献血者血小板1-16抗原基因遗传多态性分析及已知HPA型供者数据库的建立

Analysis of Genetic Polymorphism in Randomized Donor's HPA 1-16 Antigens and Establishment of Typed Platelet Donor Data Bank

本研究的目的是分析人类血小板抗原(humanplateletantigen,HPA)基因多态性,根据分布频率来判断HPA抗原不配合比率以及抗体产生的机会,确定有临床意义的血小板抗原系统,并建立邯郸地区血小板基因频率数据库和供者库。采用SSPPCR方法对邯郸地区148名随机献血者进行HPA116抗原32个等位基因的检测分析,并与不同人群的分布频率进行比较。结果表明:每个样本均检测到HPA1a、2a、4a14a、16a基因;HPA4a、7a14a、16a呈现单态性,未检测出相应的等位基因HPAb;对于HPA1、2、5、6主要以a/a纯合子为多,a/a基因型频率分别是0.9595、0.8108、0.9865、0.9797,没有b/b纯合子出现。在HPA116中,具有最高杂合度的是HPA15,基因型HPA15a/15a、HPA15a/15b、HPA15b/15b频率分别是0.2230、0.5270、0.2500;HPA3在其次,基因型HPA3a/3a、HPA3a/3b、HPA3b/3b频率分别是0.3851、0.5135、0.1014。经χ2检验,结果符合HardyWeinberg遗传平衡定律。邯郸地区随机献血者HPA15系统基因频率与石家庄地区相似(P>0.05);与我国台湾人群进行HPA113、HPA15的比较,HPA1、2、6具有明显的不同(P<0.05),其它相似(P>0.05);与韩国人群进行HPA18的比较,除HPA3具有明显不同外(P<0.05),其余均相似(P>0.05);与美国黑人进行HPA15的比较,HPA1、2、5具有明显的差异(P<0.05);与英国人进行HPA111的比较,HPA1、5具有明显的不同(P<0.05)。结论:北方地区中国人群HPA2、3、5、15系统具有多态性,且HPA抗原分布不配合比率较高,这必然造成免疫暴露的机会增加,提示在临床上可能具有重要的免疫学意义。同时,在此次研究数据的基础上建立了邯郸地区血小板基因频率数据库和血小板已知型供者库。

To study the genetic polymorphism of HPA 1-16 platelet antigen alleles among unrelated volunteer donors and establish a typed platelet donor panel in Handan, typing was perfomed by polymerase chain reaction using sequencespecific primers （SSP-PCR） ; 148 random unrelated blood donors in Handan were genotyped for each of the HPA 1-16 antigen. The gene frequencies were analyzed and the genetype frequencies were determined by direct counting, and these data were compared with HPA distribution among various population by the chi-square test. The results indicated that HPA-1a,2a,4a-14a, 16a genes were found among the 16 HPAs in every sample tested. Monomorphic HPA-4a,7a-14a, 16a were found in the samples. For HPA-1,2,5 and 6, a/a homozygosity was predominant with frequencies of 0.9595, 0. 8108 ,0. 9865 ,0. 9797 ,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. 2230,0. 5270,0.2500 for HPA15a/15a, HPA15a/15b, HPA15b/15b, respectctively. HPA-3 showed the second greatest heterozygosity with a gene frequency of 0. 3851,0. 5135,0. 1014 for HPA3a/3a, HPA3a/3b, HPA3b/3b, respectively. HPA genotype frequencies showed a good fit to Hardy-Weinberg equilibrium. HPA1-5 gene frequencies for Chinese people in Handan were consistent with those of Chinese people in Shijiazhuang（ P 〉 0.05 ）. Among the HPA 1-13, -15, the frequencies of HPA-1, -2, -6 for Chinese people in Handan differed appreciably from those for Chinese people in Taiwan （ P 〈 0.05 ）, others were similar to those of Chinese people in Taiwan. Among the HPA 1-8, a similarity was noted between Chinese people in Handan and Koreans（ P 〉 0.05 ）, except for HPA-3. Frequencies of HPA-1,-2,-5 significantly were differed from those in African Americans, as compared with HPA 1 -5 （P 〈 0.05 ）. Comparison of gene frequencies from HPA-1 and -5 showed significant differences between Chinese people in Handan and people in UK（ P 〈 0.05 ）. It is concluded that HPA-2,-3,-5,-15 of people in Western region of China have polymorphism, incompatible frequency of HPA antigen distribution is higher, which ineviatably results in the increase of immunologic exposure, therefore attention must be paid to the importance of HPA-2,-3 ,-5 ,-15 in clinical disorders . This study for the first time completely analyses HPA1-16 ene frequencis in China,and provides data for establishing a typed platelet donor panel in Handan, China.