实时定量PCR分析健康人外周血线粒体DNA 4977 bp缺失  

Quantification of 4977 bp deletion in mitochondrial DNA of healthy adults using real-time PCR

作　　者：陈晓培[1] 王平[2] 赵阳辉[1] 韩林[2] 王喜爱[2] 吕玉民[1] 

机构地区：[1]郑州大学公共卫生学院,郑州450001 [2]河南省职业病防治研究所,郑州450052

出　　处：《中国卫生检验杂志》2007年第7期1186-1188,1209,共4页Chinese Journal of Health Laboratory Technology

基　　金：河南省医学科技创新项目(2005077)

摘　　要：目的:用实时定量PCR方法分析健康人外周血线粒体DNA4 977 bp的缺失情况。方法:收集27例年龄在17～44岁的健康人外周血,用荧光实时定量PCR方法对每个样品同时检测线粒体DNA 4 977 bp缺失的量和无4 977 bp缺失的量,进行定量分析。结果:在27例健康人外周血样品中,线粒体DNA4 977 bp缺失的阳性率为70.37%(19/27);19例有线粒体DNA4 977 bp缺失的样品,其扩增产物的CT值在第35个循环到第39个循环之间;无线粒体DNA 4 977 bp缺失的扩增产物的CT值在第15个循环到第21个循环之间;不同年龄组间线粒体DNA 4 977 bp缺失和无线粒体DNA4 977 bp缺失的量均无统计学差异。结论:本文中超过2/3的健康人外周血含有线粒体DNA 4 977 bp的缺失;在有该缺失的样品中大约每106～107拷贝中携带有一个线粒体DNA4 977 bp缺失;随年龄的增长外周血中线粒体DNA 4 977 bp的缺失未显示出累积现象。Objective：To analyze the mitochondrial DNA 4 977 bp deletion in healthy adults by real - time PCR. Methods： The peripheral blood of twenty - seven healthy adults aged from 17 to 44, was collected. The deleted and undeleted mitochondrial DNA in each sample were detected and quantified by real -time PCR. Results：The nineteen samples contained mitochondrial DNA 4 977 bp deletion in all the samples, and the positive rate of the DNA 4 977 bp deletion was 70. 37% （19/27）. The detection threshold（CT） of the amplification products of undeleted mitochondrial DNA varied between the 15th and 21th cycle, and that of the deletion - specific fragments between the 35th and 39th cycle. Amount of the deleted mitochondfial DNA was not statistically significant in different age group, as well as that of undeleted mitochondrial DNA. Conclusion： The peripheral blood of a- bout 2/3 healthy adults contains mitochondrial DNA 4 977 bp deletion in the research. Approximately 1 in 10^6 to 10^7 copies carties the mitochondfial DNA 4 977 bp deletion in the samples containing the deletion. The data does not show any accumulation of the DNA 4 977 bp deletion with increase in age.

关 键 词：线粒体DNA 4977BP缺失 年龄 实时定量PCR 

分 类 号：Q7[生物学—分子生物学]