荧光定量PCR测定端粒长度

目的应用实时荧光定量聚合酶链式反应（Q—PCR）方法测定端粒长度。方法选取9种人类细胞株，提取基因组DNA，采用Q—PCR方法测定相对T／S比率，DNA印迹法测定末端限制性片段（TRF）长度，进行二者之间的相关性分析。结果定量PCR测定端粒长度相对T／S比率为0．68±0．57，DNA印迹法测量平均TRF值为8．57±2．34，两种方法测定结果的相关性分析R2=0．7807（P〈0．01）。结论采用荧光定量PCR方法测量端粒长度具有重复性好、省时、简便、可靠的特点，可高通量处理大量样品。

Telomere Measurement by Quantitative PCR

Objective To determine relative telomere length by quantitative PCR. Methods Genomic DNA was extracted from 9 types of human ceils. Relative T/S ratios measured by quantita- tive PCR were compared with relative mean terminal restriction fragment （TRF） lengths in the same samples measured by traditional Southern blot methods. Results Relative R/S ratio measured by Q-PCR was 0.68 ＋ O. 57, and relative mean TRF length was 8.57 ±2.34. By linear regression analysis, the correlation coefficient, R2, for the relationship of T/S ratio to TRF length, .was 0. 7807 （P 〈 0. 001 ） . Conclusion We have demonstrated the measurement of relative average te- lomere lengths by quantitative PCR, using a carefully designed pair of oligonucleotide primers. The assay is simple, rapid and reproducible, thus reliable for a high throughput of samples. It is recom- mendable to be used in study of telomere biology and genetic epidemiology of cancer and aging related diseases.