东莞市HIV-1主要毒株基因分型及其对耐药影响的研究

Genotyping of major HIV-1 strains and its impact on drug resistance in Dongguan

目的了解东莞市人类免疫缺陷病毒Ⅰ型(HIV-1)感染者流行毒株基因亚型的分布特征及耐药情况。方法收集2015年10月至2016年2月东莞市疾病预防控制中心未经抗病毒治疗的50例HIV感染者血浆,提取病毒RNA,采用实时荧光定量逆转录聚合酶链反应(RT-PCR)法扩增pol基因序列并测序,应用BioEdit、Genotyping和MEGA等生物学软件对pol基因进行HIV-1基因亚型分析,并将获得的核酸序列提至美国斯坦福大学HIV耐药数据库(http://hivdb.stanford.edu)进行耐药位点分析。结果 50例样本最终成功扩增得到35份样本的pol基因序列,在这35例研究对象中,88.6%(31/35)为男性,65.7%(23/35)为经性接触感染。35条pol基因序列中,CRF01AE亚型占37.1%(13/35),CRF07BC亚型占37.1%(13/35),B亚型占14.3%(5/35),CRF5501B亚型占11.4%(4/35)。在35条基因序列中检测到11条(占31.4%)含有耐药相关突变,以CRF01AE和CRF5501B重组亚型为主,原发性耐药率为11.4%(4/35)。结论东莞市HIV-1感染者存在着多种基因亚型,以CRF01AE和CRF07BC重组亚型为主,基因序列发生原发耐药性,应引起重视。

Objective To explore the subtype distribution characteristics and drug resistance of human immunodeficiency virus type 1(HIV-1)among HIV-1-infected individuals in Dongguan. Methods Plasma samples of 50 HIV infected individuals without antiviral treatment were collected from October 2015 to February 2016 in Dongguan Municipal Center for Disease Control and Prevention. The viral RNA of HIV-1 was extracted,the pol gene was amplified by nest RTPCR and the PCR products were sequencd. The obtained nucleic acid sequences were submitted to the Stanford University HIV Drug Resistance Database(http://hivdb.stanford.edu)for drug resistance analysis. Results Among the 50 plasma samples,the pol gene sequences of 35 samples were successfully amplified. Of the 35 subjects,88.6%(31/35)were male and 65.7%(23/35)were infected through the sexual transmission. Among the 35 pol gene sequences,37.1%(13/35)were CRF01AE,37.1%(13/35)were CRF07BC,14.3%(5/35)were subtype B,and 11.4%(4/35)were CRF5501 B.Mutations related to drug resistance were found in 11(31.4%)of 35 pol gene sequences. Major mutation subtypes of HIV-1 were CRF01AE and CRF5501 B. The rate of primary drug resistance was 11.4%(4/35). Conclusion The genetic subtypes of HIV-1 were diverse and the main recombinant subtypes were CRF01AE and CRF07BC in Dongguan City. More attention should be paid to the primary drug resistance mutations in gene sequence.