is not clear yet. In the present study, we investigated the changes of dendritic cells (DC numbers in peripheral blood and urine of patients with superficial bladder cancer undergoing BCG intravescical therapy Material and method: We have enumerated plasmacytoid and myeloid DCs in the peripheral blood and in the urine of patients with bladder cancer in order to clarify the role of these cells in the evolution of the disease and the effect of therapy. DCs in blood and urine samples were assessed using the single-platform TruCOUNT assay with monoclonal antibodies. The study population included 37 healthy donors and 13 patients with diagnosis of primitive superficial bladder cancer. Results: At the time of diagnosis a reduction of blood DCs was found in patients as opposed to healthy donors, while DCs were not found in the urine in the same way as in healthy subjects. Six of these patients were followed before and after weekly and monthly instillations of BCG. In the peripheral blood, we observed an immunological recovery of DCs from the third weekly instillation up to the sixth. In the urine of patients, we didn’t find mDCs or pDCs at T0, but we found a statistically significant change from the third instillation up to the sixth. On the contrary, we didn’t find mDCs in urine during monthly instillation. Conclusions: DC Count could be used in the monitoring of patients undergoing BCG therapy. Immunological restoration of mDC numbers in peripheral blood and the efflux in urine could be important for confirming the effectiveness of BCG instillation.

  2. Classical and additional antiphospholipid antibodies in blood samples of ischemic stroke patients and healthy controls.

    Science.gov (United States)

    Carmel-Neiderman, Narin-Nard; Tanne, David; Goren, Idan; Rotman-Pikielny, Pnina; Levy, Yair

    2017-04-01

    Classical antiphospholipid antibodies (aPLa) are found in 6-25% of blood samples from stroke patients. The frequency of novel aPLa antibodies in blood samples of CVA patients is not known. Enzyme-linked immunosorbent assays (ELISA) were performed on blood samples from 209 CVA patients (170 samples were obtained during the acute phase and 39 samples were from patients with complete carotid stenosis) and compared to 54 healthy controls. Subjects were tested for the presence of the classical aPL antibodies anticardiolipin (aCL) and anti-beta2-glycoprotein (aβ2gI), in addition to antiphosphatidylethanolamine (aPE), anti-phosphatidylserine (aPS), and Annexin V. All antibodies were tested for both IgM and IgG subclasses. Numeric analysis of the antibody titer levels (μ/ml) revealed a significantly higher subclinical titer by two standard deviations of many aPL autoantibodies among CVA patients (Pv < 0.05). However, according to the kit manufacturer's cutoff value, no positive antibodies were found except a trend toward higher percentage of positive aPS IgG titer in the CVA group compared to controls (6.2 vs. %0; P = 0.077). According to the manufacturer's cutoff, significantly higher levels of positive antibodies were not found among stroke patients. However, the absolute ELISA values of stroke patients were significantly higher. These results suggest that lower cutoff values than those used for APS diagnosis should be used for risk stratification of CVA among healthy individuals.

  3. Sensitivity of PCR assays for murine gammaretroviruses and mouse contamination in human blood samples.

    Directory of Open Access Journals (Sweden)

    Li Ling Lee

    Full Text Available 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.

  4. Leukocyte count affects expression of reference genes in canine whole blood samples

    Directory of Open Access Journals (Sweden)

    Dekker Aldo

    2011-02-01

    Full Text Available Abstract 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.

  5. Frequency and antimicrobial susceptibility of acinetobacter species isolated from blood samples of paediatric patients

    International Nuclear Information System (INIS)

    Javed, A.; Zafar, A.; Ejaz, H.; Zubair, M.

    2012-01-01

    Objective: Acinetobacter species is a major nosocomial pathogen causing serious infections in immuno-compromised and hospitalized patients. The aim of this study was to determine the frequency and antimicrobial susceptibility pattern of Acinetobacter species in blood samples of paediatric patients. Methodology: This cross sectional observational study was conducted during January to October, 2011 at The Children's Hospital and Institute of Child Health, Lahore. A total number of 12,032 blood samples were analysed during the study period. Acinetobacter species were Bauer disc diffusion method. Results: The blood cultures showed growth in 1,141 cultures out of which 46 (4.0%) were Acinetobacter species. The gender distribution of Acinetobacter species was 29 (63.0%) in males and 17 (37.0%) in females. A good antimicrobial susceptibility pattern of Acinetobacter species was seen with sulbactam-cefoperazone (93.0%), imepenem and meropenem (82.6% (30.4%) was poor. Conclusion: The results of the present study shows high rate of resistance of Acinetobacter species with cephalosporins in nosocomial infections. The sulbactam-cefoperazone, carbapenems and piperacillin-tazobactam showed effective antimicrobial susceptibility against Acinetobacter species. (author)

  6. Application of Atomic Dielectric Resonance Spectroscopy for the screening of blood samples from patients with clinical variant and sporadic CJD

    Directory of Open Access Journals (Sweden)

    Ironside James W

    2007-08-01

    Full Text Available Abstract 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.

  7. Application of Atomic Dielectric Resonance Spectroscopy for the screening of blood samples from patients with clinical variant and sporadic CJD

    Science.gov (United States)

    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

  8. Concentrations of cadmium, lead, and zinc in fish from mining-influenced waters of northeastern Oklahoma: Sampling of blood, carcass, and liver for aquatic biomonitoring

    Science.gov (United States)

    Brumbaugh, W.G.; Schmitt, C.J.; May, T.W.

    2005-01-01

    The Tri-States Mining District (TSMD) of Missouri (MO), Kansas (KS), and Oklahoma (OK), USA, was mined for lead (Pb) and zinc (Zn) for more than a century. Mining ceased more than 30 years ago, but wastes remain widely distributed in the region, and there is evidence of surface- and groundwater contamination in the Spring River-Neosho River (SR-NR) system of northeastern OK. In October 2001, we collected a total of 74 fish from six locations in the SR-NR system that included common carp (Cyprinus carpio), channel- and flathead catfish (Ictalurus punctatus and Pylodictis olivaris), largemouth- and spotted bass (Micropterus salmoides and Micropterus punctulatus), and white crappie (Pomoxis annularis). We obtained additional fish from locations in MO that included three reference sites and one site that served as a "positive control" (heavily contaminated by Pb). Blood, carcass (headed, eviscerated, and scaled) and liver (carp only) samples were analyzed for cadmium (Cd), Pb, and Zn. Our objectives were to assess the degree to which fish from the OK portion of the SR-NR system are contaminated by these elements and to evaluate fish blood sampling for biomonitoring. Concentrations of Cd and Pb in carp and catfish from OK sites were elevated and Pb concentrations of some approached those of the highly contaminated site in MO, but concentrations in bass and crappie were relatively low. For Zn, correlations were weak among concentrations in the three tissues and none of the samples appeared to reflect site contamination. Variability was high for Cd in all three tissues of carp; differences between sites were statistically significant (p < 0.05) only for blood even though mean liver concentrations were at least 100-fold greater than those in blood. Blood concentrations of Cd and Pb were positively correlated (r 2 = 0.49 to 0.84) with the concentration of the same element in carp and catfish carcasses or in carp livers, and the corresponding multiple regression models were

  9. Detection of HTLV-I and -II in Scottish blood donor samples and archive donations.

    Science.gov (United States)

    Davidson, F; Lycett, C; Jarvis, L M; Kerr, D; Lumley, S; Petrik, J; Dow, B C

    2006-10-01

    Positive samples identified during routine serological screening for HCV (hepatitis C virus), HBV (hepatitis B virus) and HIV (human immunodeficiency virus) are confirmed by nucleic acid testing in the SNBTS (Scottish National Blood Transfusion Service) PCR Reference laboratory. Serological screening for HTLV-I (human T-cell lymphotropic virus type I) and -II was implemented in Scotland in November 2002, at which time a PCR assay was not available for confirmation. Our aim was to develop a real-time PCR assay that could be used for the confirmation of samples showing HTLV-I serological positive or indeterminate reactivity and to investigate whether a serologically silent carrier status exists ('Tax' only) in the Scottish donor population. A real-time HTLV PCR was devised using a lymphoblastoid cell line which has HTLV-I sequence integrated in the genome (C8166 cells). These were spiked into peripheral blood mononuclear cells. The assay was evaluated on archived serologically confirmed HTLV-positive samples and new positives identified since implementation of screening. HTLV-I and -II were detected in cells and plasma from stored donations and a serological positive donation identified in routine screening. HTLV DNA can also be amplified from the plasma obtained from plasma preparation tubes. There was no evidence of a carrier status ('Tax' only) in 100 serologically negative blood donors tested. The PCR assay developed is reliable and sensitive, capable of identifying one copy of HTLV-I. The HTLV PCR is a useful addition for HTLV confirmation, especially in serologically indeterminate samples and for look-back studies. HTLV PCR confirmation will provide additional useful information for donor medical staff for counselling donors.

  10. Leakage of Oxygen from Blood and Water Samples Stored in Plastic and Glass Syringes

    Science.gov (United States)

    Scott, Peter V.; Horton, J. N.; Mapleson, W. W.

    1971-01-01

    Theory and experiment showed that samples of blood and water stored in 2-ml and 5-ml syringes made of polypropylene, polystyrene, or S.A.N. co-polymer exchanged oxygen with their surroundings. In the first hour the exchange was due mainly to equilibration with the plastic of the syringe and only in small degree to permeation through the plastic. With high initial tension or with blood of low haemoglobin concentration the exchange can result in errors in Po2 of up to 6% in two minutes and 16% in 30 to 60 minutes. With all-glass syringes the exchange was much slower but, even so, after 24 hours was important in all but a few of 18 interchangeable glass syringes. Therefore unless analysis can be started immediately all-glass syringes are to be preferred, and for prolonged storage even these should be selected. PMID:5565518

  11. Role of plasmin on the double antibody radioimmunoassay of carcinoembryonic antigen in human blood samples

    International Nuclear Information System (INIS)

    Das, S.; Das, B.R.

    1976-01-01

    Double antibody radioimmunoassay (RIA) of carcinoembryonic antigen (CEA) on a series of freshly drawn out concurrent plasma and serum samples of normal human blood donors showed that the serum CEA values were invariably higher than the corresponding plasma CEA values. Extraneous addition of fibrinogen brought down the serum--CEA level to a value comparable to or less than the corresponding plasma value. The effect of certain factors associated with blood clotting, particularly Ca ++ , fibrinogen, and the fibrinolytic enzyme plasmin, was investigated. Ca ++ was shown to play no role whereas the effect of fibrinogen was shown to be indirect in that it served as a specific substrate for plasmin, thereby preventing the plasmin degradation of the primary antibody used in the RIA. The finding stresses the role of enzymes like plasmin in double antibody RIA in general and may explain some of the anomalous results obtained when testing biologic material containing plasmin-like substances

  12. Validation of curve-fitting method for blood retention of {sup 99m}Tc-GSA. Comparison with blood sampling method

    Energy Technology Data Exchange (ETDEWEB)

    Ha-Kawa, Sang Kil; Suga, Yutaka; Kouda, Katsuyasu; Ikeda, Koshi; Tanaka, Yoshimasa [Kansai Medical Univ., Moriguchi, Osaka (Japan)

    1997-02-01

    We investigated a curve-fitting method for the rate of blood retention of {sup 99m}Tc-galactosyl serum albumin (GSA) as a substitute for the blood sampling method. Seven healthy volunteers and 27 patients with liver disease underwent {sup 99m}Tc-GSA scanning. After normalization of the y-intercept as 100 percent, a biexponential regression curve for the precordial time-activity curve provided the percent injected dose (%ID) of {sup 99m}Tc-GSA in the blood without blood sampling. The discrepancy between %ID obtained by the curve-fitting method and that by the multiple blood samples was minimal in normal volunteers 3.1{+-}2.1% (mean{+-}standard deviation, n=77 sampling). Slightly greater discrepancy was observed in patients with liver disease (7.5{+-}6.1%, n=135 sampling). The %ID at 15 min after injection obtained from the fitted curve was significantly greater in patients with liver cirrhosis than in the controls (53.2{+-}11.6%, n=13; vs. 31.9{+-}2.8%, n=7, p<0.0001). There was a highly linear correlation between the %IDs of {sup 99m}Tc-GSA and the plasma retention rate for indocyanine green (r=-0.869, p<0.0001, n=27). These results indicate that the curve-fitting method provides an accurate %ID of {sup 99m}Tc-GSA and could be a substitute for the blood sampling method. (author)

  13. Less invasive blood sampling in the animal laboratory: clinical chemistry and haematology of blood obtained by the Triatominae bug Dipetalogaster maximus.

    Science.gov (United States)

    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.

  14. Prospective comparison of a PCR assay and a microbiological culture technique for identification of pathogens from blood and non-blood samples in septic patients.

    Science.gov (United States)

    Plettig, Runa; Nowak, Andreas; Balau, Veronika; Hahnenkamp, Klaus; Usichenko, Taras

    2015-01-01

    Molecular amplification techniques are suggested to be a useful adjunct in early detection of pathogens in septic patients. The aim was to study the feasibility of a polymerase chain reaction (PCR) assay compared to the standard microbiological culture (MC) technique in identification of pathogenic microorganisms from blood and non-blood samples in septic patients. Samples for pathogen identification were taken during febrile septic episodes (SE) in 54 patients with sepsis and analyzed using both MC and PCR. Semi-automated multiplex PCR, provided by Philips Medical Systems, was able to detect nine different pathogens. The accuracy of pathogen identification using PCR vs. MC as well as the time-saving effect of PCR on the potential decision-making process for antimicrobial therapy was evaluated. In a total of 258 samples taken during 87 SE, both methods yielded more pathogens from the non-blood than blood samples (87 % vs. 45 %; p = 0.002). PCR identified more pathogens than MC in the blood samples (98 vs. 21; p technique. In the non-blood samples, PCR was comparable to that of MC.

  15. Quantification and Optimization of Candida albicans DNA in Blood Samples Using Real- Time PCR

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    Mojtaba Nabili

    2013-10-01

    Full Text Available Background: Candida albicans (C. albicans is a major cause of candidaemia in people with impaired immunity. Blood culture is a “gold standard” for candidaemia detection but is time-consuming and relatively insensitive. We established a real-time PCR assay for C. albicans detection in blood by LightCycler PCR and melting curve analysis. Methods: Five milliliter blood samples from healthy volunteers were spiked with 100-106 C. albicans cells to determine the detection limit of our method. DNA was extracted from whole blood using glass beads and the QIAamp DNA Blood Mini Kit (Qiagen, Hilden Germany. DNA from C. albicans isolates were amplified with primers and inserted into Escherichia coli (E. coli DH5α.1 cells with the TA cloning vector (Invitrogen. The plasmid was used for standardization and optimization. A quantitative PCR assay with the LightCycler amplification and detection system based on fluorescence resonance energy transfer (FRET with two different specific probes was established. To assess the precision and reproducibility of real-time PCR the intra-assay precision was determined in six consecutive assays. Results: No cross-reactivity of the hybridization probes with the DNA of non-C. albicans species or human genomic DNA was observed, which confirmed its 100% specificity. The minimum limit detected was one C. albicans cell or 100 CFU/ml (10 fg per PCR reaction. The real-time PCR efficiency rate for Candida was high (E = 1.95. Melting curve analysis of C. albicans showed a specific melting peak temperature of 65.76 °C. Conclusion: The real-time PCR assay we developed is highly specific and sufficiently sensitive to detect the fungal load for early diagnosis of invasive candidiasis.

  16. Quantification and optimization of Candida albicans DNA in blood samples using Real- Time PCR.

    Science.gov (United States)

    Nabili, Mojtaba; Ashrafi, Mohsen; Janbabaie, Ghasem; Hedayati, Mohamad Taghi; Ali-Moghaddam, Kamran; Shokohi, Tahereh

    2013-10-01

    Candida albicans (C. albicans) is a major cause of candidaemia in people with impaired immunity. Blood culture is a "gold standard" for candidaemia detection but is time-consuming and relatively insensitive. We established a real-time PCR assay for C. albicans detection in blood by LightCycler PCR and melting curve analysis. Five milliliter blood samples from healthy volunteers were spiked with 10(0)-10(6) C. albicans cells to determine the detection limit of our method. DNA was extracted from whole blood using glass beads and the QIAamp DNA Blood Mini Kit (Qiagen, Hilden Germany). DNA from C. albicans isolates were amplified with primers and inserted into Escherichia coli (E. coli) DH5α.1 cells with the TA cloning vector (Invitrogen). The plasmid was used for standardization and optimization. A quantitative PCR assay with the LightCycler amplification and detection system based on fluorescence resonance energy transfer (FRET) with two different specific probes was established. To assess the precision and reproducibility of real-time PCR the intra-assay precision was determined in six consecutive assays. No cross-reactivity of the hybridization probes with the DNA of non-C. albicans species or human genomic DNA was observed, which confirmed its 100% specificity. The minimum limit detected was one C. albicans cell or 10(0) CFU/ml (10 fg) per PCR reaction. The real-time PCR efficiency rate for Candida was high (E = 1.95). Melting curve analysis of C. albicans showed a specific melting peak temperature of 65.76 °C. The real-time PCR assay we developed is highly specific and sufficiently sensitive to detect the fungal load for early diagnosis of invasive candidiasis.

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

    Science.gov (United States)

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

    2015-01-01

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

  18. Identification of a suitable internal control for fluorescence analysis on canine peripheral blood samples.

    Science.gov (United States)

    Riondato, F; Martini, V; Poggi, A; Rota, A; Comazzi, S; Sulce, M; Bruno, B; Borrelli, A; Miniscalco, B

    2016-04-01

    Reliable detection of fluorescence intensity (FI) by flow cytometry (FC) is fundamental. FI depends on instrument settings and sample processing procedures: thus, measurements should be done using internal controls with known FI. Commercially available beads-based standards are expensive, thus reducing their usability in the veterinary practice. Cell subsets with stable mean FI (MFI) within the population have been proposed as acceptable surrogates in human medicine. In veterinary medicine, no data exist about stability of antigen expression among different subjects or upon sample storage. The aim of the present study was to evaluate MFI variability of main lymphocytes antigens among the lymphoid cells within each subject, among different subjects, and upon 24-h storage, in order to identify the antigen most suitable as stable internal control in MFI analyses. Peripheral blood samples from 18 healthy dogs were analysed by FC within 3h from sampling to assess the expression of CD3, CD5, CD4, CD8, CD21 and cyCD79b using conjugated monoclonal antibodies. Analyses were restricted to the lymphoid population. Fluorescent microbeads were added to each tube, and antigen MFI was calculated as Relative Fluorescence Intensity RFI (CD/beads). Fluorescence histogram CV (fhCV) for each CD was regarded as an index of the variability of expression among lymphocytes within each subject (cell-to-cell variability); whereas the CV of RFI was regarded as an index of inter-subjects variability (dog-to-dog variability). In 11 cases, FC analyses were repeated after 24h storage at 4°C and RFI and CVs of fresh and stored samples were compared to assess variability linked to storage. CD4 was identified as the best antigen to be used as an internal control for MFI analyses in canine peripheral blood samples because of low cell-to-cell and dog-to-dog variability, and optimal stability upon 24-h storage. Blood samples from a second group of 21 healthy dogs were labelled only with CD4, in order

  19. Detection of Babesia DNA in blood and spleen samples from Eurasian badgers (Meles meles) in Scotland.

    Science.gov (United States)

    Bartley, Paul M; Wilson, Cari; Innes, Elisabeth A; Katzer, Frank

    2017-08-01

    Babesia are intraerythrocytic parasites of importance worldwide within the fields of human and veterinary medicine, as some Babesia sp., including Babesia microti are potentially zoonotic and can cause fatal disease in both humans and animals. The aims of this study were to use a nested PCR (amplifying the 18S rRNA gene) to determine the presence and species of Babesia parasite DNA found in blood (n = 47) and spleen (n = 47) samples collected from Eurasian badgers (Meles meles) in Scotland. The results showed 28/47 (59·6%) blood and 14/47 (29·8%) spleen samples tested positive for the presence of Babesia DNA. Initial sequence analysis of the Babesia DNA identified three distinct sequence types (submitted to GenBank KX528553, KX528554 and KX528555), which demonstrated ⩾99% identity to Babesia sp. parasites previously identified in badgers in Spain (KT223484 and KT223485). Phylogenetic analysis showed that the three isolates are closely related to Babesia annae, B. microti and other Piroplasmida species found in wildlife. Further sequence analysis of the samples demonstrated that the badgers were routinely infected with more than one parasite isolate and there was also evidence of genetic recombination between the Babesia parasite isolates (submitted to GenBank KY250472 - KY250477).

  20. Trace elements in blood samples of workers in Atbara railways foundry

    International Nuclear Information System (INIS)

    Mustafa, W. M.

    2013-09-01

    This study was conducted to determine trace elements and toxic substances in biological samples (blood samples) of humans. The aim of the current study was to determine the concentration of iron (Fe), copper (Cu), (Pb), lead, and zinc (Zn) in biological samples of workers employed in the industrial workshops in the River Nile state to assess the potential impact of exposure to the work environmental factors. For the purpose of comparison biological samples were collected from the same group of workers exposed to the elements of the work environment and workers not exposed to the elements of the work environment. The analysis of all elements in biological samples was done by x-ray fluorescence technique (X RF). There were no statistically significant differences between the analytical results for the exposed group and non-exposed group, using the same technique. The results showed that the concentrations of the four elements copper, lead, iron, and zinc in all biological samples from workers exposed were not much higher than those not exposed, it could be argued that there was a possible link between these elements with different causes of physiological disorder. The results also showed that need for an attention for improvements in hygiene practice in the workplace and industrial ventilation.(Author)

  1. Capillary blood sampling: national recommendations on behalf of the Croatian Society of Medical Biochemistry and Laboratory Medicine.

    Science.gov (United States)

    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.

  2. Weekday variation in triglyceride concentrations in 1.8 million blood samples

    DEFF Research Database (Denmark)

    Jaskolowski, Jörn; Ritz, Christian; Sjödin, Anders Mikael

    2017-01-01

    BACKGROUND: Triglyceride (TG) concentration is used as a marker of cardio-metabolic risk. However, diurnal and possibly weekday variation exists in TG concentrations. OBJECTIVE: To investigate weekday variation in TG concentrations among 1.8 million blood samples drawn between 2008 and 2015 from...... variations in TG concentrations were recorded for out-patients between the age of 9 to 26 years, with up to 20% higher values on Mondays compared to Fridays (all PTriglyceride concentrations were highest after the weekend and gradually declined during the week. We suggest that unhealthy...

  3. 7 CFR 28.423 - Middling Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody of...

  4. Validation of a fully automated robotic setup for preparation of whole blood samples for LC-MS toxicology analysis

    DEFF Research Database (Denmark)

    Andersen, David Wederkinck; Rasmussen, Brian; Linnet, Kristian

    2012-01-01

    -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......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...... 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...

  5. Concentrations of persistent organic pollutants (POPs) in human blood samples from Mexico City, Mexico.

    Science.gov (United States)

    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 (

  6. Neonatal vitamin D status from archived dried blood spots and future risk of fractures in childhood: results from the D-tect study, a population-based case-cohort study.

    Science.gov (United States)

    Händel, Mina Nicole; Frederiksen, Peder; Cohen, Arieh; Cooper, Cyrus; Heitmann, Berit Lilienthal; Abrahamsen, Bo

    2017-07-01

    Background: Whether antenatal and neonatal vitamin D status have clinical relevance in fracture prevention has not been examined extensively, although observational studies indicate that fetal life may be a sensitive period in relation to bone growth and mineralization during childhood. Objective: We examined whether 25-hydroxyvitamin D 3 [25(OH)D 3 ] concentrations in stored neonatal dried blood spot (DBS) samples are associated with pediatric fracture risk. We hypothesized that in particular, low neonatal vitamin D status may be a risk factor for fracture incidence among children. Design: In a register-based case-cohort study design, the case group was composed of 1039 individuals who were randomly selected from a total of 82,154 individuals who were born during 1989-1999 and admitted to a Danish hospital with a fracture of the forearm, wrist, scaphoid bone, clavicle, or ankle at age 6-13 y. The subcohort was composed of 1600 individuals randomly selected from all Danish children born during 1989-1999. The neonatal 25(OH)D 3 concentrations in DBS samples were assessed by using highly sensitive chromatography-tandem mass spectrometry. Results: The mean ± SD 25(OH)D 3 concentration for all subjects was 27.7 ± 18.9 nmol/L [median (IQR): 23.5 nmol/L (13.3, 37.3 nmol/L)] and showed significant monthly variation ( P fracture than did those in the lowest quintile (adjusted OR: 0.75; 95% CI: 0.58, 0.96), but a global test did not show any significant overall association (adjusted P = 0.13). Conclusions: This study suggested that neonatal vitamin D status does not influence subsequent fracture risk in childhood. This is in accordance with studies that report no association between antenatal maternal vitamin D status and childhood fractures. Further studies are needed to examine fracture risk in relation to prenatal vitamin D status in a randomized controlled setting. © 2017 American Society for Nutrition.

  7. Thyroxine and thyrotropin radioimmunoassays using dried blood samples on filter paper for screening of neonatal hypothyroidism

    International Nuclear Information System (INIS)

    Beckers, C.; Cornette, C.; Francois, B.; Bouckaert, A.; Lechat, M.

    1977-01-01</