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DNA analysis of HLA class II gene and application of forensic specimens

HLA II类基因DNA分析及法医标本应用

基本信息

批准号：
09670452
负责人：
YANAGIDA Junichi
金额：
$ 1.86万
依托单位：
Keio University
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 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670452/
关键词：
HLA class II gene DNA polymorphism paternity testing direct PCR

项目摘要

1) HLA DQA 1 and HLA DRB genotypings by the PCR-SSP and PCR-SSO methods were applied to the cases of disputed paternity. Conventional blood group testing and HLA class II genotyping (HLA DQA 1 and HLA DRB) were good agreement with the caes of paternity test.2) In field of forensic medicine, biological samples such as bloodstain, semen, and hair are very important samples for personal identification. It has to spend much time and make many effects to extract DNA from torensic samples. The Drop PCR Kit for human blood, by which PCR reaction directly carried out on whole blood and bloodstain without DNA extraction process, was applied to the analyses of HLA DQα and HLA DRB geontypings. The experimental results suggested that HLA class II genotyping from small amount or bloodstain (size: 1mm x 1mm) by direct PCR could be applied to forensic practices.3) The DNA extracted from the nails of human living bodies and cadavers was examined to clarify its usefulness in the practical analysis of DNA polymorphisms. No significant differences were shown in the recovery and the purity of five different methods (phenol-chloroform) method, SMITEST DNA extraction kit, Wako DNA Extractor WB Kit, DnaQuick and SepaGene) for DNA extraction from nails of living bodies. The DNA from cadaveric nails was able to be extracted by phenol-chloroform method. The applicability of the human nail DNA for the polymorphism analyses with HLA DQ α typing, HLA-DRB typing and PM-PCR typing suggested that the DNA extracted from human nails (both living bodies' and cadavers') is useful the practical test in forensic science.

1)应用PCR-SSP法和PCR-SSO法对父权争议病例进行HLA DQA 1和HLA DRB基因分型。常规血型检测和HLAⅱ类基因分型（HLA DQA 1和HLA DRB）与亲子鉴定结果吻合较好。2)在法医学领域，血迹、精液、毛发等生物样本是非常重要的个人鉴定样本。从化石样本中提取DNA需要花费大量的时间和很多的方法。人血Drop PCR试剂盒，直接在全血和血迹上进行PCR反应，无需提取DNA，用于HLA DQα和HLA DRB的分型分析。实验结果表明，从少量或1mm × 1mm的血迹中直接PCR分型HLAⅱ类基因可应用于法医实践。3)对从人体和尸体指甲中提取的DNA进行检验，以阐明其在DNA多态性的实际分析中的实用性。5种不同方法（苯酚-氯仿法、SMITEST DNA提取试剂盒、Wako DNA提取试剂盒、DnaQuick和SepaGene）对活体指甲DNA的提取回收率和纯度均无显著差异。用苯酚-氯仿法从尸体指甲中提取DNA。人指甲DNA对HLA DQ α分型、HLA- drb分型和PM-PCR分型的多态性分析的适用性表明，从人指甲（无论是活体指甲还是尸体指甲）中提取的DNA可用于法医学的实际检验。

项目成果

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Sugiura,K., Shinozuka, T., Takei, S., Takada,T., Nakajima, R., Ro,A., Tomita,A., Murai,T., Yanagida,J., Hara,M. and Watanabe,H.: "DNA typing or blood stain specimens by PCR method with AmpdirectィイD1TMィエD1 reagent."DNA polymorphism. 6. 72-75 (1998)

Sugiura,K.、Shinozuka, T.、Takei, S.、Takada,T.、Nakajima, R.、Ro,A.、Tomita,A.、Murai,T.、Yanagida,J.、Hara,M 和Watanabe, H.：“使用 Ampdirect D1 试剂通过 PCR 方法进行 DNA 分型或血染标本。”DNA 多态性。 6. 72-75 (1998)