云南汉族血小板献血者HLA-A、B和HPA基因多态性的分析

Gene Frequencies of the HLA-A,B and HPA in Typed Platelet Donors of Yunnan Han Nationality

目的研究云南地区汉族血小板献血者HLA-A、B基因和HPA1-17基因多态性,建立已知型血小板献血者基因数据库。方法分别采用PCR-SSOP和PCR-SSP方法对1 000名云南地区无血缘关系的汉族血小板献血者做HLA-A、B和HPA1-17系统基因分型,计算基因型频率、基因频率,并与其他人群进行比较。结果在云南地区人群中共检出HLA-A位点14个,HLA-B位点35个。A*02(35.81%)、A*11(35.42%)和A*24(17.72%)3个等位基因为云南汉族人群HLA-A座位的主要等位基因;B*46(12.87%)、B*40(60)(10.83%)和B*15(62)(9.78%)3个等位基因为云南汉族人群HLA-B座位的主要等位基因。每个样本均检测到HPA-1a、2a、4a-14a、16a、17a基因;HPA-4a、7a-14a、16a、17a呈现单态性,未检测出相应的等位基因HPA-b;对于HPA-1、2、5、6主要以a/a纯合子居多,a/a基因型频率分别是0.989、0.960、0.990、0.980,没有b/b纯合子出现。在HPA1-17中,具有最高杂合度的是HPA-15,基因型HPA15a/15a、HPA15a/15b、HPA15b/15b频率分别是0.392、0.360、0.248;HPA-3在其次,基因型HPA3a/3a、HPA3a/3b、HPA3b/3b频率分别是0.339、0.467、0.194。经χ2检验,结果符合Hardy-Weinberg遗传平衡定律。结论云南地区汉族血小板献血者HLA-A、B和HPA1-17基因频率与南方汉族接近,也呈现其自身特点。应建立本地区的血小板供者分型数据库。

Objective To investigate the genetic polymorphisms of HLA-A, B alleles and HPAI-17 in Yunnan Han population platelet donors and to establish typed platelet donor data bank. Methods The technique of flow polymerase chain reaction sequence specific oligonucleotide probes （PCR-SSOP） and PCR-sequence specific primers（PCR-SSP） meth- ods were respectively adopted in genotyping HLA-A,B and HPAI-17. Samples of 1 000 unrelated platelet dornors were collected from Kunming,Yunnan province, China. The allele frequencies of HLA-A, B and HPAI-17 were calculated, and the allele frequency distribution in this population was compared with the results from various regions. Results The num- ber of alleles identified was 14 for HLA-A, 35 for HLA-B, respectively. The most frequent HLA-A alleles were A＊ 02 （35.81%）, A＊ 11 （35.42% ） and A＊ 24 （17.72 % ）. For the HLA-B locus, the most common in descending order were B＊ 46 （12.87%） ,B＊ 40（60）（10.83%） and B＊ 15（62）（9.78%）. The HPA results indicated that HPA -1a, 2a, 4a-14a, 16a and 17a genes were found among the 17 HPAs in every sample tested. Monomorphic HPA-4a, 7a-14a, 16a and 17a were found in the samples. For HPA-1, 2, 5 and 6, a/a homozygosity was predominant with fequencies of 0. 989,0. 960,0. 990,0. 980 respectively, and none of HPA b/b was found in the samples. HPA-lb, 2b, 5b, 6b were rarely found among subjects. HPA-15 had the greatest heterozygosity with a gene frequency of 0. 392,0. 360,0. 248 for HPA15a/15a, HPA15a/15b, HPA15b/15b, respectively. HPA-3 showed the second greatest heterozygosity with a gene frequency of 0. 339,0. 467, 0. 194 for HPA3a/3a, HPA3a/3b, HPA3b/3b, respectively. HPA genotype frequencies showed a good fit to Hardy-Wein- berg equilibrium. Conclusion Distribution of the allele polymorphism of HLA-A,B and HPA1-17 in Yunnan Han popula- tion is similar to that in southern Han population, and it exhibits its own characteristic. We established the local HLA- 1 and HPA genotyped database of platelet donors in Yunnan.