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Telomere length measurement by qPCR in birds is affected by storage method of blood samples

机译:通过qpCR测定鸟类中的端粒长度受血液样品的储存方法的影响

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摘要

Given the potential role of telomeres as biomarkers of individual health and ageing, there is an increasing interest in studying telomere dynamics in a wider range of taxa in the fields of ecology and evolutionary biology. Measuring telomere length across the lifespan in wild animal systems is essential for testing these hypotheses, and may be aided by archived blood samples collected as part of longitudinal field studies. However, sample collection, storage, and DNA extraction methods may influence telomere length measurement, and it may, therefore, be difficult to balance consistency in sampling protocol with making the most of available samples. We used two complementary approaches to examine the impacts of sample storage method on measurements of relative telomere length (RTL) by qPCR, particularly focusing on FTA (Flinders Technology Associates) cards as a long-term storage solution. We used blood samples from wandering albatrosses collected over 14 years and stored in three different ways (n = 179), and also blood samples from captive zebra finches (n = 30) that were each stored using three different methods. Sample storage method influenced RTL in both studies, and samples on FTA cards had significantly shorter RTL measurements. There was no significant correlation between RTL measured in zebra finch blood on FTA cards and the same samples stored either as frozen whole blood or as extracted DNA. These results highlight the importance of consistency of sampling protocol, particularly in the context of long-term field studies, and suggest that FTA cards should not be used as a long-term storage solution to measure RTL without validation.
机译:考虑到端粒作为个体健康和老龄化的生物标记物的潜在作用,人们越来越关注在生态学和进化生物学领域研究广泛分类单元中端粒动力学的兴趣。测量野生动物系统中整个寿命的端粒长度对于检验这些假设至关重要,并且可以通过纵向田野研究的一部分收集的已归档血液样本进行辅助。但是,样品的收集,存储和DNA提取方法可能会影响端粒长度的测量,因此,可能难以在采样方案的一致性和充分利用可用样品之间取得平衡。我们使用两种互补的方法来检查样本存储方法对通过qPCR测量相对端粒长度(RTL)的影响,尤其着眼于作为长期存储解决方案的FTA(Flinders Technology Associates)卡。我们使用了在14年中收集的流浪信天翁的血液样本,并以三种不同的方式进行存储(n = 179),还使用了圈养斑马雀(n = 30)的血液样本,分别使用三种不同的方法进行了存储。两项研究中,样品存储方法均会影响RTL,FTA卡上的样品的RTL测量值明显较短。在FTA卡上的斑马雀科血液中测得的RTL与以冷冻全血或提取的DNA储存的相同样品之间没有显着相关性。这些结果凸显了采样协议一致性的重要性,尤其是在长期的现场研究中,并且建议FTA卡不应用作未经验证的用于测量RTL的长期存储解决方案。

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