单色多重荧光实时定量PCR测定端粒长度

Telomere length measurement by monochrome multiplex quantitative PCR method

目的在罗氏Light Cycler 480上建立单色多重荧光实时定量聚合酶链反应(MMQPCR)方法测定端粒长度。方法在罗氏480定量PCR仪上使用MMQPCR方法测定39例人外周血白细胞端粒长度(相对T/S比值),并与DNA印迹法(Southern blot)测得的平均末端限制性片段(TRF)长度作比较。结果 MMQPCR方法测定端粒长度相对T/S比值为1.13±0.21,Southern blot方法测量平均TRF长度为(7.46±1.21)kb,两种方法测定结果的相关性分析R^2=0.6706(P<0.001)。结论本研究建立的MMQPCR方法测定端粒长度重复性好、省时、简便、可靠,可高通量处理大量样品。

Objective To establish a monochrome multiplex real-time quantitative polymerase chain reaction （MMQPCR）, for telomere length measurement on the Roche LightCycler 480 （LC480） real-time PCR platform. Methods Telomere lengths （T/S ratio） were measured from 39 DNA samples extracted from human white blood cells using the MMQPCR method on the LC480 platform, and were compared with terminal restriction fragment （TRF） lengths measured by Southern blot. Results Relative T/S ratio measured by MMQPCR was 1.13±0.21, and mean TRF length was （7.46±1.21） kb. The correlation coefficient （R^2） for the relationship of the two different methods was 0.6706 （P 〈 0.001）. Conclusion The MM- PQPCR method is reproducible, rapid, and simple, thus reliable for a high throughput of samples.