[中国本土人群中HIV-1共受体CCR5新突变位点的鉴定及894C缺失变异基因分型]

[Identification of new mutant sites and 894C deletion variant genotyping of HIV-1 coreceptor CCR5 in indigenous Chinese populations].

作者信息

Liu Mingxu, Wang Fusheng, Hong Weiguo, Wang Bo, Jin Lei, Lei Zhouyun, Hou Jing

机构信息

PLA Key Laboratory for Prevention and Therapy of Hepatitis & AIDS, 302 Hospital of PLA, Beijing 100039, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2002 Nov 10;82(21):1468-72.

PMID:12509908

Abstract

OBJECTIVE

To investigate the new single nucleotide polymorphism (SNP) sites of HIV-1 coreceptor CCR5 gene and conduct genotyping of CCR5-894C deletion allele in Chinese populations.

METHODS

The full length fragment of CCR5 gene coding region was amplified by PCR amplification and sequenced in 45 healthy subjects from Han ethnic group. The sequencing data was analyzed by using DNAstar software for identification of new SNP sites. Furthermore, the genotyping of CCR5-894C deletion variant was performed by PCR-RFLP assay in 627 indigenous Chinese individuals including HIV-1 carriers, HIV-1 high risk population of STDs or IDUs as well as normal contrast.

RESULTS

Totally six SNPs and one cytosine base deletion at the 894 nucleotide of CCR5 gene coding region were found in forty five Chinese Han subjects. Four SNPs, i.e. 184A-->G, 503G-->T, 668G-->A, 999G-->T, cause alteration of corresponding amino acids in CCR5 protein and two SNPs are nonsense mutations. The 894C deletion mutation results in a frame shift mutation of CCR5 gene. Among the six SNPs identified, the three sites 184A-->G, 503G-->T, 999G-->T were firstly reported only in Chinese people. The allelic frequencies of mutant 184G, 503T and 999T alleles were 1.1%, 21.1% and 10.0% in Han healthy subjects, respectively. PCR-RFLP assay showed the frequencies of CCR5-894C deletion alleles are 1.11%, 0.53% and 0% for Chinese Han, Tibetan and Mongolian healthy individuals, respectively. There was not significant different of the frequencies of CCR5-894C deletion allele among HIV-1 high risk populations, HIV-1 carries and healthy subjects.

CONCLUSION

The subjects from Chinese Han group has its own distinctive SNP sites in the HIV-1 coreceptor CCR5 gene, three new SNPs (184A-->G, 503G-->T, 999G-->T) were firstly identified. Taking together, the significance and implication of these distinctive SNP sites and the 894C deletion mutation in the pathogenesis of HIV/AIDS disease need to be further studied.

摘要

目的

研究人类免疫缺陷病毒1型（HIV-1）共受体CCR5基因新的单核苷酸多态性（SNP）位点，并对中国人群中CCR5-894C缺失等位基因进行基因分型。

方法

采用聚合酶链反应（PCR）扩增45名汉族健康受试者的CCR5基因编码区全长片段并测序。利用DNAstar软件分析测序数据以鉴定新的SNP位点。此外，采用聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP）方法，对627名中国本土个体（包括HIV-1携带者、性传播疾病或注射吸毒的HIV-1高危人群以及正常对照）进行CCR5-894C缺失变异体的基因分型。

结果

在45名中国汉族受试者中，共发现CCR5基因编码区6个SNP以及894位核苷酸处1个胞嘧啶碱基缺失。其中4个SNP，即184A→G、503G→T、668G→A、999G→T，导致CCR5蛋白相应氨基酸改变，另外2个SNP为无义突变。894C缺失突变导致CCR5基因发生移码突变。在鉴定出的6个SNP中，184A→G、503G→T、999G→T这3个位点为首次仅在中国人群中报道。汉族健康受试者中，突变型184G、503T和999T等位基因的频率分别为1.1%、21.1%和10.0%。PCR-RFLP分析显示，汉族、藏族和蒙古族健康个体中CCR5-894C缺失等位基因的频率分别为1.11%、0.53%和0%。HIV-1高危人群、HIV-1携带者和健康受试者中CCR5-894C缺失等位基因频率差异无统计学意义。