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目的了解哈尔滨市男男同性恋者(MSM)感染HIV-1流行株的亚型分布状况及基因变异情况。方法采集MSM人群中HIV感染者血液标本,提取病毒RNA并应用巢式聚合酶链反应(nested-PCR)对HIV病毒gag区基因进行扩增,并对扩增产物测序。Mega 5.1软件进行系统进化树分析及基因离散距离的计算,Bio Edit 7.0.0软件将相应的基因序列翻译成氨基酸并分析其变异情况。结果成功获得61条gag基因序列,亚型分析结果显示,CRF01_AE 49例(80.32%),CRF07-BC 5例(8.19%),B亚型7例(11.47%)。其中B亚型基因离散距离最大(0.078±0.005),CRF01_AE最小(0.051±0.004)。CRF01_AE在MSM人群中形成两个独立的进化簇。各亚型的MHR序列变异对比结果显示,B亚型的中MHR序列中第7位缬氨酸(V)突变为异亮氨酸(I)的比例最大。结论哈尔滨市MSM人群中主要存在CRF01-AE、CRF07-BC和B三种亚型,以CRF01-AE为主要流行亚型;其中B亚型基因变异程度最大。对MSM人群HIV-1流行及变异情况的监测有助于对MSM人群开展及时有效的治疗。
Objective To investigate the subtypes distribution and genetic variation of HIV-1 strains in MSM in Harbin. Methods Blood samples from HIV-infected individuals in MSM population were collected, viral RNA was extracted and nested-PCR was used to amplify the gag region of HIV virus. The amplified products were sequenced. Mega 5.1 software for phylogenetic tree analysis and genetic distance calculation, Bio Edit 7.0.0 software will translate the corresponding gene sequence into amino acids and analyze their variation. Results Sixty-one gag gene sequences were successfully obtained. The subtype analysis showed that CRF01_AE was found in 49 cases (80.32%), CRF07-BC in 5 cases (8.19%) and subtype B in 7 cases (11.47%). Among them, the distance of B subtypes was the largest (0.078 ± 0.005) and the smallest was CRF01_AE (0.051 ± 0.004). CRF01_AE forms two independent evolutionary clusters in the MSM population. Comparison of the MHR sequence variations of each subtype showed that the ratio of the valine (V) to the isoleucine (I) at position 7 in the MHR sequence of subtype B was the largest. Conclusion There are three subtypes of CRF01-AE, CRF07-BC and B in MSM population of Harbin, with CRF01-AE as the main epidemic subtype. Among them, the mutation degree of subtype B is the largest. Monitoring the prevalence and variability of HIV-1 in MSM populations can facilitate timely and effective treatment of MSM populations.