宁夏地区艾滋病患者感染HIV-1病毒pol区基因变异性研究

Moleculo-epidemiological Study on HIV-1 in Ningxia

目的通过巢式聚合酶链式反应检测宁夏地区艾滋病患者感染HIV-1病毒的pol基因变异情况,为艾滋病防治提供参考依据。方法采集HIV-1艾滋病患者全血标本并收集流行病学资料,6h内分离血浆,-70℃保存待用。采用血浆提取HIV-1 RNA,针对HIV-1病毒的pol区PR基因和RT基因设计引物,巢式PCR扩增,扩增产物经凝胶电泳鉴定,片段大小约为1.2kb,鉴定后送公司测序。结果巢式PCR检测样本47份,成功扩增35份样本并获得pol区序列,长1060bp。宁夏地区HIV-1感染者中存在4种基因亚型,23例CRF07_BC,6例CRF08_BC,4例CRF01_AE和2例B亚型,基因离散率为B亚型最大。性传播的18例患者中存在3种重组亚型,CRF07_BC重组亚型为主12例(66.7%);静脉吸毒传播的15例中存在4种重组亚型,CRF07_BC重组亚型为主11例(73.3%),其次为B亚型2例(13.3%),CRFO1_AE重组亚型1例(6.7%)、CRF08_BC重组亚型1例(6.7%),还存在2例性行为兼静脉吸毒的患者,属CRF01_AE重组亚型。结论 HIV-1基因型别趋于多样化,CRF07_BC重组亚型为主要流行毒株,B亚型在宁夏地区流行时间最长;吸毒人群和性活跃人群仍是艾滋病防治的重点人群。

Objective To investigate the characteristics of human immunodeficicncy virus（Hl V） gene sequence,sub-type distribution and origin of Hl V strains,as well as its epidemic status and trends in Ningxia Province,so as to provide scientific evidence for policy making and development of AIDS prevention and control measures. Methods Anticoagulating blood samples were collected from HIV-1 infected individuals and epidemiological data were analyzed. Separation of plasma in 6 hours and -70℃ to save. Design primers according to PR and RT gene of the pol HIV-1 virus,and it was amplified by nested PCR from the RNA extraction and the amplification products is about 1.2 kb,then sequenced. The acquired sequences were compared with the international subtype references. The phylogenetic trees were constructed to determine the subtype of the samples.Results Out of the 47 samples,35 samples were PCR positive and the sequences were acquired. 23 cases were CRF07_BC,6 cases were CRF08_BC,4 cases were CRF01_AE,2 cases were subtype B. The B subtype was the largest genetic-distance. There were three subtypes of 18 patients with sexually transmitted and CRF07_BC was mainly subtype,about 12 cases（66.7%）. There were four subtypes of 15 cases with intravenous drug users and CRF07_BC was mainly subtype,about 11 cases（73.3%）. 2 cases were subtype B（13.3%）,1 case was subtype CRFO1_AE（6.7%）,and 1 case was subtype CRF08_BC（6.7%）. 2 cases with sexually transmitted and intravenous drug use were subtypes CRF01_AE. Conclusion The subtype tends to diversity in Ningxia.CRF07_BC is the dominant strain and shows a high prevalence. Drug users and sex active people still are the key of the HIV/AIDS prevention and control of the crowd.