The study of HLA-DNA typing technique applicable to forensic investigation

我研究了基于聚合酶链式反应技术的人类白细胞抗原-DNA配型在法医调查中的适用性。首先，将人类白细胞抗原II类分型与聚合酶链式反应-单链构象多态性和聚合酶链式反应-限制性片段长度多态性相结合应用于法医实践。聚合酶链式反应-单链构象多态配型与血清学HLA-DR配型一样，是确定DR分型的有效方法。采用聚合酶链式反应-限制性片段长度多态性方法对DRB1编码DR2、DDR4和DR8的等位基因进行更精确的定义。此外，我还使用了套式或半套式聚合酶链式反应技术来提高该体系的灵敏度。其次，研究了用聚合酶链式反应-单链构象多态方法进行人类白细胞抗原I类分型的适用性。我设计了96种混合引物，用来扩增与血清学I类抗原相对应的人类白细胞抗原-A、-B、-C等位基因，这些方法可以从法医样本的DNA中扩增出人类白细胞抗原-DNA，但DNA的质量和数量是成功的关键。

I investigated the applicability of HLA -DNA typing using PCR-based technique to forensic investigations. First, I applied the HLA-class II typing combined With PCR-SSP and PCR-RFLP to forensic practice. PCR-SSP typing was an effective as serological HLA-DR typing in determining DR types. PCR-RFLP typing was used for more precise definition of DRB1 alleles encoding DR2, DDR4, and DR8. Furthermore I used the nested or semi-nested PCR technique to increase the sensitivity of this system. Second, I studied the applicability of HLA-class I typing using PCR-SSP method. I made 96 kinds of primer mixture to amplify HLA-A,-B and-C alleles corresponding to serological class I antigens.HLA-DNA typing by these methods were possible from DNA samples extracted from forensic samples, but both quality and quantity of DNA were important for successful.