HLA—A基因座分步PCR—SSP分型研究

目的使HLA基因分型能应用于法医常见检材的个人识别。方法 建立检测HLA—A基因座的分步PCR—SSP方法。先用一对HLA—A基因座特异的引物作第一次扩增,以所得产物为模板,分别用对HLA—A30、A31、A33特异的3对引物作第二次扩增,二次扩增的产物经电泳判型。结果 1130例血清分型为HLA—A30、A31、A33的血痕,其PCR—SSP分型和血清分型的不符合率为29％;室温保存2年的精斑、唾液斑,保存18年的血痕第一次扩增均获得满意的结果。结论法医亲子鉴定和个人识别宜用基因分型替代血清分型。HLA—A基因座分步PCR—SSP基因分型适用于法医检材。

Two - step PCR - SSP genotyping for HLA - A locus and its comparison with serological typing results

Objective To identify routine forensic samples by genotyping method for HLA - A locus. Method A two - step PCR - SSP method was established. The first step is amplification with a pair of primers specific for all HLA - A alleles, the second step is amplification with primers specific for HLA - A30, A31, A33 respectively, using the first step amplification product as template. Secondly amplified PCR products were genotyped by electro-phoresis. Results 100 blood stains with a serological typing result of HLA - A30, A31 and A33 were tested with this method. The discrepancy rate between serological and genetic typing was 29%. Seminal stains, salivary stains reserved for 2 years and blood stains reserved for 18 years in room temperature gave satisfactory results. Conclusions It is better to replace serological typing by genetic typing. A two - step PCR - SSP genotyping method can be applied to forensic samples.