Measuring the TREC ratio in dried blood spot samples: Intra- and inter-filter paper cards reproducibility

P. O. Lang, S. Govind, M. Dramé, R. Aspinall

Research output: Contribution to journalArticle

6 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1-8
Number of pages8
JournalJournal of Immunological Methods
Volume389
Issue number1-2
Early online date9 Jan 2013
DOIs
Publication statusPublished - 29 Mar 2013
Externally publishedYes

Keywords

  • Dried blood spots
  • Quantitative PCR
  • Real time PCR
  • Reproducibility
  • Thymic TREC
  • TREC ratio

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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