Comparison of telomere length measurement methods

Abstract

The strengths and limitations of the major methods developed to measure telomere lengths (TLs) in cells and tissues are presented in this review. These include Q-PCR (uantitative olymerasehain eaction), TRF (erminal estrictionragment) analysis, a variety of Q-FISH (uantitative luorescencen ituybridization) methods, STELA (ingle lomereength nalysis) and TeSLA (lomere hortestength ssay). For each method, we will cover information about validation studies, including reproducibility in independent laboratories, accuracy, reliability and sensitivity for measuring not only the average but also the shortest telomeres. There is substantial evidence that it is the shortest telomeres that trigger DNA damage responses leading to replicative senescence in mammals. However, the most commonly used TL measurement methods generally provide information on average or relative TL, but it is the shortest telomeres that leads to telomere dysfunction (identified by TIF,elomere dysfunction nducedoci) and limit cell proliferation in the absence of a telomere maintenance mechanism, such as telomerase. As the length of the shortest telomeres is a key biomarker determining cell fate and the onset of senescence, a new technique (TeSLA) that provides quantitative information about all the shortest telomeres will be highlighted.This article is part of the theme issue 'Understanding diversity in telomere dynamics'.

Ethnic disparities in relationships of obesity indices with telomere length in Asians with type 2 diabetes

“…Second, the sample sizes for Malays and Indians are small, which may increase the possibility that the results in these ethnic groups were due to chance. Third, quantitative PCR is a widely used and cost‐efficient high‐throughput technique, but mean bulk TL does not capture subpopulations of cells with abnormally short telomeres . There may also be changes in the distribution of cell types at baseline and follow‐up .…”

“…Third, quantitative PCR is a widely used and cost-efficient high-throughput technique, but mean bulk TL does not capture subpopulations of cells with abnormally short telomeres. 42 There may also be changes in the distribution of cell types at baseline and follow-up. 43 Furthermore, telomere length was measured in blood cells and not in adipose tissues.…”

Telomere length regulators are activated in young men after pediatric kidney transplantation compared to healthy controls and survivors of childhood cancer—A cross‐sectional study

“…A methodological caveat of the study was that telomere length was measured using the q‐PCR method. Q‐PCR has been shown to be a relatively inaccurate method in the case of measuring both extremely short and long telomere length …”

“…Q-PCR has been shown to be a relatively inaccurate method in the case of measuring both extremely short and long telomere length. 35 The strength of our study is that all subjects were carefully examined and documented. We also believe that although our study population represents only a relatively small proportion of all transplanted boys in Finland, it does represent the national cohort based on hospital records and data from the Finnish Registry for Kidney Diseases.…”

Understanding diversity in telomere dynamics

“…Of course our understanding of telomere diversity is very much dependent on obtaining accurate measures of telomere length and loss, and this is fraught with difficulties particularly when using non-model species [17]. The paper by Lai et al [18] discusses these difficulties, and the problem introduced when examining links to disease and/or longevity when estimating average telomere length rather than the shortest telomeres.…”