用聚合酶链反应和生物素寡核苷酸探针分析HLA-DQα的基因型

Analysis of HLA-DQα Genotypes by Amplified DNA Fragments Probed With Biotinylated Oligonucleotides

来自不同人的染色体DNA经聚合酶链反应(PCR)将其HLA-DQα基因的多态区242bp人工地扩增30个循环。用1/10扩增产物点在尼龙膜上。将四种不同的等位基因特异的寡核苷酸(ASOs)经Bio-11-duTP进行3'末端标记得生物素探针。以此探针对不同扩增标本做狭缝印迹杂交。结果表明用此种非放射性的方法能探测出人白细胞表面抗原DQα基因的微多态性,从而得到不同个体的基因型。ASO分型是对血清HLA分型的重要补充和发展,它在骨髓或器官移植以及法医的个人识别中有一定实用价值。

HLA-D/DR matching was quite important in BMT and organ transplantation in connexion with the protection of GVHD. Recently HLA-D oligonucleotide typing has been developed. In this paper we introduced PCR and biotin-labelling technics to examine the HLA-DQα genotype of individuals. This nonradioactive technic could be applied in forensic medicine as well.