应用mAPLP方法分析湖南汉族、苗族和土家族mtDNA编码区多态性

Detection of mtDNA coding region variants using mAPLP in Han,Miao and Tujia populations from Hunan Province

为建立线粒体DNA编码区SNP快速分型方法,在线粒体DNA编码区选取16个SNP位点(5178A、10398A、14979C、8020A、13104G、11959G、10400T、14178C、3970T、5417A、11969A、12811C、10873T、4580A、7028C、12612G),采用多重扩增产物片段长度多态性分析方法,对湖南地区汉族、苗族和土家族各100人进行了mtDNA编码区多态性分析。结果显示SNP3970T在汉族和土家族人群中的分布频率(均为17%)与苗族相比(8%)存在明显差异(P<0.01),SNP8020A在汉族人群中的分布频率(6%)与苗族和土家族人群(分别为2%和0%)相比存在差异(P<0.05)。在所分析的300名个体中,共检测到45种单倍型,3个民族共有的单倍型12种,两个民族共有的有10种,有23种单倍型仅在1个民族中出现,其中汉族特异性的单倍型有8种,苗族特异性的有6种,土家族特异性的单倍型有9种。mAPLP是通过设计两条不同的正向或反向引物(使PCR扩增片段长度不同)和1条共用的反向或正向引物,使两个等位特异扩增片段大小不同,从而达到SNP分型。

To find a rapid single nucleotide polymorphism （SNP） loci typing method in the mitochondrial DNA （mtDNA） coding regions, we genotyped 16 SNP loci in the mitochondrial DNA coding region in Hart, Miao and Tujia populations by the multiplex-amplified product-length polymorphism （mAPLP） technique. This method generates allele-specific fragments that are different in length through PCR amplification using allele-specific forward （or reverse） primers different in size and a common reverse （or forward） primer. Results showed that both of the allelic frequency of 3970T in Han and Tujia populations were 17%, which were significantly different from that of in the Miao population （P〈0.01）. The allelic frequency of 8020A in Han population was 6%, which was different from that of in the Miao and Tujia populations （P〈0.05）. In all of 300 samples, a total of 45 different haplotypes were identified, 12 of which were found in all the three populations, 10 were shared by two populations and 23 haplotypes existed only in one of the populations. Among these 23 haplotypes, 8, 6 and 12 haplotypes were exclusively observed in the Han, Miao and Tujia populations, respectively.