Telomere length has been used as a proxy of fitness, aging and lifespan in vertebrates. In the last decade, dozens of articles reporting on telomere dynamics in the fields of ecology and evolution have been published for a wide range of taxa. With this growing interest, it is necessary to ensure the accuracy and reproducibility of telomere length measurement techniques. Real-time quantitative PCR (qPCR) is routinely applied to measure relative telomere length. However, this technique is highly sensitive to several methodological variables and the optimization of qPCR telomere assays remains highly variable between studies. Therefore, standardized guidelines are required to enable the optimization of robust protocols, and to help in judging the validity of the presented results. This review provides an overview of preanalytical and analytical factors that can lead to qPCR inconsistencies and biases, including: (a) sample type, collection and storage; (b) DNA extraction, storage and quality; (c) qPCR primers, laboratory reagents, and assay conditions; and (d) data analysis. We propose a minimum level of information for publication of qPCR telomere assays in evolutionary ecology considering the methodological pitfalls and sources of error. This review highlights the complexity of the optimization and validation of qPCR for telomere measurement per se, demonstrating the importance of transparency and clarity of reporting methodological details required for reliable, reproducible and comparable qPCR telomere assays. We encourage efforts to implement standardized protocols that ensure the rigour and quality of telomere dynamics studies.

中文翻译：

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端粒定量PCR公开的标准指南导致了进化生态学。

端粒的长度已被用作脊椎动物的适应性，衰老和寿命的代表。在过去的十年中，已经发表了许多有关生态和进化领域端粒动力学的文章，涉及广泛的分类单元。随着这种日益增长的兴趣，有必要确保端粒长度测量技术的准确性和可重复性。实时定量PCR（qPCR）通常用于测量相对端粒长度。但是，该技术对几种方法学变量高度敏感，qPCR端粒测定的优化在研究之间仍然存在很大差异。因此，需要标准化的准则来实现健壮协议的优化，并有助于判断所提供结果的有效性。这篇综述概述了可能导致qPCR不一致和偏差的分析前和分析因素，包括：（a）样品类型，收集和储存；（b）DNA的提取，储存和质量；（c）qPCR引物，实验室试剂和测定条件；（d）数据分析。考虑到方法上的陷阱和错误来源，我们提出了在进化生态学中发布qPCR端粒测定的最低信息水平。这篇综述强调了端粒测量本身优化和验证qPCR的复杂性，证明了透明，透明的报告可靠性，可重复性和可比性qPCR端粒测定所需的方法学细节的重要性。