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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 application of time-gradient temperature gradient gel electrophoresis (TTGE) in the detection of mitochondrial DNA homogeneity and heterogeneity in cancer patients. Methods The full-length mitochondrial DNA of 117 tumor tissues and paired normal tissues was amplified by PCR and the TTGE mutation screening was performed. Sequences with different types of electrophoresis bands in the tumor and paired normal tissues were sequenced. Results TTGE found 160 somatic mitochondrial DNA mutations in a total of 36 5 4 fragments. Compared with the direct sequencing results, the sensitivity and specificity of TTGE in detecting somatic mutations were 96 27% and 94 05% . Conclusion TTGE is a sensitive method for screening somatic homozygous mitochondrial DNA mutations and heterogeneous mutations of various proportions.