上海地区人类免疫缺陷病毒1型感染者原发耐药基因及亚型分析

摘要

本文通过对未经抗病毒治疗患者的人类免疫缺陷病毒1型(HIV-1)毒株进行检测,了解上海地区HIV-1的亚型分布及原发耐药基因变异现状.对118例未经治疗的HIV感染者标本中HIV蛋白酶全长和部分反转录酶基因进行反转录-聚合酶链反应(RT-PCR)扩增,经DNA测序后,进行系统进化树分析和重组分析,以确定HIV-1基因亚型和重组体,并与斯坦福耐药数据库比对,了解耐药突变位点.使用斯坦福REGA HIV亚型分型工具和美国国立生物技术信息中心(NCBI)HIV亚型分析工具分析,获得118例患者的HIV基因序列,基因分型分别为CRF01_AE重组体57例(48.3%)、B亚型36例(30.5%)、CRF07_BC 15例 (12.7%)、CRF08_BC 7例(5.9%)、C亚型2例(1.7%),亚型间或重组体间二重重组体(B/CRF01_AE)1例(0.8%).蛋白酶抑制剂(PI)和反转录酶抑制剂相关的耐药基因突变率达54.2%(64/118),其中2例(1.7%)发生PI耐药,基因突变位点:M46L、Q58E.5例(4.1%)对反转录酶抑制剂产生耐药,其中对核苷类反转录酶抑制剂(NRTI)和非核苷类反转录酶抑制剂(NNRTI)的耐药率分别为3例(2.4%)和5例(4.1%).基因突变位点:NRTI为M41L、D67N、T69I/N/S、K70L、L74V、V75L、V118I、M184V、L210W/F/M/S和T215F;NNRTI为V90I、L100V、K103R/N、V106M/P/I/G、E138G/A、V179E/D/T、Y181C、G190A、H221Y、F227L、K238S和Y318F.结果提示,上海地区HIV毒株以CRF01_AE重组亚型为主,且发现新的重组体,可能出现新重组体流行趋势.PI和反转录酶抑制剂相关的耐药基因突变率较高,且存在高度原发耐药毒株,应加强HIV-1耐药基因变异监测,科学、合理地给予抗病毒治疗.%The present paper aims to better understand the genetic diversity and drug resistance-associated mutations of human immunodeficiency virus type 1 (HIV-1) strains isolated from 118 drug-naive individuals in Shanghai. pol gene from the patients was amplified by nest reverse transcriptase-polymerase chain reaction (RT-PCR) and nucleotide sequencing. REGA HIV-1 Subtyping Tool and National Center for Biotechnology Information (NCBI) HIV Subtyping Tool were used to identify the subtypes of obtained sequences. Drug-resistance-associated mutations in protease and reverse transeriptase regions were analyzed with Stanford University HIV Drug Resistance Database. The results showed that CRF01_AE predominated in Shanghai with 48.3％, followed by subtype B (30.5％), CRF07_BC (12.7％), CRF08_BC (5.9％), and C (1.7％). In addition, an inter-subtype and inter-CRF recombinant B/CRF01_AE (0.8％) strain was detected. Protease inhibitor-associated primary resistance mutations were found in two (1.7％) cases: M46L and Q58E. The mutations conferring primary resistance to nucleoside reverse transcriptase inhibitors (NRTIs) and non-nucleoside reverse transcriptase inhibitors (NNRTIs) were detected in three (2.4％) and five (4.1％) cases, respectively (NRTI: M41L, D67N, T691/N/S, K7OL, L74V, V75L, Vll8I, MI84V, L210W/F/M/S, and T215F;NNRTI: V901, L100V, K103R/N, V106M/P/I/G, E138G/A, V179E/D/T, Y181C, G190A, H221Y, F227L, K238S, and Y318F). This study reveals that CRF01_AE predominates in Shanghai. Antiretroviral drug resistance among untreated HIV-l-infected individuals is 5. 9％. More attention should be paid to the monitoring of HIV-1 drug resistance mutations. Furthermore, drug resistance should be determined in patients with acquired immunedeficiency syndrome (AIDS) before starting antiretroviral therapy.