中国彝族人群CCR5基因单核苷酸多态性及其与HIV感染的关联研究  

Single nucleotide polymorphisms of HIV coreceptor CCR5 gene in Chinese Yi ethnic group and its association with HIV infection

作　　者：马丽英[1] 洪坤学[1] 鲁晓知[2] 秦光明[3] 陈健平[1] 陈康林 阮玉华[1] 邢辉[1] 朱家鸿[2] 邵一鸣 

机构地区：[1]中国疾病预防控制中心性病艾滋病预防控制中心,北京100050 [2]武汉生物制品研究所 [3]四川省疾病预防控制中心 [4]四川凉山彝族自治州皮防站

出　　处：《中华医学杂志》2005年第45期3181-3185,共5页National Medical Journal of China

基　　金：国家十五科技攻关资助项目(2001BA705B02);国家973重点基础研究发展规划资助项目(G1999054107)

摘　　要：目的对中国彝族人群CCR5基因调节区和结构区的单核苷酸多态性(SNP)进行全面检测,并分析SNP与HIV感染的关联。方法通过巢式PCR分别扩增102份彝族正常人和68份HIV感染者样本的CCR5基因调节区和结构区,利用变性高压液相色谱法(DHPLC)筛选存在突变的基因片段,对突变片段进行双向DNA序列测定,然后应用SequenceNavigator软件对CCR5基因的单核苷酸多态性进行分析,并通过SPSSandPPAP软件分析SNPs与HIV感染的相关性。结果在中国彝族人群CCR5基因的编码区中检测到A77G、G316A、T532C、C921T、G668A五种SNP以及第686688位的AGA缺失突变(686Del3),除C921T(Y307Y)为无意义突变外,其他A77G、G316A、T532C、G668A均为有意义突变,它们的氨基酸突变分别为K26R、G106R、C178R、R223Q突变。在这些突变位点中,只有G668A突变的等位基因频率较高,可达0.08,其余均小于0.01,并且在正常人群和HIV感染者两个群体中的等位基因频率没有显著性差异(P>0.05);与编码区所不同的是,在调节区检测到T58934G、G59029A、T59353C、G59402A和C59653T五种SNP,他们的等位基因频率较高,在0.19120.2941范围之间。所检测到的每一个突变在正常人群和HIV感染人群中的分布均差异无统计学意义(P>0.05)。突变位点间的连锁与HIV感染的相关性,提示T59353CG59402A单体型与HIV感染可能存在关联。结论DHPLC是一种快速的高效筛选未知基因突变的高通量方法。SNP与HIV感染的关联性有待于进一步深入研究。Objective To investigate the single nucleotide polymorphism （SNP） of HIV-1 coreceptor CCR5 gene in Chinese Yi ethnic group and the association between these SNPs and HIV/AIDS. Methods Peripheral blood samples of 102 HIV negative persons of Chinese Yi nationality, 87 males amd 15 females, aged 23 （ 12 - 37） , and 68 HIV carriers, 61 meales and 7 females, aged 27 （ 17 - 51 ）. The regulatory and structrual regions of the HIV coreceptor CCR5 gene were amplified from the genomic DNA by nested PCR, each of the two regions was divided into three gene fragments which were overlaped. High throughput DHPLC was used for screening of unknown mutations in each gene fragment. The PCR products showing different peak traces from wild types in DHPLC were sequenced by forward and reverse primers respectively. The sequences were analyzed with the help of Sequence Navigator software to search for SNP loci. Statistical analysis by SPSS and PPAP softwares were made to study the association between these SNPs and HIV infection. Results Five SNPs（A77G, G316A, T532C, C921T, and G668A）and a AGA deletion of the 686-688 nucleotides were discovered in the coding region of this gene in Chinese Yi ethnic group.C921T mutation was a nonsense mutation, and the other SNPs （A77G, G316A, T532C, and G668A） are sense mutation, with the amino acid changes of K26R, G106R, C178R, and R223Q. Only the frequency of R223Q allelie gene was high （0.08） but those of the others were low （ less than 0.01 ）. There was no significant difference in the allele frequaney between the HIV negtive and HIV positive groups （ all P 〉 0.05）. Five SNP loci （T58934G, G59029A, T59353C, G59402A, and C59653T）were found in the regulatory region of CCR5 gene with high allelie frenqueneies of 0. 1912 -0. 2941. Between the HIV negtive and HIV positive groups, there were no differences in the SNP loe （ all P 〉0.05 ）. Statistical analysis of the association between the linkage of mutation loci with HIV infection suggested a significant differ

关 键 词：获得性免疫缺陷综合征 受体 CCR5 彝族 多态性 单核苷酸 

分 类 号：R512.91[医药卫生—内科学]