河南省HIV-1新近感染者耐药基因研究

Genotypic drug resistance among recently HIV-1 infected bersons in Henan

目的了解河南省艾滋病病毒Ⅰ型(HIV-1)新近感染者的耐药情况。方法2006年8月-2007年6月,在河南省艾滋病自愿咨询(VCT)检测点发现的未进行抗病毒治疗的HIV-1感染者,以酶联免疫吸附试验初筛、蛋白印迹试验确认HIV-1感染,BED-CEIA方法检测新近感染。检出的新近感染样品进行基因型耐药检测,提取血浆中RNA,套式聚合酶链反应(Nested-PCR)扩增HIV-1pol基因区,PCR产物双脱氧法测序,所得序列与LosAlamos HIV标准株序列比对,构建系统进化树分析亚型;利用Stanford HIVdb Drug Resistance Database分析耐药相关突变(DRM)和耐药情况。结果共检出HIV-1新近感染39例,扩增测序有34例新近感染样品分析成功。亚型分析结果为B′亚型32例(94.1%),CRF01_AE重组亚型1例(2.9%),C亚型1例(2.9%)。未发现蛋白酶抑制剂(PI)主要DRM,检测到10例(29.4%)存在PI次要DRM;未发现核苷类逆转录酶抑制剂(NRTI)的DRM;3例(8.8%)存在非核苷类逆转录酶抑制剂(NNRTI)的DRM。耐药分析显示,有2例(5.9%)对NNRTI类药物耐药。结论目前河南省HIV-1新近感染人群中耐药状况处于中等水平,有必要加强HIV-1的耐药监测。

Objective To understand human immunodeficiency virus type 1 （HIV-1） primary resistance profile among recently infected individuals in Henan province. Methods Treatment-naive persons were recruited from August 2006 to June 2007 at local VCT sites. HIV-1 infections were screened by ELISA and confirmed by WB. The recent infection was confirmed by BED-CEIA. The HIV-1 pol gene was amplified by nested-PCR from plasma RNA. The PCR products were sequenced using dideoxynucleoside arrays. Phylogenetic analysis was performed u- sing a set of reference sequences available at the Los Alamos Database to determine subtypes. Edited sequences were analyzed with Stanford HIVdb Drug Resistance Database and drug resistance mutations（DRM） were determined to particular anti-HIV drugs. Results Thirty-nine cases were confirmed recently infected by BED-CEIA. Thirty-four recent samples were successfully analyzed. The circulating strains were B （94.1%）,CRF01_AE （2.9%） and C （2.9%）. No protease inhibitors （PI） major resistance mutation was detected. Ten samples （29.4%） displayed PI minor resistance mutations. No nucleoside reverse transcriptase inhibitors （NRTI） mutation was detected. Three samples （8.8 %） presented DRMs to non-nucleoside reverse transcriptase inhibitors （NNRTI）. The results of the a- nalysis showed that there were 2 samples （5.9 %） which inferred drug resistance to NNRTI. Conclusion The prev- alence of primary drug resistances is at moderate level among recently HIV-1 infected persons in Henan province. Drug resistance surveillance among HIV-1 infected persons should be strengthened.