Measuring the TREC ratio in dried blood spot samples: intra- and inter-filter paper cards reproducibility.
Details
Serval ID
serval:BIB_F14CB9CCEA93
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Measuring the TREC ratio in dried blood spot samples: intra- and inter-filter paper cards reproducibility.
Journal
Journal of Immunological Methods
ISSN
1872-7905 (Electronic)
ISSN-L
0022-1759
Publication state
Published
Issued date
2013
Peer-reviewed
Oui
Volume
389
Number
1-2
Pages
1-8
Language
english
Notes
Publication types: Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Abstract
The level of T-cell receptor excision circles (TREC), which decline with advancing age in normal individuals, has recently gained interest as a reference marker for studies on premature or early immunosenescence under particular health conditions. In order to facilitate translational studies at population and clinical levels, essential for the understanding of how changes in TREC levels are associated with responsiveness of the immune system, we have developed and optimized a real-time polymerase chain reaction (qPCR) assay which quantifies the TREC ratio from dried blood spot (DBS) samples. The present study considers a fully automated procedure to purify DNA and amplify sequences of interests by means of qPCR from DBS samples collected in healthy adults. Both TREC:PBMC (peripheral blood mononuclear cell) and TREC:T-cell ratios were compared for intra- and inter individual reproducibility. Furthermore, the impact of the length of storage on the quality of the DNA generated was also analyzed. In conclusion we describe a fully automated procedure for extracting DNA and qPCR set up, which offers a high-precision, robust qPCR assay for the quantification of both TREC:T-cell ratio and TREC:PBMC from DBS samples.
Keywords
Adult, DNA, Circular/blood, DNA, Circular/chemistry, Dried Blood Spot Testing/instrumentation, Dried Blood Spot Testing/methods, Female, Humans, Immunity, Innate/immunology, Leukocytes, Mononuclear/chemistry, Leukocytes, Mononuclear/immunology, Male, Real-Time Polymerase Chain Reaction/methods, Real-Time Polymerase Chain Reaction/standards, Receptors, Antigen, T-Cell/blood, Receptors, Antigen, T-Cell/genetics, Reproducibility of Results, T-Lymphocytes/chemistry, T-Lymphocytes/immunology
Pubmed
Web of science
Create date
15/04/2015 8:14
Last modification date
20/08/2019 16:18