荧光定量PCR测定端粒长度  

Telomere Measurement by Quantitative PCR

作　　者：王敏敏[1] 杨浩[1] 刘广义[1] 陈大进[1] 陈文清[1] 蔡洁茹[1] 周芹[1] 谢易[1] 陈芷珉[1] 陈莹[1] 王慧萍[1] 冯时[1] 王宇成[1] 茅幼英[1] 姜虹[1] 陈江华[1] 

机构地区：[1]浙江大学医学院附属第一医院肾脏病中心,杭州市310003

出　　处：《医学分子生物学杂志》2009年第6期494-497,共4页Journal of Medical Molecular Biology

基　　金：资助项目：国家自然科学基金（No.30671990,30900689）

摘　　要：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。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.

关 键 词：荧光定量聚合酶链式反应 端粒长度测定 T/S比率 

分 类 号：R73-34[医药卫生—肿瘤]