四川彝族人群HIV-1辅助受体CCR5△32和CCR2-64I基因多态性分析

Genetic polymorphism of human immunodeficiency virus coreceptor CCR5A32 and CCR2-64I alleles in Chinese Yi Ethnic group in Sichuan

目的 了解中国四川彝族人群艾滋病病毒-1(HIV-1)辅助受体CCR5△32和CCR2-64I基因多态性特点。方法 提取119份彝族正常人和88份HIV-1感染人群外周血基因组DNA。用聚合酶链反应(PCR)方法检测CCR5△32突变,阳性产物经克隆、测序进一步证实;用PCR-限制性片段长度多态性技术检测CCR2-64I突变,并测序验证。结果 119份正常人样本中,CCR5 wt／△32等位基因突变杂合子2例(1 68％),未检测到CCR5△32／△32突变纯合子,CCR5△32等位基因频率为0.0084;CCR2-64I突变杂合子26例(21.85％),突变纯合子2例(1.68％),等位基因频率为0.1261。88份HIV-1感染者样本中,未检测到CCR5△32突变;CCR2-64I突变杂合子12例(13.64％),突变纯合子7例(7.95％),等位基因频率为0 1327。统计分析表明,上述等位基因多态性在该群体中均呈Hardy-Weinberg平衡分布;两种等位基因的突变频率在正常人和感染人群中的差异均无统计学意义。结论研究获得了中国四川彝族人群CCR5△32、CCR2-64I等位基因多态性资料,结果有助于综合评估中国人群对HIV-1感染的遗传易感性,同时为深入研究HIV-1抗性基因在中国不同民族的HIV感染及发病机制中的作用奠定基础。

Objective To explore genetic polymorphisms CCR5 of HIV coreceptor and CCR2 in Chinese Yi Ethnic group in Sichuan. Methods Genomic DNA samples were obtained from 119 healthy individuals and 88 HIV-1 infected individuals of Chinese Yi Ethnic group in Sichuan. Polymerase chain reaction (PCR) , cloning and gene sequencing techniques were employed to identify the genotype of CCR5A32; PCR-restriction fragment length polymorphism(RFLP) and gene sequencing were employed to identify the CCR2-64I alleles. Results At CCR5 locus, 2 heterozygotes (CCR5-wt/A32) and none homozygote (CCR5-A32/A32) were observed m 119 healthy individuals, allelic frequency of CCR5-A32 was 0.84% ; No mutant was found m 88 HIV-1 infected individuals. At CCR2 locus, 26 heterozygotes (CCR2-64V/641) and two homozygotes (CCR2-64I/64I) were observed in healthy individuals but the allelic frequency CCR2-64I was 12.61%. Among infected individuals, 12 heterozygotes(CCR2-64V/64I) and 7 homozygotes (CCR2-64I/64I) were observed and the allelic frequency CCR2-64I was 13.27%. Statistical analysis revealed that the differences of both loci between healthy and infected individuals were insignificant. Both loci were consistent with the Hardy-Weinberg equilibrium in the two different groups. Conclusion The polymorphism of CCR5A32 and CCR2-64I alleles from Chinese Yi Ethnic group was detected which was of Slgnificance for the evaluation of genetic resistance to HIV-1 infection in Chinese population.