HIV-1在母婴传播的个体间病毒基因亚型和准种的差异性的实验研究  

Differences of HIV-1 genotypes and quasispecies in the individuals with vertical transmission

作　　者：黄广宇[1] 王宇明[1] 

机构地区：[1]第三军医大学西南医院感染病研究所,重庆400038

出　　处：《大连大学学报》2006年第4期19-23,共5页Journal of Dalian University

摘　　要：[目的]通过对H IV-1在母婴传播的个体间病毒基因亚型和准种的差异性研究揭示H IV-1在母婴传播过程中的变异规律.[方法]提取H IV-1感染母亲及其子女共4例的血浆中总RNA,通过逆转录多聚酶链反应(RT-PCR)获得H IV-1 env基因的C2-V5区域片段,纯化后连接至T载体,转化至Top10大肠埃希菌内增殖,通过蓝白筛选获得阳性克隆,阳性克隆为模板行PCR,产物进行构象敏感性凝胶电泳(CSGE)分析,挑选优势准种和劣势准种测序.[结果]第一对母亲准种复杂性较其子女低,母子之间各准种的核苷酸序列高度同源,均为C亚型;第二对母亲准种复杂性较其子女高,母子之间各准种的核苷酸序列高度同源,母子均为AE亚型.[结论]H IV-1在母婴传播过程中亚型不发生变化,病毒准种在传播过程中可能具有选择性,进入新宿主后因环境变化而被重新筛选,二者可能共同导致优势/劣势准种的变化.此外,母婴之间准种的复杂性与其宿主的免疫状态有关.[ Objective] To disclose the variant hale of HIV-1 vertical transmission by studying the differences of genotypes and quasispecies in the individuals in HIV-1 vertical transmission. [ Methods ] We extracted RNA from the plasma of the individuals of vertical transmission and aquired HIV- 1 env gene C2-V5 DNA segment of env gene by RT-PCR. Purified DNA segment was inserted into T vector and transformed into Topl0 Escherichia coli. We aquired positive clones by blue-white screening and performed PCR using positive clones as models. PCR products were analyses by conformation sensitive gel electrophoresis（CSGE）. The clones of major and minor quasispecies were sequenced. Sequencing results were anylysed. [ Results] HIV-1 quasispecies in the mother of the first group are less complex than her child. The mother and child of the first group are both infected with HIV- 1 of C subtype and their quasispecies nucleotide sequences are highly homologous. HIV- 1 quasispecies in the mother of the second group are more complex than her child. The mother and child of the second group are both infected with HIV- 1 of AE subtype and their quasispecies nucleotide sequences are highly homologous. [ Concludes ] HIV- 1 genentic subtypes are generally not changed in vertical transmission, but HIV-1 quasispecies could be selected in vertical transmission and screened by the new environment of the new host which may both induce the changes of dominative and minor quasispecies. We also find that complexity of HIV- 1 quasispecies is associated with immune status of host.

关 键 词：HIV-1 准种 构象敏感性凝胶电泳 

分 类 号：R373.9[医药卫生—病原生物学]