DNATyper mtDNA-SNP60^(TM)试剂盒在案件中的应用研究  

Application Research of DNATyper mtDNA-SNP60^(TM)Kit in Forensic DNA Casework

作　　者：杨乐 陈滢 吴俞衡 石妍 齐朝阳 孔祥仕 马温华 YANG Le;CHEN Ying;WU Yuheng;SHI Yan;QI Zhaoyang;KONG Xiangshi;MA Wenhua(Forensic Science and Technology Detachment of Harbin Public Security Bureau,Harbin 150010,China;Beijing Tongda Shoucheng Institute of Forensic Science,Beijing 100192,China;Fangshan Branch of Beijing Public Security Bureau,Beijing 102488,China;Institute of Forensic Science,Ministry of Public Security,Beijing Engineering Research Center of Crime Scene Evidence Examination,Beijing 100038,China;Key Laboratory for Ecotourism of Hunan Province,School of Tourism,Jishou University,Zhangjiajie 427000,Hunan,China)

机构地区：[1]哈尔滨市公安局刑事技术支队,哈尔滨150010 [2]北京通达首诚司法鉴定所,北京100192 [3]北京市公安局房山分局,北京102488 [4]公安部鉴定中心,北京市现场物证检验工程技术研究中心,北京100038 [5]吉首大学旅游学院生态旅游湖南省重点实验室,湖南张家界427000

出　　处：《刑事技术》2024年第3期255-261,共7页Forensic Science and Technology

基　　金：中央级公益性科研院所基本科研业务费专项资金项目(2020JB006);公安部鉴定中心开放课题(2020CSEEKFKT03)。

摘　　要：本文探讨DNATyper mtDNA-SNP60^(TM)试剂盒在案件中应用的可行性。应用DNATyper mtDNASNP60^(TM)试剂盒对100个汉族无关个体和20组全同胞进行mtDNA SNP检验;取25 pg/μL马、牛、羊、猪、鸡、鸭、猫、狗、兔、鼠和大肠杆菌的DNA样品进行种属特异性测试;取5、10、20、40μmol/L血红素进行抗抑制性测试;取两个批次的DNATyper mtDNA-SNP60^(TM)试剂盒经反复冻融10次后进行稳定性测试;分别应用VeriFiler^(TM)Plus PCR扩增试剂盒和DNATyper mtDNA-SNP60^(TM)试剂盒对100份陈旧、腐败、降解检材进行检验。结果表明,100个汉族无关个体均获得清晰的mtDNA SNP分型结果,其检验结果与通过mtDNA测序获得的结果完全一致;100个汉族无关个体含有100种不同的单倍型;20组全同胞中每组个体之间mtDNA SNP分型结果相同;DNATyper mtDNA-SNP60^(TM)试剂盒对马、牛、羊、猪、鸡、鸭、猫、狗、兔、鼠和大肠杆菌的DNA样品进行检测,均未出现特异性分型;当血红素浓度≤40μmol/L时,所有mtDNA SNP位点均获得正确分型;两个批次的DNATyper mtDNA-SNP60^(TM)试剂盒经反复冻融10次后,所有mtDNA SNP位点均可正确分型;对于100份陈旧、腐败、降解检材,STR检出率为55%,mtDNA SNP的检出率为86%,mtDNA SNP的检出率显著高于STR。当模板DNA浓度大于5 pg/μL时,DNATyper mtDNA-SNP60^(TM)试剂盒能得到完整的分型谱图。综上,DNATyper mtDNA-SNP60^(TM)试剂盒可应用于陈旧、腐败、降解检材的检验,具有很好的实战应用价值。The DNATyper mtDNA-SNP60^(TM)kit is a mtDNA SNP multiplex amplification system,which is suitable for the Chinese population.At present,the DNATyper mtDNA-SNP60^(TM)kit has not been officially launched on the market.There has been no systematic research on the application of the DNATyper mtDNA-SNP60^(TM)kit in practical cases.To explore the feasibility of the application of DNATyper mtDNA-SNP60^(TM)kit in forensic DNA casework,DNATyper mtDNA-SNP60^(TM)kit was used to test the mtDNA SNP of 100 unrelated individuals of Han nationality and 20 groups of whole siblings.25 pg/μL DNA samples from horses,cows,sheep,pigs,chickens,ducks,cats,dogs,rabbits,mice,and Escherichia coli were taken to perform species specific testing.5,10,20,40μmol/L heme were taken for anti inhibitory testing.Two batches of DNATyper mtDNA-SNP60^(TM)reagent kits were performed stability testing after repeated freeze-thaw for 10 times.VeriFiler^(TM)Plus PCR amplification kit and DNATyper mtDNA-SNP60^(TM)kit were used to detect the 100 old,rotten and degraded samples respectively.Results showed that all 100 unrelated individuals of Han nationality had clear mtDNA SNP typing,and the results were completely consistent with those obtained by mtDNA sequencing.Under the detection of DNAtyper mtDNASNP60^(TM)kit,the 100 unrelated individuals of Han nationality showed 100 different haplotypes,and the mtDNA SNP typing was the same among individuals in each group of the 20 whole siblings’groups,which suggested that the DNATyper mtDNASNP60^(TM)kit can be used for maternal lineage identification.DNATyper mtDNA-SNP60^(TM)kit was used to detect DNA samples from horses,cows,sheep,pigs,chickens,ducks,cats,dogs,rabbits,mice,and Escherichia coli,but no specific typing was found,indicating that the kit has good species specificity.When heme concentration at≤40μmol/L,all mtDNA SNP sites were correctly typed,indicating that the kit has certain anti heme ability.Two batches of DNATyper mtDNA-SNP60^(TM)reagent kits were subjected to repeated freeze-thaw for 10 times

关 键 词：法医遗传学 DNATyper mtDNA-SNP60^(TM)试剂盒 线粒体DNA 单核苷酸多态性 VeriFiler^(TM)Plus PCR扩增试剂盒 短串联重复序列 

分 类 号：D919[医药卫生—法医学]