云南汉族血小板献血者HLA-A、B和HPA基因多态性的分析

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。

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

Objective To investigate the genetic polymorphisms of HLA-A,B alleles and HPA1-17 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) methods were respectively adopted in genotyping HLA-A,B and HPA1-17.Samples of 1 000 unrelated platelet dornors were collected from Kunming,Yunnan province,China.The allele frequencies of HLA-A,B and HPA1-17 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-1b,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-Weinberg equilibrium.Conclusion Distribution of the allele polymorphism of HLA-A,B and HPA1-17 in Yunnan Han population is similar to that in southern Han population,and it exhibits its own characteristic.We established the local HLA-Ⅰand HPA genotyped database of platelet donors in Yunnan.