PCR-SSP技术对广东汉族人HLA-DR基因分型

探索具有高分辨率、高特异性和简捷快速的方法对ＨＬＡ－ＤＲ基因分型，为临床器官移植配型和疾病相关性分析提供实用的方法和基础资料．利用ＤＲ１～ＤＲｗ１８序列特异性的１９组引物及１对内参照引物进行ＰＣＲ扩增即ＰＣＲ－ＳＳＰ对ＨＬＡ－ＤＲ进行基因分型，扩增产物经琼脂糖凝胶电泳，溴乙锭染色，在紫外光下观察分型结果．每个被检个体的ＤＲ型别可由特异引物扩增出现的电泳谱带直接判断．双盲检测２２例的结果１００％正确．在１０２例中国广东地区汉族人中，ＤＲ９和ＤＲ２的基因频率最高，分别为０．２２０５和０．１９１２，ＤＲ１０为最低（０．００９８）．与用ＰＣＲ－ＳＳＯ方法分型获得的结果比较，基因型别分布基本一致，但一些等位基因的频率有差异，表明ＨＬＡ－ＤＲ基因频率的分布在不同地区、不同种族的人群间存在着差异．ＰＣＲ－ＳＳＰ法分辨率和特异性虽不及ＰＣＲ－ＳＳＯ法但比血清学方法精细，分型的全过程只需２～４ｈ能满足临床器官移植配型的要求．基因频率调查结果为器官移植配型和疾病相关性分析提供了基础资料．

Genotyping of HLA DR Subergion with PCR SSP in Guangdong Hans

In order to search a functional method and the basic data for transplantation matching and disease correlation analysis,the method of HLA DR genotyping with high resolving power,high specificity,simplicity and convenience was studied.PCR SSP was performed using 19 groups of sequence specific primers directed to various DR1 DRw18 specificities and one pair of positive control primers.After agarose gel electrophoresis of amplified product and stained with ethidium bromide,the results were observed by the ultraviolet ray,The DR genotype of the individual tested can be directly determined by the amplified product associated with the specific primer pairs.The results of 22 cases known as DR genotype by double blind test were correct in 100%.The frequencies of DR9 and DR2 were the highest(0 225 and 0 1912 respectively) and the frequency of DR10 was the lowest (0 0098) in 102 random healthy Guangdong Hans.By comparing the gene frequencies obtained in Guangdong Hans with those obtained in Hunan Hans,northern Chinese and Japanese population,similar distribution of DR subtypes was found and marked difference was seen too.PCR SSP typing has proved to be a rapid and inexpensive genotyping method with high accuracy,high specificity,high reproducibility,simplicity and convenience.The whole process requires 2～4 hours only.It satisfies the requirement of clinical transplantation matching.The results have supplied the basic data for transplantation matching and disease correlation analysis.