APPLICATION OF DNA POLYMORPHISMS TO PERSONAL IDENTIFICATION IN FORENSIC ODONTOLOGY

本研究的目的是建立几种DNA多态性的数据库，并将其应用于法医牙科领域和重大灾难案件中的个人身份识别。D12 S67基因座的差异PCR扩增导致离散等位基因数从11个增加到35个，杂合度从80%增加到93%。比较188个等位基因的核苷酸序列，进一步增加了等位基因的数量为52和杂合性为96%。分别对170、141、155、119和131个样本进行了D12 S66、D21 S11、VWA、DYS 19和D4 S43位点的多态性分析。D12 S66、D21 S1、VWA和D4 S43的杂合度分别为57%、80%、79%和81%，等位基因15在DYS 19的日本人群中占优势。在D21 S11和VWA基因座上发现了与已知等位基因不同的新等位基因，其中等位基因14的核苷酸序列构象与VWA基因座上的其他等位基因不同。D4 S43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、D12S6 D21 S11和VWA基因座在灵长类动物中的分布支持了灵长类动物的分化模式，也显示了STR典型突变模式的某些特征。DNA已经建立。在200摄氏度下热处理的牙齿中可以扩增DNA。在室温下放置2个月的样本中也可以检测唾液染色树脂片中的DNA。我们还从理论上证明了D12 S67、D21 SI 11和VWA这三种多态性可以在超过1000人的大规模灾难中区分个体。

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.