时相温度梯度凝胶电泳在线粒体基因突变检测中的应用

The application of temporal temperature gradient gel electrophoresis in the detection of mitochondrial gene mutations

目的 评价时相温度梯度凝胶电泳技术 (TTGE)在肿瘤患者线粒体基因同质性和异质性突变检测中的应用。方法 对117例肿瘤组织和配对正常组织的全线粒体基因进行PCR扩增和TTGE突变筛选分析 ,对TTGE显示在肿瘤和配对正常组织中具有不同类型电泳带型变化的片段进行测序。结果 TTGE在总共 36 5 4片段中发现 16 0个体细胞性线粒体基因突变片段 ,经与直接测序结果比较 ,TTGE检测体细胞性突变的敏感性和特异性分别达 96 2 7%和 94 0 5 %。结论 TTGE是筛选线粒体基因体细胞性同质性突变和各种比例异质性突变的一种敏感方法。

Objective To evaluate the potential utility of temporal temperature gradient gel electrophoresis (TTGE) in detecting homoplasmic and heteroplasmic mtDNA mutations in patients with cancers. Methods The entire mtDNA of tumor and corresponding normal tissues from total 117 patients were amplified by PCR, and the somatic mutations were screened by TTGE. DNA fragments showing different band patterns in normal and tumor tissues in TTGE were sequened to identify the exact mutations. Results 160 fragments harbouring mtDNA mutations were found by TTGE in total 3 654 fragments. Compared with the direct squencing results, the sensitivity and specificity of TTGE in mutation detection were 96.27% and 94.05% respectively. Conclusion TTGE is an effective method for screening acquired homoplasmic mitochondrial gene mutation and its different percentage.