巢式PCR分析电离辐射诱导人外周血线粒体DNA4977bp缺失  

Detection the human mitochondrial DNA 4977 bp deletion induced by ^(60)Co γ-rays in vitro by nest-PCR

作　　者：封江彬[1] 陆雪[1] 陈德清[1] 陈晓穗[2] 刘青杰[1] 

机构地区：[1]中国疾病预防控制中心辐射防护与核安全医学所,北京100088 [2]海军总医院

出　　处：《中华放射医学与防护杂志》2004年第6期533-536,共4页Chinese Journal of Radiological Medicine and Protection

基　　金：人事部留学回国人员择优资助项目 (2 0 0 2年 优秀类 )

摘　　要：目的　探索对不同剂量电离辐射诱导的人外周血线粒体DNA 4 977bp缺失进行分析的方法。方法　采用 3对引物对γ射线诱导的人外周血线粒体DNA缺失进行PCR扩增 ,建立对人线粒体DNA 4 977bp缺失进行分析的巢式PCR方法 ,扩增线粒体DNA缺失区片段 ,纯化PCR产物进行测序。采集 5例正常人外周血进行 0～ 5Gy6 0 Coγ射线的外照射 ,利用巢式PCR技术 ,分析6 0 Coγ射线照射后外周血有核细胞中线粒体DNA 4 977bp缺失的发生情况。结果　用建立的巢式PCR方法可在受到照射的外周血样品中检测到mtDNA 4 977bp缺失。所有外周血样本在照射前均无线粒体DNA 4 977bp缺失 ,1～ 5Gy6 0 Coγ线照射后均诱导出此缺失。结论　本研究所建立的巢式PCR方法可用来分析电离辐射诱导的人外周血淋巴细胞线粒体DNA 4 977bp缺失。Objective To establish a method for detecting the mitochondrial DNA 4977 bp deletion (mtDNA 4977) induced by different doses of ionizing radiation. Methods A nest-PCR method was established with 3 primer pairs for detecting the human peripheral mtDNA 4977.The final PCR products were sequenced after purified and the sequence was BLASTed with the standard genome information of human mitochondrion. The mtDNA 4977 level induced by 05 Gy 60Co γ-rays of 5 healthy individuals was analyzed with the established nest-PCR. Results The mtDNA 4977 could be detected by the established nest-PCR method. The mtDNA 4977 was observed on samples after exposed to 15 Gy 60Co γ-rays,but it was not observed before (0 Gy) exposure. Conclusion The nest-PCR method established in this study could be used to detect the mtDNA 4977 induced by ionizing radiation.;

关 键 词：人外周血 巢式PCR 线粒体DNA 照射 缺失 ^60Coγ射线 电离辐射诱导 PCR产物 测序 纯化 

分 类 号：R818[医药卫生—放射医学]