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Extraction Method of Dental DNA by Using whole Tooth Sections : Application to Forensic Odontology

使用全牙切片提取牙齿 DNA 的方法：在法医牙医学中的应用

基本信息

批准号：
08672143
负责人：
YAMADA Yoshihiro
金额：
$ 0.9万
依托单位：
Kanagawa 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：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08672143/
关键词：
forensic odontology personal identification mitochondrial DNA mass disaster DNA purification method chelex ion exchange resin DNA精製法歯科法医学法医DNA分析 PCR阻害物質 Chelex

项目摘要

I have previously reported the forensic value of teeth as a source of DNA for genetic analysis. Commonly, dental DNA has been obtained from dental pulp. The DNA thus obtained usually contains high-molecular-weight DNA and is suitable for multilocus DNA probe analysis. In most cases seen in forensic practice, sample teeth have been left in humid conditions and the dental pulp has been degraded by endonucleases, so that its condition is too poor for analysis. I have sometimes found that dental DNA obtained from a whole tooth section is badly degraded, and suitable only for polymerase chain reaction (PCR). Moreover, on several occasions, it is not amplified by the PCR technique even though it has been confirmed by agarose gel electrophoresis that there is enough DNA for the PCR.Therefore, I suspected that some cations contaminated in tooth sections might contaminate dental DNA during its extraction as a source material for PCR,and act as a inhibitor of Taq DNA polymerase during the PCR procedure.In this study, I demonstrated conclusively that the Chelex-based procedure tenders DNAs obtained from a whole tooth section suitable for PCR amplification, and the possibility of forensic DNA analysis using those of purified dental DNA.Dental identification is traditionally based on a comparison of antemortem with postmortem dental charts and X-rays. In cases where this is impossible the usefulness of the amplification of chromosal hypervariable loci and mitochondrial DNA sequencing are undisputed and it should use as an adjunct in forensic odontological practice.

我以前曾报道过牙齿作为遗传分析DNA来源的法医价值。通常，牙齿DNA已经从牙髓中获得。这样获得的DNA通常含有高分子量DNA，适合于多位点DNA探针分析。在法医实践中看到的大多数情况下，样本牙齿被放在潮湿的条件下，牙髓已被核酸内切酶降解，因此其状况太差，无法进行分析。我有时发现从整个牙齿切片中获得的牙齿DNA严重降解，只适合聚合酶链反应（PCR）。此外，有几次，即使琼脂糖凝胶电泳证实有足够的DNA用于PCR，PCR技术也不能扩增。因此，我怀疑牙齿切片中污染的某些阳离子可能在提取牙齿DNA作为PCR源材料时污染牙齿DNA，并在PCR过程中作为Taq DNA聚合酶的抑制剂。在本研究中，我证明了结论性的Chelex为基础的程序招标DNA从整个牙齿部分适合PCR扩增，和法医DNA分析的可能性，使用那些纯化的dental DNA.Dental识别传统上是基于比较生前与死后的牙科图表和X射线。在无法做到这一点的情况下，染色体高变基因座扩增和线粒体DNA测序的有用性是无可争议的，它应该在法医牙科实践中作为一种辅助手段。