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Article Abstract

Background: Telomere length (TL) is a valuable marker of aging and stress that reflects both genetic and environmental influences. Quantitative PCR (qPCR) TL measurement is a powerful and cost-effective assay, especially in population studies with limited quantities of source material. Nevertheless, collecting and transporting high-quality blood samples can be logistically challenging, and research suggests that several preanalytical and analytical factors can influence the reliability and precision of the qPCR assay. Here we describe a procedure for collecting blood remotely in a large-scale study. We then assess the influence of various features of the samples, as well as their collection, transportation, and storage on DNA quality and TL assay outcomes.

Method: Participants used at-home collection kits to collect a few drops of whole blood in BD Microtainers during a baseline (n = 265) and 1-year follow-up (n = 178) assessment. DNA was extracted using a magnetic-bead method, and DNA yield, purity, and integrity were assessed. TL was measured using qPCR. To assess inter-assay variation, the coefficient of variation (CV) was calculated across repeated TL measurements (three runs) for each sample. When there was adequate material for duplicate extractions of DNA from the same blood samples, we calculated the intra-class correlation (ICC) of the resultant TL values to assess assay precision.

Results: Our analyses revealed that as little as 50 μL of blood yielded sufficient DNA for highly precise TL measurement (ICC = 0.962, n = 365). Transportation time and an additional year of storage time at -80°C did not meaningfully affect DNA quality or assay outcomes. However, blood clotting was associated with longer telomere estimates, whereas greater temperature exposure was related to shorter telomere estimates.

Conclusions: We established that whole blood collected remotely in BD Microtainers can provide a valid sample source for qPCR TL measurement. We also outline important logistical considerations related to sample collection and handling and provide recommendations for researchers who want to use this method.

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Source
http://dx.doi.org/10.1002/ajhb.70128DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC12413658PMC

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