我国人类免疫缺陷病毒-1主要流行株外膜蛋白基因V3-V4区及其临近区域的特征性氨基酸分析  

Identification of signature amino acids in the env V3-V4 and flanking regions of human immunodeficiency virus type 1 predominant strains in China

作　　者：梁浩[1] 邢辉[1] Jonathan Z.Li 魏民[1] 洪坤学[1] 冯毅[1] 赵全壁[1] 陈建平[1] 全宇[1] 滕涛[1] 邵一鸣 

机构地区：[1]中国疾病预防控制中心艾滋病/性病预防与控制中心 [2]Department of Medicine,University of California,SanFrancisco,USA

出　　处：《中华医学杂志》2005年第13期897-902,共6页National Medical Journal of China

基　　金：国家杰出青年资金资助项目(39925030);国家重点基础研究基金资助项目(G1999054107)

摘　　要：目的鉴定我国HIV-1主要流行毒株亚型的envV3-V4区及其临近区域的特征性氨基酸,并阐明其在追踪传染源和研究疫苗中的作用。方法应用nested-PCR对157份来自我国12个省份的HIV-1毒株env区序列进行扩增,并使用ABI377型测序仪测序,然后应用BLAST、GCG、MEGA和VESPA等生物学软件或程序对env基因V3-V4区及其临近区域序列进行基因型鉴定、系统树分析及特征性氨基酸鉴定。结果157份样本包括54份B′(34·40%)、61份B′/C(38·85%)和42份CRF01-AE(26·75%)毒株。系统树分析结果显示,B′亚型毒株序列均与B·CN·RL42十分接近,B′/C毒株主要与97CN54A和97CNGX6F聚成一簇。而CRF01-AE序列与THCM240和97CNGX2F聚在一起,而且分别聚成明显不同的两个亚组。特征性氨基酸分析发现,我国B′和B′/C毒株分别具有8个保守的特征性氨基酸,而且与代表株的相同位点氨基酸基本一致。而CRF01-AE重组毒株具有11个保守的特征性氨基酸,其中有9个位点与97CNGX2F和TH·CM240不一致。包括这9个特征性氨基酸的样本主要来自除云南省以外的其他省份。结论目前流行于我国的B′和B′/C毒株具有单一的共同传染源,而CRF01-AE毒株可能是通过不同输入源或不同传播途径先后从泰国传入我国的。这将对我国艾滋病防治策略的制定和正在进行的疫苗研究具有重要的指导意义。Objective To identify signature amino acids in the V3-V4 and flanking regions of the env gene from human immunodeficiency virus type 1 (HIV-1) predominant strains in China and to elucidate the role of these signature amino acids on epidemiologic tracking and the development of vaccine.MethodsFragments of the HIV-1 env gene were amplified by nested-PCR from the whole blood of HIV-1 infected individuals from 12 provinces in China.Then, the PCR products were directly sequenced by using ABI 377 DNA SEQUENCER.The sequences covering the env V3-V4 region of the strains were used for the analyses described here.Envelope sequence subtypes were assigned using BLAST (http://www.HIV-Web.lanl.gov). Phylogenetic analyses were performed using GCG and MEGA as well as signature amino acids were identified using VESPA.Results Subtype B′ strains and two recombinants (B′/C and CRF01-AE) were discovered among 157 currently circulating strains in China.The most prevalent subtypes were B′/C (38.85%), followed by B′ (34.40%), and CRF01-AE (26.75%).Phylogenetic tree analysis of env V3-V4 region showed that subtype B′ strains were closely related to B.CN.RL42, while most of B′/C strains clustered with 97CN54A and 97CNGX6F, CRF01-AE strains clustered into two distinct subgroups, which were closely related to THCM240 and 97CNGX2F.Analysis of signature amino acids revealed that eight of positions were identified as conserved signature amino acid sites in the env V3-V4 and flanking regions of the subtype B′ and B′/C strains and almost all signature amino acids were found in their reference strains.Interestingly, eleven signature amino acids were demonstrated in the same regions of the CRF01-AE strains, but nine out of 11 signature amino acid sites were distinct from the same positions of the reference strains 97CNGX2F and TH.CM240.It is noteworthy that these 9 signature amino acids were found in the strains from all of the selected provinces except those from Yunnan province.Conclusion Analysis of the signature amino acids sug

关 键 词：人类免疫缺陷病毒-1 氨基酸分析 特征性 外膜蛋白基因 流行株 HIV-1毒株 nested 基因型鉴定 env基因 流行毒株 方法应用 分析结果 亚型毒株 传播途径 指导意义 疫苗研究 防治策略 传染源 系统树 序列 PCR GCG 生物学 分析及 

分 类 号：R373[医药卫生—病原生物学]