用PCR-DGGE技术检测人外周血mtDNA HVⅠ单倍型及异质性

Haplotype and heteroplasmy in the HVⅠ region of human mtDNA detected by PCR-DGGE

目的 建立简单、有效的mtDNA单倍型检测及异质性筛查技术，并获取其相应的汉族人群频率分布。方法 用PCR-DGGE技术对200例武汉汉族无关个体外周血mtDNA HV Ⅰ15997～16174nt和16208—16401nt区域进行分型检测。结果 200例汉族无关个体中，15997～16174nt和16208～1640Int区域分别检出20种和22种单倍型，其单倍型多样性（HD值）分别为0．8159和0．8844；mtDNA HV Ⅰ组合单倍型共90种，其HD值达0．9803。两区域分别有4名和2名个体观察到异质性，其发生率为3％。结论 PCR-DGGE是一种简单、灵敏、高效的mtDNA多态性及异质性检测技术，可应用于法医学实践。

Objective To establish a simple and effective technique for detecting haplotype and heteroplasmy of human mtDNA. Methods Blood samples were collected from 200 unrelated Han individuals in Wuhan. The fragments from 15997 - 16174nt and 16208 - 16401nt of mtDNA HV Ⅰ were analyzed using PCR and denaturing gradient gel electrophoresis （PCR-DGGE）. Results 20 haplotypes were observed from 15997 - 16174nt and the haplotype diversity was 0. 8159, while 22 haplotypes were observed from 16208 -16401nt and the haplotype diversity was 0. 8844. There were 90 haplotypes in combined two fragments from 15997 ～ 16174nt and 16208 ～ 16401nt of mtDNA HV Ⅰ and the haplotype diversity was 0.9803. The heteroplasmy was observed in 15997 - 16174nt of mtDNA from 4 individuals and in 16208 - 16401nt from 2 individuals. Conclusion PCR-DGGE was a simple, sensitive and effective technique for analyzing polymorphism and heteroplasmy of mtDNA.