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

1)对亲子鉴定有争议的病例，分别采用聚合酶链式反应-单链构象多态和聚合酶链式反应-单链构象多态方法进行HLADQA1和HLADRB基因分型。2)在法医学领域，血迹、精液、毛发等生物样本是进行个人身份鉴定的重要样本。从法医样本中提取DNA需要花费大量的时间和产生很多效果。应用Drop-PCRKit直接对全血和血痕进行聚合酶链式反应，不需要提取DNA，用于HLADQα和HLADRB基因分型。实验结果表明，从微量或血迹(大小：1 mm×1 mm)中提取的DNA可以应用于法医实践。3)对从活体和身体指甲中提取的DNA进行检测，以阐明其在DNA多态性分析中的实用性。从活体指甲中提取DNA的5种方法(苯酚-氯仿法、SMITEST DNA提取试剂盒、Wako DNA Extractor WB试剂盒、Dna Quick和Sepagene)的回收率和纯度均无显著差异。用苯酚-氯仿法可以从身体指甲中提取DNA。人指甲DNA在HLADQDRB型、HLADRB型和PM-α分型中的适用性表明，从活体和身体指甲中提取的DNA可用于法医学的实用检验。

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.