杂合性歧义引物分离法在解决人类白细胞抗原基因歧义分型结果中的应用

Application of heterozygous ambiguity resolution primers resolving ambiguous genotyping results of human leukocyte antigen genes

目的评估杂合性歧义引物分离法（HARPs）在解决中国汉族人群人类白细胞抗原（H／A）基因歧义分型结果中的应用价值，并筛选合适的HARPs引物。方法416名南方汉族个体的HLA—A、HLA—B、HLA—DRB1基因通过PCR-测序分型（PCR—SBT）方法进行基因分型，并采用美闰Atria公司的HARPs引物对其中的歧义分型标本进行附加测序分型。结果86．3％（132／153）的HLA—A、73．9％（130／176）的HLA-B和38．1％（85／223）的HIM—DRB1歧义分型标本可以通过HARPs方法有效解决；其中48．5％（64／132）的HLA-A、80．0％（104／130）的HLA—B和100．0％（85／85）的HIM—DRB1歧义分型标本只需要1种HAPRs引物，47．7％（63／132）的HLA—A、20．0％（26／130）的HLA-B需要同时使用2种HAPRs引物；3～6种HARPs引物可以解决90％以上的歧义分型标本。结论HARPs可作为解决中国汉族人群HLA—A、HLA—B、HL4-DRB1基因SBT歧义分型结果的常规方法。

Objective To evaluate the heterozygous ambiguity resolution primers （HARPs） method in resolving ambiguous genotyping results of human leukocyte antigen （HLA） genes in Chinese Han population, and choose some appropriate HARPs primers. Methods HLA-A, HLA-B and HLA-DRB1 genes of 416 southern Chinese Han individuals were genotyped by sequence-based-typing（SBT） method and then the ambiguous genotyping samples were sequenced again by HARPs primers provided by American Atria company. Results The percentage of ambiguous genotyping samples resolved by HARPs for HLA-A, HLA-B and HLA-DRB1 locus was 86. 3% （132/153）,73. 9% （130/176） and 38. 1% （85/223） respectively. Among them, 48.5% （64/132）HLA-A, 80. 0% （ 104/130）HLA-B and all HIA-DRB1 （85/85）samples only need one primer, 47.7 % （63/132）HLA-A and 20. 0% （26/130）HLA-B samples need two primers. Three to six different HARPs primers can resolve more than 90% ambiguities. Conclusion HARPs is a convenient method and could be a routine method to resolve ambiguities for HLA-A, HLA-B and HIA-DRB1 genes genotyped by SBT in Chinese Han population.