The Center of Human development and Aging, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, NJ 07103, USA.
Nucleic Acids Res. 2011 Nov 1;39(20):e134. doi: 10.1093/nar/gkr634. Epub 2011 Aug 8.
Telomere length/DNA content has been measured in epidemiological/clinical settings with the goal of testing a host of hypotheses related to the biology of human aging, but often the conclusions of these studies have been inconsistent. These inconsistencies may stem from various reasons, including the use of different telomere length measurement techniques. Here, we report the first impartial evaluation of measurements of leukocyte telomere length by Southern blot of the terminal restriction fragments and quantitative PCR (qPCR) of telomere DNA content, expressed as the ratio of telomeric product (T)/single copy gene (S) product. Blind measurements on the same samples from 50 donors were performed in two independent laboratories on two different occasions. Both the qPCR and Southern blots displayed highly reproducible results as shown by r values > 0.9 for the correlations between results obtained by either method on two occasions. The inter-assay CV measurement for the qPCR was 6.45%, while that of the Southern blots was 1.74%. The relation between the results generated by Southern blots versus those generated by qPCR deviated from linearity. We discuss the ramifications of these findings with regard to measurements of telomere length/DNA content in epidemiological/clinical circumstances.
端粒长度/DNA 含量已在流行病学/临床环境中进行了测量，目的是测试与人类衰老生物学相关的一系列假设，但这些研究的结论往往不一致。这些不一致可能源于各种原因，包括使用不同的端粒长度测量技术。在这里，我们报告了首次对端粒末端限制片段 Southern 印迹和端粒 DNA 含量的定量 PCR（qPCR）测量白细胞端粒长度的公正评估，端粒 DNA 含量表示为端粒产物（T）/单拷贝基因（S）产物的比值。在两个不同的场合，在两个独立的实验室对来自 50 个供体的相同样本进行了盲测。qPCR 和 Southern blot 的结果都显示出高度可重复性，两种方法在两个场合获得的结果之间的 r 值均大于 0.9。qPCR 的批内变异系数测量值为 6.45%，而 Southern blot 的为 1.74%。Southern blot 产生的结果与 qPCR 产生的结果之间的关系偏离线性。我们讨论了这些发现对流行病学/临床情况下端粒长度/DNA 含量测量的影响。
