用新设计引物调查武汉汉族Penta D和Penta E基因座遗传多态性

目的　研究PentaD和PentaE基因座分型引物设计 ,调查PentaD和PentaE基因座在武汉汉族人群中的遗传多态性。方法　用重新设计的PentaD和PentaE基因座分型引物 ,采用热启动PCR和PAGE技术对 2 81名武汉地区汉族无关个体进行分型调查 ,并将其分型结果与Promega公司的PowerPlexTM16系统荧光标记复合扩增试剂盒分型结果进行比较。结果　新设计引物扩增产物的片段大小范围分别为 15 3～ 198bp和 10 7～ 2 12bp ,其分型结果与PowerPlexTM16系统的分型结果完全一致 ,且用银染法检测的灵敏度显著提高 (由 0 5ng提高到 0 2ng)。PentaD和PentaE基因座在武汉汉族群体分别检出 10个和 2 1个等位基因 ,其基因型频率分布均符合Hardy Weinberg平衡。家系调查证实了其等位基因的传递符合孟德尔遗传规律。两基因座的个体识别能力 (DP)分别为 0 92 62、 0 9860 ,非父排除率 (PE)分别为 0 665 1、 0 83 2 5。结论　新设计引物用于PentaD和PentaE基因座的分型检测准确可靠 ,两基因座多态性程度高 ,在法医学个人识别和亲子鉴定中具有使用价值。

Genetic polymorphisms of the pentanucleotide repeat loci Penta D and Penta E in Wuhan Han population by re-designed primers

Objective To study the primer design for Penta D and Penta E genotyping and investigate the genetic polymorphisms of these two loci in Chinese Han population in Wuhan.Methods 281 unrelated Chinese individuals living in Wuhan were typed by hot-start PCR with re-designed Penta D and Penta E genotyping primers and PAGE technique,and the results were compared with those by the PowerPlex TM 16 system of Promega.Results The amplified fragment size of Penta D and Penta E loci with re-designed primers ranged between 153～198bp and 107～212bp respectively,which were consistent with the results obtained by the re-designed primers and PowerPlex TM 16 system,and the re-designed primers had a higher sensitivity than PowerPlex TM 16 system (0.2ng vs 0.5ng) by silver staining.10 and 21 alleles were observed for Penta D and Penta E in Chinese Han population in Wuhan,and the genotype distributions of the two loci were in accordance with Hardy-Weinberg equilibrium.Family studies confirmed Mendelian inheritance of alleles.The power of discrimination (DP) for Penta D and Penta E were 0.926 2 and 0.986 0,and the power of exclusion (PE) were 0.665 1 and 0.832 5 respectively.Conclusion The re-designed primers for Penta D and Penta E genotyping are reliable.These two loci are highly polymorphic in Chinese Han population and can be used in forensic identification and paternity test.