维吾尔族人群HLA-Cw等位基因遗传多态性研究

The genetic polymorphism of Uygur population at HLA-Cw locus by sequence based typing

目的研究新疆维吾尔族人群HLA-Cw等位基因的遗传多态性。方法采用自行建立的HLA-Cw基因测序分型方法,并辅以Ariza商品化测序试剂盒,对100份维吾尔族无关个体血样进行HLA-Cw基因的第2和第3外显子进行序列测定;用ABI PrismTM3100检测测序反应产物,Assign3.5分析软件分析结果。结果(1)经As-sign3.5软件分析,能直接分析出明确的HLA-Cw等位基因型的样本占30%;排除罕见等位基因后,可判定HLA-Cw等位基因型的样本占33%;其余的样本经PCR-SSP高分辨分型试剂盒检测后,可判定出等位基因型;(2)在70份模棱两可的结果中,共有41种NMDP Allele Code,出现120次,其中频率在5%以上的有03BPSK等7种,其频率之和>50%(61/120);(3)检出了27种HLA-Cw等位基因,频率大于10%的2种常见等位基因为:Cw*0602>Cw*0102,介于5%—10%的依次为:Cw*0401>Cw*0303>Cw*0701>Cw*1502,维吾尔族中Cw*02,04,05,06组与Cw*01,03,07,08组的频率分别为0.295和0.485;(4)共检出62种HLA-Cw等位基因型,基因型频率的分布符合Hardy-Weinberg定律。结论本文的结果可为骨髓库和临床无关供/受者样本的HLA-Cw高分辨基因分型提供重要参考;所得到的HLA-Cw位点的等位基因频率可为人类学、遗传学等研究提供基础数据。

Objective To study the genetic polymorphism of Uygnr population at HLA-Cw locus by sequence based typing （ SBT）. Methods A total of 100 unrelated blood samples from Uygnr population were subjected to sequencing exons 2 and 3 of HLA-Cw gene by the SBT established by our laboratory, and 50 samples randomly selected among those 100 samples were sequenced as a parallel control by using the Ariza AlleleSEQR commercial kit. The purified products of sequencing reaction were run eletrophorized on ABI 3100 DNA Sequencer and the designation of HLA-Cw genotype was accomplished by using the Assign3.5 software. Results 1 ） The heterozygous sequence at exons 2 and 3 of HLA-Cw gene for each sample was imported into Assign3.5 software and 30% of the 100 samples could obtain a definite genotype. And 33% of these 100 samples with ambiguous results could be assigned allelic genotype by ruling out the rare alleles according to the NMDP Rare Allele List File; however, only when subjected to further confirmatory testing by PCR-SSP or other molecular method, the remaining 37% of these 100 samples could be defined the final allelic genotype. 2 ） A total of 41 types of NMDP allele codes were observed 120 times in 70 samples with ambiguous result, 7 common NMDP codes with a frequency of more than 5% were found 61 times and the total frequency of these 7 common NMDP allele codes were more than 50% （61/120）. 3 ） Twenty seven HLA-Cw alleles were identified, the common alleles with a frequency of 〉 10% were Cw^＊ 0602 〉 Cw^＊ 0102, four alleles with a frequency ranged from 5% to 10% were listed in order： Cw^＊ 0401 〉 Cw^＊0303 〉 Cw^＊ 0701 〉 Cw^＊ 1502. The gene frequency for Cw^＊02, 04, 05, 06 and Cw^＊01,03, 07, 08 were 0.295 and 0.485, respectively. 4 ） 62 HLA-Cw genotypes at allelic level were determined and the distribution of genotype frequency was in line with Hardy-Weinberg principle. Conclusion Our results can provide useful information for HLA-Cw genotyping by SBT for bone marrow donor and clinical patients. The obtained HLA-Cw allele frequency and its distribution characteristics in Uygnr population could be further applied in anthropology and genetics research work.