重复性
复制
端粒
再现性
人口
早晨
生物标志物
样品(材料)
傍晚
医学
生物
统计
数学
内科学
色谱法
化学
遗传学
DNA
环境卫生
物理
天文
作者
Marsha Blauwkamp,Clare L. Fasching,Jingyu Lin,Karl J. Guegler,Evangelos Hytopoulos,Drew Watson,Calvin B. Harley
出处
期刊:The journal of applied laboratory medicine
[Oxford University Press]
日期:2017-07-01
卷期号:2 (1): 4-16
被引量:1
标识
DOI:10.1373/jalm.2016.022137
摘要
Abstract Background Average telomere length in whole blood has become a biomarker of aging, disease, and mortality risk across a broad range of clinical conditions. The most common method of telomere length measurement for large patient sample sets is based on quantitative PCR (qPCR). For laboratory-developed tests to be performed on clinical samples, they must undergo a rigorous analytical validation, currently regulated under CLIA. Methods Whole blood samples from 40 donors were used in the analytical validation of methods for relative average telomere length (rATL) measurement. Three technical replicate DNA samples were extracted from each whole blood sample and placed in three independent wells on a sample plate. Each of these sample plates was assayed 12 times during the validation process. The study was conducted over a 20-day period, once in the morning and once in the evening, using 3 different operators. Results Our process of rATL measurement beginning with DNA extraction followed by qPCR-based assay resulted in repeatability and reproducibility CV of <5% and amplification efficiencies near 100%. The validated assay was used to establish a reference interval derived from 2 cohorts of individuals: (a) San Francisco Bay area (n = 504) and (b) a US cross-sectional, demographic population (n = 357). Conclusions We present advances in the establishment of a highly reproducible analytically validated process for determining rATLs in a CLIA laboratory environment.
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