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APPLICATION OF DNA POLYMORPHISMS TO PERSONAL IDENTIFICATION IN FORENSIC ODONTOLOGY

DNA多态性在法医齿科个人身份识别中的应用

基本信息

批准号：
09672120
负责人：
MINAGUCHI Kiyoshi
金额：
$ 2.18万
依托单位：
TOKYO DENTAL COLLEGE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

The purpose of this study is preparation of database on several DNA polymorphisms and their application to personal identification in the field of forensic dentistry and in the case of mass disaster. Differential PCR amplification of D12S67 locus resulted in increase in the number of discrete alleles from 11 to 35 and increase of heterozygosity from 80% to 93%. Comparison of nucleotide sequences of 188 alleles further increased the number of alleles to 52 and heterozygosity to 96%. Polymorphisms of D12S66, D21S11, VWA, DYS 19, and D4S43 loci were investigated using 170, 141, 155, 119 and 131 samples, respectively. The heterozygosity calculated for D12S66, D21S1, VWA and D4S43 were 57, 80, 79 and 81%, respectively.The frequency of allele 15 was dominated in the Japanese population in DYS 19. New alleles with different nucleotide sequences from the known alleles were found in D21S11 and VWA loci, and the conformation of the nucleotide sequence of allele 14 was different from other alleles in the VWA locus. Allele 1 of the D4S43 locus was further classified into 4 alleles, resulting in the increase of heterozygosity of the locus. Above mentioned loci could successfully applied to the detection of polymorphisms using teeth as a source of DNA.Comparison of the nucleotide sequences of D12S67, D21S11 and VWA loci in primates supported the known divergence models of primates and also showed some characteristics of typical mutation pattern of STR.Unknown regions of the X and Y amelogeniri genes were sequenced and new methods of sex determination using highly degraded DNA was established. Amplification of DNA was possible from teeth heat-treated at 200ﾟC.Detection of DNA from saliva-stained resin pieces was also possible from samples left at room temperature until 2 months. We also showed theoretically that three polymorphisms, D12S67, D21SI11 and VWA, could discriminate individuals in a mass disaster with victims of more than 1000.

本研究的目的是建立几种DNA多态的数据库，并将其应用于法医牙科领域和大规模灾难案件中的个人身份识别。D12S67基因座的差异PCR扩增使离散等位基因数从11个增加到35个，杂合度从80%增加到93%。188个等位基因的核苷酸序列比较进一步使等位基因数目增加到52个，杂合度增加到96%。分别对170、141、155、119和131个样本进行了D12S66、D21S11、VWA、DYS19和D4S43基因座的遗传多态性分析。D12S66、D21S1、VWA和D4S43的杂合度分别为57、80、79和81%。等位基因15在DYS19的日本群体中占主导地位。在D21S11和VWA座位上发现了与已知等位基因核苷酸序列不同的新等位基因，等位基因14的核苷酸序列构象与VWA座位上的其他等位基因不同。D4S43基因座的等位基因1进一步分为4个等位基因，导致该基因座的杂合度增加。D12S67、D21S11和VWA基因座在灵长类动物中的核苷酸序列比较支持灵长类动物的已知分化模式，也显示了STR典型突变模式的一些特征。对X和Y基因的未知区域进行了测序，建立了利用高度降解的DNA进行性别鉴定的新方法。从200ﾟC热处理的牙齿中扩增DNA是可能的，从唾液染色的树脂片中检测DNA也是可能的，从室温下保留2个月的样本中也可以检测到DNA。我们还从理论上证明了D12S67、D21SI11和VWA这三个基因多态可以将大规模灾难中的个体与1000多名受害者区分开来。