广西防城港市HIV-1流行株env基因C2V3区序列特征及亚型分析

Sequence characteristics and subtype analysis of C2V3 region of env gene among HIV-1 strains in Fangchenggang City,Guangxi

目的了解近期防城港市HIV-1流行毒株env基因C2V3区亚型及分布特征,为防城港市防治艾滋病工作提供科学依据。方法于2017年10月～2018年4月在防城港市人民医院收集199例定点随访的接受抗病毒治疗的HIV感染者或艾滋病病人(human immunodeficiency virus/acquired immunodeficiency syndrome,HIV/AIDS)的流行病学调查信息和10 ml外周血。从外周血白细胞中提取前病毒DNA,用巢式聚合酶链反应对获得的HIV-1前病毒DNA env C2V3区基因片段进行扩增,并进行DNA测序,获得的DNA序列信息使用Sequencher 5.1、MEGA 5.0软件进行清理和分析。结果本研究共收集199例HIV/AIDS,其人口学特征主要以年龄18～49岁为主,占53.70%;职业以农民和待业为主,分别占42.59%、45.37%;学历以初中及以下为主,占89.81%;民族以汉族为主,占75.93%;传播途径以异性传播为主,占93.52%。成功扩增HIV-1env C2V3区基因108例,其中CRF01_AE(Circulating Recombinant Forms,CRF)亚型103例,CRF BC亚型3例,B、CRF59_01B亚型各1例。发现8种gp120 V3环顶端四肽,其中以GPGQ为主,占50.00%;其次为GPGR和GPGK,分别占22.22%、12.96%。预测辅助受体为CXCR4(chemo-kine receptor 4) 59例,占54.63%;其次是CCR5辅助受体(chemo-kine receptor 5)30例,占27.78%;CXCR4/CCR5辅助受体19例,占17.59%。结论防城港市主要流行HIV-1 CRF01_AE亚型,其流行毒株亚型存在多态性。防城港市HIV-1型CRF01_AE毒株膜区V3环氨基酸顶端四肽存在较高的多态性,使用CXCR4辅助受体的HIV-1毒株比例高于用CCR5辅助受体的毒株。

Objective To understand the subtypes and distribution characteristics of the C2 V3 region of the HIV-1 strain in the recent city of Fangchenggang,and provide a scientific basis for the prevention and treatment of AIDS in Fangchenggang City.Methods An epidemiological survey of 199 patients with HIV/AIDS who received anti-viral treatment at a fixed-location follow-up was conducted at People’s Hospital of Fangchenggang City from October 2017 to April 2018.The proviral DNA was extracted from peripheral blood leukocytes,and the obtained HIV-1 proviral DNA env C2 V3 region gene fragment was amplified by nested polymerase chain reaction and subjected to DNA sequencing.The obtained DNA sequence information was obtained using Sequencher 5.1 and MEGA 5.0 software for cleanup and analysis.Results A total of 199 HIV/AIDS cases were collected in this study.The demographic characteristics were mainly the age of 18 to 49,accounting for 53.70%.The occupations were mainly farmers and unemployed,accounting for42.59%and 45.37%respectively.The education is mainly junior high school and below,accounting for 89.81%;the ethnic group is mainly Han,accounting for 75.93%;the transmission route is mainly heterosexual transmission,accounting for 93.52%.A total of108 cases were successfully amplified,including 103 cases of CRF01_AE(Circulating Recombinant Forms,CRF)subtype,3 cases of CRF BC subtypes,1 case of B and CRF59_01 B subtypes respectively.During the study,we found 8 kinds of gp120 V3-loop tip tetrapeptides.GPGQ,GPGR and GPGK take up 50.00%,22.22%,12.96%respectively.The predictive co-receptor was CXCR4 in59 cases,accounting for 54.63%.The second is the CC chemo-kine receptor 5 in30 cases,accounting for 27.78%.There were 19 cases of CXCR4/CCR5 coreceptors,accounting for 17.59%.Conclusion CRF01_AE subtype is dominant HIV-1 epidemic strain in Fangchenggang City and its epidemic strain subtype is polymorphic.There is a high polymorphism in the V3 cyclic amino acid tip tetrapeptide of HIV-1 type CRF01_AE strain in Fangchenggang City.The proportion of HIV-1 strain using CXCR4 co-receptor is higher than that of CCR5 co-receptor.