不同接触DNA检材提取纯化方法在法医学中的应用价值比较
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摘要
比较不同DNA检材的提纯方法,便于法医的实际应用选择。采集饮用过的饮料瓶或矿泉水瓶、废弃的烟蒂、使用过的手套口罩及衣物各90份,随机分为Chelex组、磁珠组和有机组,各组检材分别使用相应的方法进行DNA的提纯,对提纯后三组检材的DNA提取液的STR分型、DNA提取量与紫外线分光光度计下OD值水平进行比较。由磁珠法提纯得到的DNA超过9个基因座的例数最多;提取量与DNA提取纯度显著优于其他二组,差异具有统计学意义(p<0.05)。对于微量接触或污染严重的检材,可使用DNA IQ磁珠法进行提纯,可优化DNA的提取纯度,利于法医工作的进行。
To compare the different purification methods of DNA examination,facilitate the forensic application of choice,we collevbed 90 copies each drinking bottles or mineral water bottles,discarded cigarette butts and used gloves,masks,clothes.Divided into Chelex-100 group,immunomagnetic beads group and organic group.Each group using the appropriate method for the purification of DNA,Comparing with the three groups of detection DNA extract STR parting,DNA extraction quantity and uv spectrophotometer OD value level after purification.By the method of immunomagnetic beads purification of DNA over 9 loci most cases;extraction and purity of DNA extraction significantly better than the other two groups.The micro contact or serious pollution for examination,the DNA IQ method of magnetic beads can be used for purification,to optimize the extraction of DNA purity,and conducive to forensic work.
To compare the different purification methods of DNA examination,facilitate the forensic application of choice,we collevbed 90 copies each drinking bottles or mineral water bottles,discarded cigarette butts and used gloves,masks,clothes.Divided into Chelex-100 group,immunomagnetic beads group and organic group.Each group using the appropriate method for the purification of DNA,Comparing with the three groups of detection DNA extract STR parting,DNA extraction quantity and uv spectrophotometer OD value level after purification.By the method of immunomagnetic beads purification of DNA over 9 loci most cases;extraction and purity of DNA extraction significantly better than the other two groups.The micro contact or serious pollution for examination,the DNA IQ method of magnetic beads can be used for purification,to optimize the extraction of DNA purity,and conducive to forensic work.
引文
De Bruin K.G.,Verheij S.M.,Veenhoven M.,and Sijen T.,2012Comparison of stubbing and the double swab method fo collecting offender epithelial material from a victim's skin Forensic Science International:Genetics,6(2):219-223
Dong H.,Wang J.,Li C.X.,Zhang T.,and Liu C.,2015,An ex ploration on trace touch related DNA materials from differ ent substrates,Shengming Kexue Yanjiu(Life Science Res earch),19(2):145-153(董会,王晶,李彩霞,张涛,刘超,2015不同载体上微量接触类检材的相关问题探究,生命科学研究,19(2):145-153)
Feng Y.,Yan M.,Liu Y.,and Chen J.,2008,A method for DNAexamination on mouth mucosa exfoliative cells from toothbrush bristles,Fayixue Zazhi(Journal of Forensic Medlicine)24(6):435-436(封宇,严密,刘云,陈骏,2008,牙刷刷毛黏附口腔黏膜脱落细胞DNA的检验方法,法医学杂志,24(6):435-436)
Kuan J.Z.,Nie T.G.,Yang Z.,Zhu W.,Wang Y.J.,Sun R.,and Ma Y.,2009,A study of automation workstation for th DNA extraction of forensic biological evidence,Zhongguo Fayixue Zazhi(Chinese Journal of Forensic Medicine),24(3):164-167(匡金枝,聂同钢,杨智,朱巍,王玉健,孙睿马妍,2009,法医物证DNA自动化检验技术体系的研究中国法医学杂志,24(3):164-167)
Liu H.B.,Feng B.Q.,Huang Y.,and Xu B.Y.,2012,Comparison of three methods for extracting DNA from shedding of epi dermal cells in contact material,Kunming Yikedaxue Xue bao(Journal of Kunming Medical University),33(12):3640(刘海渤,冯保强,黄颖,许冰莹,2012,3种提取接触性检材脱落表皮细胞DNA方法的实验研究,昆明医科大学学报,33(12):36-40)
Luo J.H.,Chen X.J.,Sun H.B.,Yang X.,Zhang Z.L.,and Wang T.,2015,Comparison of DNA extraction methods from bio logical contact samples,Lanzhoudaxue Xuebao(Yixue Ban(Journal of Lanzhou University(Medical Sciences)),41(1)18-22(骆继怀,陈晓军,孙红兵,杨鑫,张子龙,王涛,2015接触性生物检材提取DNA方法的比较,兰州大学学报(医学版),41(1):18-22)
Mo Y.N.,Xue X.Q.,and Liu L.,2008,Comparison of three DNAextraction methods for genotyping of difficulty mixed stains Zhongguo Fayixue Zazhi(Chinese Journal of Forensic Medicine),23(1):26-28(莫耀南,薛小琦,刘良,2008,3种DNA提取法在污染严重混合斑分型中的应用比较,中国法医学杂志,23(1):26-28)
Qian Z.L.,Li M.,Xiao Q.Q.,Miao C.Y.,Li T.,Zhuang K.Y.,and Guo Z.J.,2016,CABYR biological function and its applica tion,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(6):1335-1341(钱尊磊,李敏,肖倩倩苗翠英,李同,庄柯彧,郭子嘉,2016,CABYR生物学功能及其应用,基因组学与应用生物学,35(6):1335-1341)
Walsh P.S.,Metzgar D.A.,and Higuehi R.,1991,Chelex-100 a a medium for simple extraction of DNA for PCR-based typ ing from forensic material,Biotechniques,10(4):506-513
Yang D.,Liu C.,Xu Q.Y.,and Chen X.H.,2011,Extraction o DNA from“touch”DNA samples by DNA IQ Casewor Sample Kit in MaxwellTM16,Xingshi Jishu(Forensic Sci ence and Technology),(3):3-5(杨电,刘超,徐曲毅,陈向红,2011,DNA IQ磁珠法结合MaxwellT M 16自动仪提取接触DNA,刑事技术,(3):3-5)
Yang D.,Zhang L.P.,Liu C.,and Xu Q.Y.,2012,The compari son of the extraction effects of touch DNA by the Chelex-100 and magnetic beads,Xingshi Jishu(Forensic Science and Technology),1(1):11-13(杨电,张丽萍,刘超,徐曲毅2012,Chelex法和两种磁珠法提取接触DNA效果的比较,刑事技术,1(1):11-13)
Dong H.,Wang J.,Li C.X.,Zhang T.,and Liu C.,2015,An ex ploration on trace touch related DNA materials from differ ent substrates,Shengming Kexue Yanjiu(Life Science Res earch),19(2):145-153(董会,王晶,李彩霞,张涛,刘超,2015不同载体上微量接触类检材的相关问题探究,生命科学研究,19(2):145-153)
Feng Y.,Yan M.,Liu Y.,and Chen J.,2008,A method for DNAexamination on mouth mucosa exfoliative cells from toothbrush bristles,Fayixue Zazhi(Journal of Forensic Medlicine)24(6):435-436(封宇,严密,刘云,陈骏,2008,牙刷刷毛黏附口腔黏膜脱落细胞DNA的检验方法,法医学杂志,24(6):435-436)
Kuan J.Z.,Nie T.G.,Yang Z.,Zhu W.,Wang Y.J.,Sun R.,and Ma Y.,2009,A study of automation workstation for th DNA extraction of forensic biological evidence,Zhongguo Fayixue Zazhi(Chinese Journal of Forensic Medicine),24(3):164-167(匡金枝,聂同钢,杨智,朱巍,王玉健,孙睿马妍,2009,法医物证DNA自动化检验技术体系的研究中国法医学杂志,24(3):164-167)
Liu H.B.,Feng B.Q.,Huang Y.,and Xu B.Y.,2012,Comparison of three methods for extracting DNA from shedding of epi dermal cells in contact material,Kunming Yikedaxue Xue bao(Journal of Kunming Medical University),33(12):3640(刘海渤,冯保强,黄颖,许冰莹,2012,3种提取接触性检材脱落表皮细胞DNA方法的实验研究,昆明医科大学学报,33(12):36-40)
Luo J.H.,Chen X.J.,Sun H.B.,Yang X.,Zhang Z.L.,and Wang T.,2015,Comparison of DNA extraction methods from bio logical contact samples,Lanzhoudaxue Xuebao(Yixue Ban(Journal of Lanzhou University(Medical Sciences)),41(1)18-22(骆继怀,陈晓军,孙红兵,杨鑫,张子龙,王涛,2015接触性生物检材提取DNA方法的比较,兰州大学学报(医学版),41(1):18-22)
Mo Y.N.,Xue X.Q.,and Liu L.,2008,Comparison of three DNAextraction methods for genotyping of difficulty mixed stains Zhongguo Fayixue Zazhi(Chinese Journal of Forensic Medicine),23(1):26-28(莫耀南,薛小琦,刘良,2008,3种DNA提取法在污染严重混合斑分型中的应用比较,中国法医学杂志,23(1):26-28)
Qian Z.L.,Li M.,Xiao Q.Q.,Miao C.Y.,Li T.,Zhuang K.Y.,and Guo Z.J.,2016,CABYR biological function and its applica tion,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(6):1335-1341(钱尊磊,李敏,肖倩倩苗翠英,李同,庄柯彧,郭子嘉,2016,CABYR生物学功能及其应用,基因组学与应用生物学,35(6):1335-1341)
Walsh P.S.,Metzgar D.A.,and Higuehi R.,1991,Chelex-100 a a medium for simple extraction of DNA for PCR-based typ ing from forensic material,Biotechniques,10(4):506-513
Yang D.,Liu C.,Xu Q.Y.,and Chen X.H.,2011,Extraction o DNA from“touch”DNA samples by DNA IQ Casewor Sample Kit in MaxwellTM16,Xingshi Jishu(Forensic Sci ence and Technology),(3):3-5(杨电,刘超,徐曲毅,陈向红,2011,DNA IQ磁珠法结合MaxwellT M 16自动仪提取接触DNA,刑事技术,(3):3-5)
Yang D.,Zhang L.P.,Liu C.,and Xu Q.Y.,2012,The compari son of the extraction effects of touch DNA by the Chelex-100 and magnetic beads,Xingshi Jishu(Forensic Science and Technology),1(1):11-13(杨电,张丽萍,刘超,徐曲毅2012,Chelex法和两种磁珠法提取接触DNA效果的比较,刑事技术,1(1):11-13)