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的DNA的来源。通常，牙齿DNA是从牙髓获得的。因此获得的DNA通常包含高分子重量DNA，适用于多焦点DNA探针分析。在大多数情况下，在法医实践中看到，样品牙齿在潮湿的条件下留下了，牙髓已被核酸内切酶降解，因此其状况太差，无法分析。我有时发现从整个牙齿截面获得的牙齿DNA严重降解，仅适用于聚合酶链反应（PCR）。此外，在几次情况下，PCR技术也不会放大它，即使琼脂糖凝胶电泳证实了它有足够的DNA来供PCR。结论是，基于CHELEX的程序招标DNA从适合PCR扩增的整个牙齿截面获得，并且使用纯化的牙齿DNA进行法医DNA分析的可能性。传统上，牙科鉴定是基于对末端与邮政牙科牙科牙齿牙齿牙齿图和X射线的比较。如果这是不可能的，则无可争议的是，镀铬高度变量基因座和线粒体DNA测序的有用性是无可争议的，并且应该用作法医牙态实践中的辅助手段。