浙江省主要地区抗病毒治疗病人HIV毒株耐药基因变异研究

Study on mutation of drug resistant gene in HIV infected patients with antiretroviral therapy in the major regions in Zhejiang Province

目的:调查分析浙江省主要地区(杭州、宁波和温州)的抗病毒治疗病人的耐药基因变异情况,以进一步指导抗病毒药物的选用。方法:选取三地区接受抗病毒治疗的274例患者,对病毒抑制失败(即病毒载量大于1000 IU/ml)的样本进行HIV蛋白酶(PR)全长和部分逆转录酶(RT)基因区进行RT-PCR和巢式PCR扩增并测序。使用斯坦福大学HIVdb程序在线分析耐药相关突变位点,并分析对各药物的敏感性。结果:病毒未抑制率为10.9%(30/274),最终获得29份序列用于耐药基因分析。发现蛋白酶抑制剂(PI)、核苷类逆转录酶抑制剂(NRTI)和非核苷类逆转录酶抑制剂(NNRTI)最主要耐药突变类型分别为M46I、M184V和Y181C/V,三类药物的耐药率分别为0.4%(1/273),7.3%(20/273)和7.7%(21/273),总耐药率为7.7%(21/273)。21个个体发生耐药,其中4.8%(1/21)的个体对PI耐药,95.2%(20/21)对NRTI耐药,所有人均对NNRTI耐药,90.5%(19/21)的个体表现对NRTI和NNRTI的双重耐药。结论:整体而言,浙江省主要地区抗病毒治疗病人中HIV毒株耐药处于较低水平,但耐药突变呈多样化,耐药个体NRTI和NNRTI双重耐药突出,但对PI药物普遍敏感,应结合实际科学合理用药。

Objective：To investigate the mutation of drug resistant gene in HIV infected patients with antiretroviral treatment in the major regions（Hangzhou,Ningbo and Wenzhou） in Zhejiang Province,so as to provide guidance for appropriate selection of ARV drugs.Methods： A total of 294 blood samples from patients with antiretroviral therapy in the three regions were collected.The entire protease gene and part of the reverse transcriptase gene were amplified by RT-PCR and nest-PCR in the samples with viral load larger than 1000 IU/ml,then sequenced the gene fragments.Mutation of drug resistant gene and drug susceptibility was analyzed by the online tool HIVdb Program developed by Stanford University.Results： Rate of virologic failure in patients was 10.9%（30/274）.29 gene sequences that acquired were used for genotypic resistance analysis.The primary mutation types of drug resistance were M46I,M184V and Y181C/V for PI,NRTI and NNRTI,respectively.Drug resistance rates of PI,NRTI and NNRTI were 0.4%（1/273）,7.3%（20/273） and 7.7%（21/273）,respectively,with the total was 7.7%（21/273）.21 patients were indicated drug resistance,in which 4.8%（1/21） exhibited PI resistance,95.2%（20/21） NRTI resistance,100% NNRTI resistance and 90.5%（19/21）dual NRTI and NRTI resistance.Conclusion： The presence of drug resistant gene in HIV strains was at low level as a whole,but mutation diversity was found in drug resistant gene and the problem of dual NRTI and NNRTI resistance was serious,however general PI susceptibility was found,which calling for the scientific and appropriate selection of ARV drugs.