【摘要】 目的:检测人类线粒体DNA异质性从而建立有效的筛查低水平异质性DNA的方法。方法:选择包含Ⅱ型糖尿病高突变位点的人mtDNA 3083～3339 nt片段进行PCR扩增,以不同比例混合相差一个碱基的两种DNA片段的PCR扩增产物,建立不同比例异质性DNA模型。确定最佳检测条件后分别用变性高效液相色谱技术(DHPLC)和DNA测序技术进行分析和比较。结果:应用DHPLC检测mtDNA 3083～3339 nt(257 bp),最低可检测出该区域含量为5%的异质性样本,而DNA测序技术只能检测出异质性含量大于20%的样本。结论:成功建立分析mtDNA3083～3339 nt片段异质性突变的方法,该方法可用于检测低水平的mtDNA异质性。更多还原

【Abstract】 Objective:To examine the heteroplasmy of human mtDNA, thereby to establish an effective technique to screen low-level heteroplasmic DNAs. Methods: The 257bp PCR products of two kinds of mtDNAs differed from only one alkali base were mixed in different ratios as models. Heteroplasmic mtDNAs were examined with DHPLC and the direct sequencing. The optimal conditions were determined. Results: The least distinguishable ratio of the heteroplasmic DNAs (257 bp fragments) was 5% by DHPLC, but was 20% by the direct sequencing. Conclusion: The established method of PCR-DHPLC can be used for screening the heteroplasmy of mtDNA.更多还原