生物素掺入反向杂交法在法医学中应用的研究

Forensic application of reverse dot blot with biotin incorperation

建立了一套快速、准确检测人类 HLA－ DQA基因的反向杂交检测技术，可准确判定 HLA－ DQA基因位点的 6个等位基因即 0101、 0102、 0103、 0201、 0301、 0401。调查了中国北方汉族人群中 200例无关个体的 HLA－ DQA基因频率及基因型的频率分布，杂合度 (H)为 0.75,个体识别能力 (DP)为 0.928。采用生物素直接掺入 PCR扩增的方法， 1ng的 DNA样品即可准确判型。对血液、血斑、精斑、精液与阴道液的混合斑、肌肉组织等检材进行检测，效果良好。在刑事案件及亲子关系鉴定中应用，准确地判定了检材的 HLA－ DQA基因型，为侦察破案提供了科学依据。将此项技术商品化，完成了试剂盒的研制。

A rapid and accurate reverse dot blot technique was successfully established. It could distinguish the 6 alleles of 0101, 0102, 0103, 0201, 0301 and 0401 of HLA－ DQA locus. Alleles and genotype frequencies for HLA－ DQA locus were determined in 200 unrelated individuals of northern Han population. The discrimination power (DP) was 0.928, heterozygocity (H) 0.75. By direct biotin incorperation, 1ng genome DNA could be detected successfully. This method can be used for paternity test and individual identification in forensic science practice.