【摘要】 [目的]检测人类mtDNA HVI区的异质性,建立有效的筛查低水平异质性DNA的技术。[方法]扩增人mtDNA 的 HVI区内269 bp的片段,以不同比例混合相差一个碱基的2种DNA片段的PCR扩增产物,配制异质性DNA模型,用变性高效液相色谱技术(DHPLC)检测,确定最佳检测条件。[结果]应用DHPLC检测HVI区的片段(269 bp),最低可检测出含量为5％的异质性样本。在80例无关个体的mtDNA样本中,共检测到15例具有异质性。[结论]所建立的PCR-DHPLC方法可用于检测mtDNA的异质性。更多还原

【Abstract】 [Objective] To examine the heteroplasmy of hypervariable region I of human mtDNA, and to establish an effective technique to screen low-level heteroplasmic DNAs. [Method] The 269 bp PCR products of the hypervariable region I of 2 kinds of mtDNAs differed from only one alkali base were mixed in different ratios as models. Heteroplasmic mtDNAs were examined by DHPLC. The optimal conditions for testing were determined. The PCR products of the Hypervariable Region I of mtDNAs from 80 individuals were examined. [Result] The least distinguishable ratio of the heteroplasmic DNAs (269 bp fragments) was 5% . 15 cases of heteroplasmic mtDNAs were found by this method in the samples tested. [Conclusion] The established method of PCR-DHPLC can be used for screening the heteroplasmy of mtDNA.更多还原