Department of Psychiatry and Behavioral Sciences, School of Medicine, Tulane University, New Orleans, Louisiana, United States of America.
Tulane Brain Institute, Tulane University, New Orleans, Louisiana, United States of America.
PLoS One. 2021 Jan 20;16(1):e0245582. doi: 10.1371/journal.pone.0245582. eCollection 2021.
Use of telomere length (TL) as a biomarker for various environmental exposures and diseases has increased in recent years. Various methods have been developed to measure telomere length. Polymerase chain reaction (PCR)-based methods remain wide-spread for population-based studies due to the high-throughput capability. While several studies have evaluated the repeatability and reproducibility of different TL measurement methods, the results have been variable. We conducted a literature review of TL measurement cross-method comparison studies that included a PCR-based method published between January 1, 2002 and May 25, 2020. A total of 25 articles were found that matched the inclusion criteria. Papers were reviewed for quality of methodologic reporting of sample and DNA quality, PCR assay characteristics, sample blinding, and analytic approaches to determine final TL. Overall, methodologic reporting was low as assessed by two different reporting guidelines for qPCR-based TL measurement. There was a wide range in the reported correlation between methods (as assessed by Pearson's r) and few studies utilized the recommended intra-class correlation coefficient (ICC) for assessment of assay repeatability and methodologic comparisons. The sample size for nearly all studies was less than 100, raising concerns about statistical power. Overall, this review found that the current literature on the relation between TL measurement methods is lacking in validity and scientific rigor. In light of these findings, we present reporting guidelines for PCR-based TL measurement methods and results of analyses of the effect of assay repeatability (ICC) on statistical power of cross-sectional and longitudinal studies. Additional cross-laboratory studies with rigorous methodologic and statistical reporting, adequate sample size, and blinding are essential to accurately determine assay repeatability and replicability as well as the relation between TL measurement methods.
近年来，端粒长度（TL）作为各种环境暴露和疾病的生物标志物的使用有所增加。已经开发了各种方法来测量端粒长度。由于高通量能力，基于聚合酶链反应（PCR）的方法在基于人群的研究中仍然广泛使用。虽然已经有几项研究评估了不同 TL 测量方法的可重复性和再现性，但结果各不相同。我们对包括 2002 年 1 月 1 日至 2020 年 5 月 25 日发表的基于 PCR 的方法在内的 TL 测量跨方法比较研究进行了文献回顾。共发现符合纳入标准的 25 篇文章。对方法报告的样本和 DNA 质量、PCR 分析特征、样本盲法以及确定最终 TL 的分析方法进行了质量评估。总体而言，按照两种不同的基于 qPCR 的 TL 测量报告指南，方法报告的质量较低。方法之间的相关性（由 Pearson's r 评估）差异很大，很少有研究使用推荐的组内相关系数（ICC）来评估测定的重复性和方法比较。几乎所有研究的样本量都小于 100，这引起了对统计功效的关注。总体而言，本综述发现，目前关于 TL 测量方法之间关系的文献在有效性和科学严谨性方面存在不足。鉴于这些发现，我们提出了基于 PCR 的 TL 测量方法的报告指南，并分析了测定重复性（ICC）对横断面和纵向研究统计功效的影响。额外的实验室间研究需要严格的方法学和统计学报告、足够的样本量和盲法，以准确确定测定的重复性和可重复性以及 TL 测量方法之间的关系。
