人类免疫缺陷病毒1型在垂直传播的个体间病毒基因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全军感染病研究所,重庆400038

出　　处：《第三军医大学学报》2005年第17期1785-1788,共4页Journal of Third Military Medical University

摘　　要：目的通过对人类免疫缺陷病毒1型(hum an immunodefic iency virus-1,H IV-1)在垂直传播的个体间病毒基因型和准种的差异性研究揭示H IV-1在垂直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包膜蛋白(env)基因的C2-V5区域片段,纯化后装入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(conform ation sensitive gel electrophoresis,CSGE)分析,挑选优势准种和劣势准种测序。结果第1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2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在垂直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 rule of HIV-1 vertical transmission by studying the differences of genotypes and quasispecies in the individuals with HIV-1 vertical transmission. Methods RNA was extracted from the plasma of the individuals of vertical transmission and C2-V5 DNA segment of HIV-1 env gene was acquired by RT-PCR. Purified DNA segment was inserted into T vector and transformed into Top10 Escherichia coli. Positive clones were acquired by blue-white screening and used as models in PCR. PCR products were analyzed by conformation sensitive gel electrophoresis （CSGE）. The clones of major and minor quasispecies were sequenced. Results HIV-1 quasispecies in the mother of the first group were less complex than her child, and their quasispecies nucleotide sequences were highly homologous and of HIV-1 of C subtype. HIV-1 quasispecies in the mother of the second group were more complex than her child, and their quasispecies nucleotide sequences were highly homologous and of HIV-1 of AE subtype. Conclusion HIV-1 genetic 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[医药卫生—病原生物学] R394.2[医药卫生—基础医学]