HIV-1 gp41基因片段亚型多样性研究

Study on Genetic Diversity of HIV-1 gp41 Gene Segment

〔目的〕应用HIV-1 env gp41基因分型技术进行出入境HIV感染者HIV-1亚型多样性研究。〔方法〕采用套式逆转录PCR方法对env gp41和gp120(C2-V3区)进行基因扩增、测序、亚型对比分析,以及应用威斯康星GCG公司软件进行系统树分析和用美国Los Alamos国家实验室的HIV-Blast软件进行序列的亚型鉴定。〔结果〕对22份样本的gp41和gp120(C2-V3区)的基因亚型进行对比分析,结果表明gp41和gp120(C2-V3区)在亚型上具有一致性。对65例来自中国、非洲、欧洲、美洲和东南亚的出入境HIV-1阳性者进行gp41扩增基因亚型分析,发现有A、B、C、D、F1、G亚型和重组亚型AC、01_AE、02_AG、07_BC、08_BC、06_cpx、18_cpx共计13种亚型,其中重组株占33.8%(22/65)。中国籍出入境感染者包括了A、B、C、D、F1、G亚型和重组亚型01_AE、02_AG、07_BC、08_BC在内的10种亚型,其中重组株占25%(7/28)。〔结论〕用gp41替代gp120(C2-V3区)进行HIV-1亚型分析较为理想。出入境人群中HIV感染者感染的HIV毒株几乎包括了全球流行的所有亚型,多种国内少见的亚型已经通过国际旅行者传入国内。

Objective To document the genetic diversity of HIV-1 in entry-exit individuals of HIV-1 carriers with gp41 genotyping. Methods Reverse strand cDNA was generated from viral RNA and nested PCR was performed. PCR products were sequenced directly through using the primers. Phylogenetic analysis was generated with the procedures of the GCG Wisconsin Package. For subtype determination, the sequences were compared with reference sequences by use of the HIV Blast genotyping tool. Results The subtype designation was the same for both gp41 and gpl20 regions. 65 HIV-1 positive samples, in total were genetically characterized in the env gp41 region, for all of which thirteen subtypes （A, B, C, D, F1, G and circulating recombinant forms （CR10 AC, 01_AE, 02_AG, 07_BC, 08_BC, 06_cpx, 18_cpx） were available. 22 （33.8%） of 65 samples were circulating recombinant forms. There were ten different subtypes （ subtype A,B,C,D,F1 ,G and CRF01_AE,02_AG,07_BC ,08_BC） in the HIV-1 infected individuals from China. of HIV-1 positives in China, 7 （25%） of 28 samples were circulating recombinant forms. Conclusion Subtyping HIV-1 with env gp41 region was an effective method. There exited high genetic diversity in the entry-exit individuals. Some subtypes, which have not been identified, or rarely identified in China, have been introduced by the international travelers.