Study On MtDNA HVI Polymorphism Of Zhenxiong Wang Surname And Forensic Application

[Objective]In order to obtain the mtDNA HVI polymorphism information of the family of Wang surnamed in the villages of Changba Town and Poji Town,Zhenxiong City,the population genetic data was provided for the application of forensic medicine;Optimization of hair stem template DNA extraction,HVI amplification and sequencing to improve the detection rate of such samples.[Method]There are 542 samples of 277 Wang surnames from 16 villages in Zhenxiong County.Those samples were amplified by mtDNA HVI primers and purified by PCR amplification,and the polymorphic information of the sample sequence was statistically analyzed.Statistical analysis was performed using software such as MEGA X and DnaSP5.For the collected hair shaft and nail samples,the modified QIA Investigator kit method and the modified MagAttract M48 DNA Manual kit method were used to extract the mtDNA template.We used the normal PCR and nested PCR amplification samples.At last we Statistics and analysis of polymorphic point information[Result]1.The 542 blood samples's mtDNA HVR-1 wre amplified by nearby sequencesand the sequence length was 550 bp.In this study,we actually detected 298 haplotypes and21 74 mutations occurred in 169 polymorphic points.The polymorphism was mainly converted,accounting for the total polymorphic point.88.30%,of which C/T conversion accounted for 73.39%,and the transversion accounted for 11.19%of the total polymorphism,mainly based on A/C variation.Up to 9 polymorphic loci were detected in a single sample,and at least 0 polymorphic points were detected(consistent with the rCRS sequence).2.The haplotype diversity and nucleic acid diversity of 470 Wang samples were 0.9875±0.002 and 0.01051±0.00024,respectively.3.It is different from the HVI of mtDNA in Zhenxiong Wang and the Han nationality in other regions.4.The PCR products obtained by the silicon membrane method and the magnetic bead method were amplified by ordinary PCR and nested PCR.After agarose electrophoresis,both amplifications were effective to obtain amplified fragments,which were extracted by QIA silicon membrane method.The template DNA can be obtained by ordinary amplification to obtain the target fragment,and the template DNA extracted by the M48 magnetic bead method can also amplify the target fragment after nest amplification.5.The mtDNA HVI sequences of hair shaft and nail samples were statistically analyzed.Among the 24 samples involved,8 haplotypes and 20 polymorphic loci were detected,and 31 bases were converted into polymorphic points.There were 2 sites in the 5th and 6th samples,and no missing sites were detected.6.The sequencing results of hair and nails of the same individual were the same,and no heterogeneity was found.[Conclusion]1.The mtDNA HVI population genetic data of 542 male Wangs from 2 towns of zhenxiong county obtained in this study confirmed that the mtDNA HVR-1 of Zhenxiong County has a rich polymorphic locus;2.There are similar Y chromosome haplotypes in same surname,.We tested mtDNA of males of the same surname to provid a new perspective for further individual identification;3.Direct amplification combined with direct sequencing method can effectively detect the polymorphism of mtDNA HVI,and can provide clues or basis for the investigation of related cases by using higher haplotype diversity;4.The improved QIA silicon membrane method has more advantages in the extraction and amplification of hair stem DNA than the M48 magnetic bead method;5.The areas of mtDNA HVI polymorphism is good,can be used in forensic practice.