用错配PCR技术直接检测母系遗传性非综合征型聋相关线粒体DNA C1494T突变  

Detecting C1494 Tmutation associated with maternally transmitted aminoglycoside-induced and nonsyndromic deafness using mispairing PCR

作　　者：刘琪[1,2,3] 赵辉[3] 

机构地区：[1]绍兴市人民医院 [2]浙江大学绍兴医院检验科,浙江绍兴312000 [3]解放军总医院耳鼻咽喉头颈外科,北京100853

出　　处：《中国耳鼻咽喉头颈外科》2012年第3期134-136,共3页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：国家自然科学基金面上项目(30600701);全军"十一五"课题青年学者项目(06Q072);教育部留学回国人员科研启动基金资助(赵辉)项目;高等学校全国优秀博士学位论文作者专项基金(2007B6)联合资助

摘　　要：目的在一个聚合酶链式反应(polymerasechain reaction,PCR)管中运用错配PCR技术直接检测母系遗传性非综合征型聋相关的线粒体DNA C1494T突变。方法设计一对引物对母系遗传性非综合征型聋相关的线粒体DNA C1494T突变的单碱基错配引物,即在引物的3'端附近引入一个与模板错配的碱基。用此引物进行多聚酶链式反应,再通过琼脂糖凝胶电泳观察结果。结果通过电泳结果可以直观判断患者是否携带线粒体DNA C1494T突变,结果与测序结果一致。结论错配PCR技术可以准确、方便、快速检测线粒体DNA C1494T突变,适于在基层开展和应用。OBJECTIVE To detect C1494T mutation associated with maternally transmitted aminoglycosideinduced and nonsyndromic deafness in one pipe with mispairing PCR.METHODS A pair of mispairing primer according to mitochondrial DNA C1494T mutation was designed.PCR was carried out with designed primer,and the results were observed through agarose gel electrophoresis.RESULTS We could judge mitochondrial DNA C1494T mutation easily,and the results were identical with sequencing.CONCLUSION Mispairing PCR method can accurately,convenient,quickly detect mitochondrial DNA C1494T mutation and is suitable for application in the basic unit.

关 键 词：听力障碍 基因 DNA 线粒体 突变 错配PCR C1494T 

分 类 号：R764[医药卫生—耳鼻咽喉科]