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目的分析哈尔滨市确诊的男男同性恋(MSM)HIV-1感染病例gag基因序列,确定新发感染毒株基因型。方法收集2014年7月至12月确诊且未经过抗病毒治疗的感染者抗凝血,分离外周血单个核细胞,从中提取基因组DNA,采用巢式PCR法扩增gag基因。采用MEGA 5.05软件构建gag基因系统发育树,分析基因型特点。结果共收集49例MSM感染者样本,且均获得gag基因序列。系统发育树显示,49例gag基因主要分布于3个亚型内,其中CRF01_AE亚型39例,占79.6%;B亚型5例,占10.2%;CRF07_BC亚型4例,占8.2%。另外,发现CRF01B重组型1例,占2.0%。在CRF01_AE毒株中,92.3%(36/39)与我国MSM人群参考株成簇,7.7%(3/39)与泰国及我国西南地区异性传播参考株成簇。结论本研究时间段哈尔滨市确诊MSM感染病例以CRF01_AE基因型为主。MSM人群可能成为本地区复杂基因型的重要来源,实时监测该人群新发感染病例的分子流行病学特征,对制定有效地预防和干预措施非常重要。
Objective To analyze the sequence of gag gene in confirmed cases of HIV-1 infection in MSM in Harbin and determine the genotypes of newly infected strains. Methods The anticoagulants were collected from patients with confirmed and non-antiviral therapy from July to December 2014. Peripheral blood mononuclear cells were isolated and genomic DNA was extracted. The gag gene was amplified by nested PCR. The phylogenetic tree of gag gene was constructed by using MEGA 5.05 software to analyze the genotype characteristics. Results A total of 49 cases of MSM infected samples were collected, and gag gene sequence were obtained. Phylogenetic tree showed that 49 cases of gag gene were mainly distributed in 3 subtypes, of which 39 cases were CRF01_AE subtypes, accounting for 79.6%; 5 subtypes B, 10.2% and 4 cases CRF07_BC, accounting for 8.2%. In addition, 1 case of CRF01B recombinant was found, accounting for 2.0%. Among the CRF01_AE strains, 92.3% (36/39) were clustered with reference strains of MSM in China and 7.7% (3/39) were clustered with reference strains of heterosexual transmission in Thailand and Southwest China. Conclusion The genotypes of CRF01_AE were predominant in the confirmed cases of MSM infection in Harbin during the study period. The MSM population may become an important source of complex genotypes in the region. The real-time monitoring of the molecular epidemiology of new infections in this population is important for the development of effective prevention and intervention measures.