哈尔滨市新确诊男男同性性接触者HIV-1感染者毒株基因型分析

HIV-1 genotyping analysis among newly diagnosed men who have sex with men in Harbin city of China

目的分析哈尔滨市新确诊男男同性性接触者(men who have sex with men,MSM)HIV-1感染病例的毒株基因型特征。方法收集2012-2015年新确诊且未经过抗病毒治疗的HIV-1感染者抗凝血,分离外周血单核细胞,从中提取基因组DNA,采用巢式PCR法扩增gag和env基因。采用MEGA7.0软件构建gag和env基因系统发育树,分析基因型特点。结果共收集148例MSM HIV-1感染者样本,均获得gag和env基因序列。系统发育树显示,其中CRF01_AE亚型108例,占73.0%;B亚型19例,占12.8%;CRF07_BC 12例,占8.1%;独特重组型9例,占6.1%。在不同年份、不同年龄群体的标本中,CRF01_AE均为优势基因型。在CRF01_AE毒株中,98.1%(106/108)与我国MSM人群参考株成簇,1.9%(2/108)与泰国及我国西南地区异性传播参考株成簇。77.8%(7/9)的独特重组型毒株来自20~40岁群体。2012年开始出现gag和env基因间重组亚型,2014年发现gag基因内重组亚型,2015年出现基因内和基因间均发生重组的复杂亚型,尚未发现env基因内重组毒株。结论哈尔滨市2012年至2015年新确诊男男同性性接触人群HIV-1感染病例中流行毒株基因型较复杂,CRF01_AE亚型毒株一直是本地区的优势毒株,同时,基因间和基因内重组毒株已出现。因此,加强MSM人群新发感染病例的实时监测,并制定有效的预防措施是非常必要的。

Objective To identify the HIV-1 genotypes among newly diagnosed men who have sex with men（MSM）in Harbin city of China. Methods Genomic DNA was extracted from peripheral blood mononuclear cells（PBMCs）of HIV-1-infected MSM who were diagnosed as HIV-1 positive from January of 2012 to December of 2015 and had not received anti-retroviral treatment.The envand gaggene fragments were amplified by nested PCR.Phylogenetic tree was constructed to observe the clustering of envand gagsequences and subsequently analyze the genotyping features of these new infections. Results A total of 148 PBMC samples were collected,and the envand gaggene sequences were successfully obtained from all samples.Multiple genotypes were identified by phylogenetic analysis,including CRF01AE（108/148,73.0%）,subtype B（19/148,12.8%）,CRF07BC（12/148,8.1%）and unique recombinant forms（URFs,9/148,6.1%）.CRF01AE was the dominant genotype in different years and age groups.In the phylogenetic tree,98.1%（106/108）of CRF01AE sequences（either envor gag）clustered with reference genes from Chinese MSM,while1.9%（2/108）gathered with reference genes from Thailand and Southwest regions of China.77.8%（7/9）of URFs were derived from MSM population with the age range from 20 to 40 years old.The intergenic recombination between envand gaggenes emerged in 2012,intragenic recombination among gaggenes appeared from2014,and a complex URF containing these two recombination patterns was identified in a sample collected in2015.No intragenic recombination among env genes was identified. Conclusions Diverse HIV-1 genotypes among newly diagnosed MSM in Harbin city of China were revealed during 2012 and 2015.The CRF01AE is the predominant genotype,and intergenic and intragenic recombinants have emerged among MSM population in this city.