Blood and dried blood spot telomere length measurement by qPCR: assay considerations.
Blood and dried blood spot telomere length measurement by qPCR: assay considerations.
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DOI：
10.1371/journal.pone.0057787
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Côté HC
中科院分区：
文献类型：
--
作者：
Zanet DL;Saberi S;Oliveira L;Sattha B;Gadawski I;Côté HC
端粒长度的测量对于研究端粒维持及其在疾病和衰老的分子病理生理中的作用至关重要。有几种方法用于测量端粒长度，其选择通常取决于要分析的样品的类型和大小，以及成本和吞吐量的考虑。本研究的目的是研究可能影响全血中基于qpcr的相对端粒长度测量可靠性的因素。每天的个体内变异、血液抗凝剂类型、样本储存条件、处理和抽血地点进行了调查。还研究了两种基于qpcr的测量端粒长度的方法（单端粒和多端粒），并显示它们之间存在很强的相关性。血液的冷冻和解冻以及血液在4°C下储存4天对端粒长度值没有影响。干血斑端粒长度显著高于全血和外周血单核细胞，且与全血和外周血单核细胞端粒长度高度相关。我们发现，与从手指采集的抗凝全血和从手指和手臂静脉穿刺采集的未染色全血制备的干血点相比，直接从指尖穿刺采集的干血点的端粒长度测量值明显更高。这表明，从纸上吸取的细胞DNA与从静脉穿刺全血中采集的DNA并不等同，在比较血液样本类型时应谨慎。
Measurement of telomere length is crucial for the study of telomere maintenance and its role in molecular pathophysiology of diseases and in aging. Several methods are used to measure telomere length, the choice of which usually depends on the type and size of sample to be assayed, as well as cost and throughput considerations. The goal of this study was to investigate the factors that may influence the reliability of qPCR-based relative telomere length measurements in whole blood. Day to day intra-individual variability, types of blood anticoagulant, sample storage conditions, processing and site of blood draw were investigated. Two qPCR-based methods to measure telomere length (monoplex vs. multiplex) were also investigated and showed a strong correlation between them. Freezing and thawing of the blood and storage of the blood at 4°C for up to 4 days did not affect telomere length values. Telomere lengths in dried blood spots were significantly higher than both whole blood and peripheral mononuclear blood cells, and were highly correlated with both. We found that telomere length measurements were significantly higher in dried blood spots collected directly from fingertip prick compared to dried blood spots prepared with anticoagulated whole blood collected from the finger, and non-blotted whole blood taken from both finger and arm venipuncture. This suggests that DNA from cells blotted on paper is not equivalent to that collected from venipuncture whole blood, and caution should be taken when comparing between blood sample types.
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DOI：
10.1016/j.mrfmmm.2011.04.003
发表时间：
2012-02-01
影响因子：
2.3
作者：
Aubert, Geraldine;Hills, Mark;Lansdorp, Peter M.
通讯作者：
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DOI：
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发表时间：
2003-02-01
期刊：
CLINICAL AND LABORATORY HAEMATOLOGY
影响因子：
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作者：
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通讯作者：
Gürakan, B
影响因子：
3.8
作者：
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通讯作者：
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影响因子：
3.7
作者：
Halfon P;Ouzan D;Khiri H;Pénaranda G;Castellani P;Oulès V;Kahloun A;Amrani N;Fanteria L;Martineau A;Naldi L;Bourlière M
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Bourlière M