Assessment of intra- and inter-assay variation in dried blood spot telomere length measurements by qPCR

抄録

Telomere length has recently gained popularity as a biomarker of aging-related diseases. Dried blood spot (DBS) samples are commonly used to measure telomere length for human biology research, but studies assessing intra- and inter-assay variations are scarce. The objective of the present study was to analyze and elucidate the extent of intra- and inter-assay variation of DBS telomere length measurement. The DBS telomere length of a male subject was determined by quantitative polymerase chain reaction (qPCR) using four different protocols with two different container types (96-well plates and 8-well tubes) and two different primer sets (tel1b–tel2b and tel1–tel2). The measurement of telomere length with 8-well tubes and the tel1–tel2 primer set demonstrated the lowest intra-assay coefficient of variation (CV) (6.0%) and gave an inter-assay CV of 5.0%. The protocol for 96-well plates typically resulted in high intra-assay CVs (>27%). Assessment of intra-assay variation is essential when DBS telomere length is measured by qPCR.