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Dried Blood Spots (dried + blood_spot)

Distribution by Scientific Domains

Medical Sciences	66%
Life Sciences	16%
Chemistry	16%

Selected Abstracts

A new rapid micromethod for the assay of phenobarbital from dried blood spots by LC-tandem mass spectrometry

EPILEPSIA, Issue 12 2009
Giancarlo La Marca
Summary Advantages of dried blood spot include low invasiveness, ease and low cost of sample collection, transport, and storage. We used tandem mass spectrometry (LC-MS/MS) to determine phenobarbital levels on dried blood spot specimens and compared this methodology to commercially available particle enhanced turbidimetric inhibition immunoassay (PETINIA) in plasma/serum samples. The calibration curve in matrix using D5 -phenobarbital as internal standard was linear in the phenobarbital concentration range of 1,100 mg/L (correlation coefficient 0.9996). The coefficients of variation in blood spots ranged 2.29,6.71% and the accuracy ranged 96.54,103.87%. There were no significant differences between the concentrations measured using PETINA and LC-MS/MS (both had similar precision and accuracy) however, LC-MS/MS allows at least 1.5 times higher throughput of phenobarbital analysis and additionally offers ease of sample collection which is particularly important for newborns or small infants. [source]

Human immunodeficiency virus serotyping on dried serum spots as a screening tool for the surveillance of the AIDS epidemic

JOURNAL OF MEDICAL VIROLOGY, Issue S1 2006
Francis Barin
Abstract Many studies have demonstrated the utility of the dried blood spot (DBS) or dried plasma/serum spot (DSS) method for serological and molecular diagnosis of HIV infection. Here, we report on the description of a serotyping assay performed on DSS, and its application to a national surveillance program of HIV variants. We combined serotyping assays that we developed previously to discriminate between HIV-1 and HIV-2, between HIV-1 group O and HIV-1 group M, and between B and non-B subtypes of HIV-1 group M. The assays are based on antibody binding to either the immunodominant epitope of gp41 or the V3 domain of gp120 of these various types, groups and subtypes. Therefore, a unique enzyme-linked immunosorbent assay (ELISA) format applied to serum eluted from DSS allowed the simultaneous discrimination between infections caused by HIV-1 B, HIV-1 non-B, HIV-1 group O, and HIV-2. Together, this serotyping assay and an immunoassay for recent infection were used for a virological surveillance linked to the anonymous mandatory notification of HIV infection in France. The preliminary results of this virological surveillance allowed us to obtain estimates of the prevalence of the rare variants HIV-2 and HIV-1 group O. It also allowed identification of the two first cases of M/O dual infections reported outside the endemic group O region of the western part of equatorial Africa, and showed that non-B subtypes circulate widely in France, almost 50% of new HIV diagnoses in 2003 being due to these variants. J. Med. Virol. 78:S13,S18, 2006. © 2006 Wiley-Liss, Inc. [source]

Increasing the uptake of hepatitis C virus testing among injecting drug users in specialist drug treatment and prison settings by using dried blood spots for diagnostic testing: a cluster randomized controlled trial

JOURNAL OF VIRAL HEPATITIS, Issue 4 2008
M. Hickman
Summary., The objective of this study was to assess whether introducing dried blood spot testing can increase hepatitis C virus (HCV) diagnostic testing. A cluster randomized controlled trial was conducted. Sites were matched into pairs, with one site in each pair randomly allocated to receive the intervention (training and use of dried blood spot). Data were collected from all sites for 6 months before and 6 months after the start of the intervention. The participants were 22 specialist drug clinics and six prisons in England and Wales. The main outcome measure of this study was percentage point difference in individuals tested for HCV (the difference between the percentage of patients tested 6 months after and 6 months before the introduction of dried blood spot tests). Before the trial, 8% of patients at control and intervention sites had been tested for HCV, with 16 sites testing less than 5% of their caseload. The average percentage point difference between intervention and control sites was 14.5% (95% CI 1.3,28%, paired t -test, P = 0.03); with 13 of the 14 pairs contributing to the positive effect of the intervention (Wilcoxon matched-pairs signed-rank-test, P = 0.002). The size of the difference between intervention and control sites varied considerably. The study provides preliminary supporting evidence that dried blood spot testing may increase the uptake of HCV diagnostic testing, by increasing the opportunity for patients to be offered testing. Additional trials with a larger number of sites are justified, ideally in the context of drug and treatment policies that gave clearer priority (and targets) to infection control and testing. [source]

Precursor ion scan profiles of acylcarnitines by atmospheric pressure thermal desorption chemical ionization tandem mass spectrometry

RAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 23 2008
Giuseppe Paglia
The fatty acyl esters of L-carnitine (acylcarnitines) are useful biomarkers for the diagnosis of some inborn errors of metabolism analyzed by liquid chromatography/tandem mass spectrometry. In this study the acylcarnitines were analyzed by atmospheric pressure thermal desorption chemical ionization using a commercial tandem mass spectrometer (APTDCI-MS/MS). The method is based on the precursor ion scan mode determination of underivatized acylcarnitines desorbed from samples by a hot desolvation gas flow and ionized by a corona pin discharge. During desorption/ionization step the temperature induces the degradation of acylcarnitines; nevertheless, the common fragment to all acylcarnitines [MH,59]+ is useful for analyzing their profile. APTDCI parameters, including angle of collection and incidence, gas flows and temperatures, were optimized for acylcarnitines. The experiments were performed drying 2,µL of an equimolar mixture of acylcarnitine standards on a glass slide. The specificity was evaluated by comparing product ion spectra and the precursor ion spectra of 85 m/z of acylcarnitines obtained by the APTDCI method and by electrospray ionization flow injection analysis (ESI-FIA). The method was also employed to analyze acylcarnitines extracted from a pathological dried blood spot and a control. The method enables analysis of biological samples and recognition of some acylcarnitines that are diagnostic markers of inherited metabolic diseases. The intrinsic high-throughput analysis of the ambient desorption ionization methods offers a new opportunity either for its potential application in clinical chemistry and for the expanded screening of some inborn errors of metabolism. Copyright © 2008 John Wiley & Sons, Ltd. [source]

Dried blood spot sampling in combination with LC-MS/MS for quantitative analysis of small molecules

BIOMEDICAL CHROMATOGRAPHY, Issue 1 2010
Wenkui Li
Abstract The collection of whole blood samples on paper, known as dried blood spot (DBS), dates back to the early 1960s in newborn screening for inherited metabolic disorders. DBS offers a number of advantages over conventional blood collection. As a less invasive sampling method, DBS offers simpler sample collection and storage and easier transfer, with reduced infection risk of various pathogens, and requires a smaller blood volume. To date, DBS-LC-MS/MS has emerged as an important method for quantitative analysis of small molecules. Despite the increasing popularity of DBS-LC-MS/MS, the method has its limitations in assay sensitivity due to the small sample size. Sample quality is often a concern. Systematic assessment on the potential impact of various blood sample properties on accurate quantification of analyte of interest is necessary. Whereas most analytes may be stable on DBS, unstable compounds present another challenge for DBS as enzyme inhibitors cannot be conveniently mixed during sample collection. Improvements on the chemistry of DBS card are desirable. In addition to capturing many representative DBS-LS-MS/MS applications, this review highlights some important aspects of developing and validating a rugged DBS-LC-MS/MS method for quantitative analysis of small molecules along with DBS sample collection, processing and storage. Copyright © 2009 John Wiley & Sons, Ltd. [source]

A new rapid micromethod for the assay of phenobarbital from dried blood spots by LC-tandem mass spectrometry

Molecular neonatal screening for homocystinuria in the Qatari population,

HUMAN MUTATION, Issue 6 2009
Johannes Zschocke
Abstract We report the results of molecular neonatal screening for homocystinuria (cystathionine beta-synthase deficiency) in neonates of Qatari origin, developed in conjunction with a novel biochemical screening approach. DNA was extracted from dried blood spots (DBS); the prevalent Qatari CBS gene mutation p.R336C (c.1006C>T) and a second mutation were tested with specific TaqMan assays. Over a period of 2 years we screened 12,603 neonates and identified six affected neonates homozygous for p.R336C. There were 225 heterozygous carriers for p.R336C. One additional child with homocystinuria detected through biochemical screening was homozygous for a mutation not previously identified in Qatar. Homocystinuria in the Qatari population has an incidence of 1:1,800, the highest in the world and even higher than previously estimated. Allele frequency of the mutation p.R336C is approximately 1%, displaying a significant deviation from Hardy Weinberg equilibrium. In conclusion, first-line molecular neonatal screening is technically feasible and may be developed as an option for presymptomatic identification of genetic disorders caused by specific mutations or a limited number of prevalent mutations. However, sensitivity for the diagnosis of disorders caused by various mutations is limited even in a homogeneous population such as Qatar. Hum Mutat 30:1,2, 2009. © 2009 Wiley-Liss, Inc. [source]

Rapid screening assay of congenital adrenal hyperplasia by measuring 17,-hydroxyprogesterone with high-performance liquid chromatography/electrospray ionization tandem mass spectrometry from dried blood spots

JOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 1 2002
Chien-Chen Lai
Abstract A rapid, simple, and specific method was developed for the diagnosis of congenital adrenal hyperplasia (CAH) from dried blood spots on newborn screening cards based on high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS). The usefulness of 17,-hydroxyprogesterone (17OH-P) determination on dried filter-paper blood samples from patients with CAH caused by 21-hydroxylase deficiency was evaluated. The LC/MS/MS detection of 17OH-P was rapid, <4 min. The intra- and interday accuracy and precision of the method were <7%. Our procedure maintained good linearities (R2 > 0.992) and recovery rate (>83%). We used this new method to directly determine the 17OH-P levels in dried blood specimens from abnormal children of various ages, with a detection limit of 20 ng/ml (,240 pg), to avoid the time-consuming derivatization steps required by the gas-chromatography/mass spectrometry (GC/MS) method. Four dried filter-paper blood samples of CAH patients (three girls and one boy, 1,14 years old) were all quantified in an LC/MS/MS study and revealed high 17OH-P levels (>90 ng/ml). After treatment, all of the elevated 17OH-P levels either decreased or disappeared. Compared with CAH patients, 17OH-P was nearly undetectable (<20 ng/ml) in the normal infants by LC/MS/MS. This LC/MS/MS assay is not only useful for both diagnosis and monitoring of treatment of CAH in all other age groups, it also can be used as a screening test for CAH infants. In this study, we provided the first data on 17OH-P in dried blood specimens affected with CAH using HPLC/ESI-MS/MS. J. Clin. Lab. Anal. 16:20,25, 2002. © 2002 Wiley-Liss, Inc. [source]

Thiopurine S -methyltransferase (TPMT) genetic polymorphisms in Mexican newborns

JOURNAL OF CLINICAL PHARMACY & THERAPEUTICS, Issue 6 2009
A. González-del Angel MD
Abstract Background:, Thiopurine S -methyltransferase (TPMT) is involved in the toxicity and therapeutic efficacy of thiopurine drugs, and its gene exhibits genetic polymorphisms that differ across diverse populations. Four TPMT polymorphisms (TPMT*2, *3A, *3B and *3C) account for 80,95% of alleles that cause reduced enzyme activity. To date, only a single study in the Mexican population involving 108 individuals has been performed, but the regional and ethnic origin of this population was not described. Accordingly, information about the TPMT polymorphism in the Mexican population is limited. Objective:, To determine the TPMT allele and genotype frequencies in a sample of newborns from Mexico City. Methods:, Three hundred and sixty DNA samples from unrelated, anonymous individuals were obtained from dried blood spots collected on filter paper as part of the Newborn Screening National Program. Allele-specific polymerase chain reaction for the TPMT*2 allele and PCR restriction fragment length polymorphism for TPMT*3A, TPMT*3B, TPMT*3C alleles were used to determine the respective allelic and genotypic frequencies. Results and Discussion:, Of 720 TPMT alleles analysed, 49 (6·81%) were deficiency alleles. The most common deficiency allele was TPMT*3A (5·69%), followed by TPMT*3C (0·56%), TPMT*3B (0·28%) and TPMT*2 (0·28%). Fourty-five newborns were heterozygous for one mutant allele (12·5%) and two showed a genotype with two deficiency alleles (0·56%). Despite its unique ethnic composition, our Mexican population exhibited variant allele frequencies that were similar to some Caucasian populations. Conclusion:, Our data suggest that approximately 1 in 180 persons born in Mexico City might have low or undetectable TPMT enzyme activity, a frequency that, overall, is somewhat higher than that reported for Caucasian populations generally (1 in 300). [source]

Glutamic acid decarboxylase and IA-2 autoantibodies in type 1 diabetes: comparing sample substrates for autoantibody determinations

PEDIATRIC DIABETES, Issue 1 2000
C Wasserfall
Large-scale programs designed to assess risk for type 1 diabetes through serologic assessment of autoantibodies to recombinant ,-cell autoantigens are hampered by several limitations, including the methods for sample collection and assay performance, as well as the volume required for autoantibody determinations. The present study was designed to develop a low sample-volume, primary screening method for autoantibody detection of high specificity and sensitivity, and to determine the feasibility of dried blood spots collected on filter paper in serving as vehicles for such determinations. Autoantibodies to glutamic acid decarboxylase (GAD) and ICA512bdc (IA-2), both individually and in combination, were determined in persons with type 1 diabetes, healthy controls, or individuals with other autoimmune disorders. Autoantibody results for serum, plasma, and dried blood spots were compared. GAD, IA-2, and combined GAD/IA-2 autoantibodies were concordant in their measurement from minimal volumes of serum, plasma, and whole blood extracted from dried filter paper. The autoantibody levels from the dried blood spots were, however, lower than corresponding serum samples, and, as currently designed, failed to detect low-titer autoantibodies. Despite this limitation, screening for diabetes risk can be performed using small volumes of whole blood, serum, or plasma collected onto filter paper. These methodological improvements should simplify matters, reduce costs, and increase the efficacy of screening programs for type 1 diabetes. Further development of better substrates/methods for blood-specimen collection seems necessary to exploit the full potential of this and other autoantibody measurement strategies for screening large populations. [source]

Determination of total homocysteine in dried blood spots using high performance liquid chromatography for homocystinuria newborn screening

PEDIATRICS INTERNATIONAL, Issue 1 2004
Andi Dwi Bahagia Febriani
AbstractBackground: The most widely used method for newborn screening for homocystinuria (HCU) is a semiquantitative bacterial inhibition assay for measuring methionine concentration in dried blood spots (DBS). Because this method has resulted in a number of missed cases due to many factors, we developed a high performance liquid chromatography (HPLC) method with fluorescence detection to measure total homocysteine (tHcy) in DBS which might be useful for newborn screening for HCU. Methods: One disk of DBS 3 mm in diameter was sonicated in 10 min. The extract was reduced with dithioerythritol and was derivatized with 4-aminosulfonyl-7fluoro-2,1,3-benzoxadiazole before injection into HPLC. Results: This method showed good linearity (r = 0.996), precision (coefficient of variation range 2.7,5%), and excellent correlation coefficient between DBS and serum tHcy, both in control (r = 0.932) and patient samples (r = 0.952). By this method, the mean tHcy concentration in DBS of preterm newborns, full-term newborns, and adults was 1.4 ± 1.0, 2.5 ± 1.6, and 4.9 ± 1.5 µmol/L, respectively. The mean tHcy DBS concentration in two cases of cystathionine-,-synthase deficiency and one case of 5,10-methylentetrahydrofolate reductase deficiency was 22.7 ± 2.88, 29.3 ± 1.90, and 41.3 µmol/L, respectively. Conclusions: The present method, which is rapid, user friendly and reliable, seems applicable to newborn screening of HCU in place of methionine measurement. [source]

Enzyme immunoassay for total immunoglobulin E in dried blood spots

AMERICAN JOURNAL OF HUMAN BIOLOGY, Issue 3 2007
Susan Tanner
Elevated circulating levels of total immunoglobulin E (IgE) are associated with both allergic disease and repeated macro-parasitic infections. Population-based research on IgE has been limited by the logistical constraints associated with obtaining and processing venipuncture blood samples. In this short report, we present an enzyme immunoassay protocol for quantifying circulating total IgE levels in capillary whole blood, collected from a finger prick and dried on filter paper. The assay demonstrated acceptable levels of accuracy, precision, and reliability. IgE remained stable at room temperature for only 2,4 days and degraded rapidly at higher temperatures suggesting that samples should be refrigerated or frozen within 1,2 days of collection. It is hoped that the relative ease of blood spot collection will expand opportunities for population-based research on IgE.Am. J. Hum. Biol. 19:440,442, 2007. © 2007 Wiley-Liss, Inc. [source]

Direct tandem mass spectrometric analysis of amino acids in dried blood spots without chemical derivatization for neonatal screening

RAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 9 2003
Kornél Nagy
Neonatal screening performed by electrospray tandem mass spectrometry is a powerful technique in clinical diagnostics. In the present paper an alternative to the widely accepted method involving butylation has been developed. In the new method butylation is not required, and multiple reaction monitoring (MRM) was used instead of constant neutral loss scanning. The method was optimized for detection of 23 L-amino acids in their native form. Quantitation was based on isotope-labeled internal standards, calibration curves were linear from 0 to 500,,mol/L, and detection limits were in the range 2,42,,mol/L. The utility of the present technique is illustrated in the case of one neonate suffering from citrullinaemia. Copyright © 2003 John Wiley & Sons, Ltd. [source]

Infants with late breast milk acquisition of HIV-1 generate interferon-gamma responses more rapidly than infants with early peripartum acquisition

CLINICAL & EXPERIMENTAL IMMUNOLOGY, Issue 3 2009
B. Lohman-Payne
Summary Infants infected with HIV-1 after the first month of life have a lower viral set-point and slower disease progression than infants infected before 1 month. We investigated the kinetics of HIV-1-specific CD8+ T lymphocyte secretion of interferon (IFN)-, in infants infected before 1 month of life compared with those infected between months 1 and 12 (late infection). HIV-1 infection was assessed at birth and at months 1, 3, 6, 9 and 12 and timing of infection was determined by HIV-1 gag DNA from dried blood spots and verified by plasma HIV-1 RNA levels. HIV-1 peptide-specific IFN-, responses were measured by enzyme-linked immunospot at months 1, 3, 6, 9 and 12. Timing of development of IFN-, responses was compared using the log,rank test and Kaplan,Meier survival curves. Infants infected late developed HIV-1-specific CD8+ T cell responses 2·8 months sooner than infants infected peripartum: 2·3 versus 5·1 months after HIV-1 infection (n = 52, P = 0·04). Late-infected infants had more focused epitope recognition than early-infected infants (median 1 versus 2 peptides, P = 0·03); however, there were no differences in the strength of IFN-, responses. In infants infected with HIV-1 after the first month of life, emergence of HIV-1-specific CD8+ IFN-, responses is coincident with the decline in viral load, nearly identical to what is observed in adults and more rapid than in early-infected infants. [source]

Evaluation of the Murex HIV Ag/Ab Combination assay when used with dried blood spots

CLINICAL MICROBIOLOGY AND INFECTION, Issue 11 2007
V. Lakshmi
Abstract This study evaluated the ability of the Murex HIV Ag/Ab Combination assay to detect human immunodeficiency virus (HIV) antibodies in 12 617 dried blood spots (DBSs) on filter paper. The assay had an overall sensitivity of 99.6% and a specificity of 99.9%. In view of its ability to detect p24 antigen and both HIV-1 and HIV-2 antibodies in samples collected in the form of DBSs, the Murex Ag/Ab Combination assay is suitable for use as a standard screening assay for seroprevalence studies, as well as for routine diagnostic use in clinical laboratories. [source]