mtDNA—HVⅠ和细胞色素b片段的复合扩增及其法医学应用

目的探讨复合扩增mtDNA D环HV I和细胞色素b片段进行种属鉴定和个体识别的方法及mtDNA-HV I多态性。方法用两对引物同步扩增HV I片段与细胞色素b片段,银染显带检测扩增产物,ABI377测序仪及荧光测序技术分析扩增产物序列多态性。结果人类有279bp,358bp两条带,动物只有358bp一条带。通过对131例随机广东汉族人群个体进行mtDNA控制区(15997～16236))序列测定统计,得出此区域的序列多态性。共发现69个位点变异,平均每个个体存在2.679个碱基突变,检出67个单倍型,基因多样性为97.92％。结论mtDNA控制区(15997—16236)具有较高的序列多态性。为良好的个体识别标记。复合扩增mtDNA D环HV I与细胞色素b片段进行测序分析可以同步进行种属鉴定和个体识别。

The polymorphism of mtDNA HVI and the application of multiplex amplification of two mitochondrial DNA region to the species and individual identification

Objective To discriminate the species and individual identification with mitochondrial DNA (mtD-NA) sequencing in forensic medicine practice. Methods The multiplex PCR of mtDNA loop - D high - variation region and cytochrome- b region were investigated. The PCR products were detected with silver- stain method,followed by analysis of the PCR products with fluorescence sequence technique. Results The presence of two bands (358bp,279bp ) indicated the samples were from human, while only one band of 358bp indicated nonhuman origin. The part of mitochondrial DNA loop - D high - variation region (15997 ~ 16236) from 131 unrelated individuals of Guangdong population were sequenced. In all of these samples there were 69 nucleotide variations and 67 haplo-types.There was 2.679 mutation sites on average per person. The polymorphism was 97.92% . Conclusion The methods described here are reliable and very useful in species and personal identification of degraded samples.