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目的研究北京市男男性行为者(MSM)艾滋病病毒1型(HIV-1)感染者的毒株亚型、流行现状和变化规律。方法采用单纯随机抽样的方法,抽取北京佑安医院2010-2012年新发现的MSM HIV-1急性期感染者的血样共100人份,采样前均未进行抗病毒治疗。提取外周血单核细胞(PBMC)中的脱氧核糖核酸(DNA),用巢式聚合酶链式反应(PCR)扩增病毒env、gag和pol基因区序列,三区中如有扩增失败,则尝试提取血浆中的核糖核酸(RNA),反转录后进行巢式PCR扩增。PCR产物测序后进行基因亚型分析。计算pol基因区样本间基因离散率,并做系统进化树分析。结果有96份样本的基因序列扩增成功,检出的基因亚型有CRF01_AE、CRF07_BC、B、B′、CRF55_01B、CRF65_cpx和未知亚型,所占比例依次为42.7%、25.0%、15.6%、1.0%、1.0%、3.1%和11.5%。前3种亚型合计占83.3%。各亚型所占比例与相关文献相比有明显变化,表现为CRF07_BC亚型所占比例的快速上升和B亚型所占比例的显著下降。发现了新亚型CRF65_cpx、CRF55_01B和一些未知亚型。结论 2010-2012年,北京市MSM HIV-1急性期感染者的HIV-1毒株亚型分布与相关报道相比已经发生改变,建议对北京地区HIV-1毒株的流行和变异进行密切监测。
Objective To study the subtypes, epidemic status and changes of HIV-1 in MSM in Beijing. Methods A total of 100 blood samples from newly infected MSM HIV-1 acute phase infected patients in Beijing You’an Hospital from 2010 to 2012 were collected by a simple random sampling method. None of the 100 anti-virus samples were taken prior to sampling. DNA was extracted from peripheral blood mononuclear cells (PBMCs), and the env, gag and pol gene sequences were amplified by nested polymerase chain reaction (PCR) Attempts to extract RNA from plasma and reverse transcription followed by nested PCR. PCR products were sequenced for genotyping. Gene polymorphism between pol gene regions was calculated and phylogenetic tree analysis was performed. Results The sequence of 96 samples was successfully amplified. The detected subtypes of CRF01_AE, CRF07_BC, B, B ’, CRF55_01B, CRF65_cpx and unknown subtypes were 42.7%, 25.0%, 15.6% 1.0%, 1.0%, 3.1% and 11.5%. The first three subtypes together accounted for 83.3%. The proportion of each subtype significantly changed compared with the relevant literature, showing a rapid increase in the proportion of CRF07_BC subtype and a significant decrease in the proportion of subtype B. New subtypes of CRF65_cpx, CRF55_01B and some unknown subtypes were found. Conclusion The subtypes of HIV-1 strains in MSM HIV-1 acute phase in Beijing from 2010 to 2012 have been changed compared with the related reports. It is suggested that HIV-1 strains in Beijing should be closely monitored for the prevalence and mutation .