AIDS患者高效一线抗逆转录病毒治疗失败后HIV耐药基因型分析  

Genotypic analysis on drug resistance of patients after failure of first-line highly active antiretroviral therapy

作　　者：李彦媚[1] 赵红心[2] 周海卫[2,3] 肖江 张雯[2] 黄英秀[1] 曾辉[2,3] 

机构地区：[1]北京大学地坛医院教学医院,北京市100015 [2]首都医科大学附属北京地坛医院 [3]新发突发传染病研究北京市重点实验室

出　　处：《中华实验和临床感染病杂志（电子版）》2013年第1期68-72,共5页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：北京市朝阳区艾滋病和病毒性肝炎等重大传染病综合防治示范区建设研究(No.2012ZX10004-904);二线抗病毒药物对艾滋病治疗失败患者单核细胞的影响(No.2009-3153)

摘　　要：目的研究一线高效抗逆转录病毒治疗(HAART)失败的艾滋病患者HIV基因型耐药突变情况。方法采集就诊于首都医科大学附属北京地坛医院接受一线HAART治疗失败的共30例艾滋病患者的外周血,分离血浆后,应用巢式PCR方法扩增HIV蛋白酶1～99个氨基酸和逆转录酶前300个氨基酸序列基因,PCR产物经纯化后测序,将获得的序列与美国斯坦福大学耐药数据库比对,得出患者对药物的耐药性解释。结果 30例患者血浆中,扩增成功并得到测序结果28例,扩增成功率为93%(28/30)。其中,27例患者存在耐药突变。在逆转录酶(RT)区检测到48种耐药突变,其中M184V、D67G、K70R、K70K/R、A62V、K219E、K65R、V75I、T215F、D67N是核苷类逆转录酶区发生率>10%的突变,Y181C、G190A、K103N、V179D是非核苷类逆转录酶(NRT)区发生率>10%的突变;在蛋白酶(PR)区检测到A71T、Q58E、A71V、N83D/N4种次要突变。在核苷类逆转录酶抑制剂(NRTI)中,89%的患者出现了对拉米夫定(3TC)和恩曲他滨(FTC)的高、中度耐药,对齐多夫定(AZT)、司他夫定(D4T)、替诺福韦酯(TDF)等也出现不同比例的高、中度耐药;其中TDF和AZT的高、中度耐药率分别是14%和29%。在非核苷类逆转录酶抑制剂(NNRTI)中,50%以上的患者出现对依非韦伦(EFV)、奈韦拉平(NVP)等的高中度耐药;蛋白酶区的次要突变引起了蛋白酶抑制剂(PI)类药物的潜在次要突变,而未出现高、中度耐药。结论患者抗病毒治疗失败主要是发生了对NRTI和NNRTI的耐药,更换二线治疗方案时,TDF可能比AZT更适于和PI联合进行抗病毒治疗。Objective To study the genotypic drug resistance mutations of HIV after the failure of first-line highly active antiretroviral therapy （HAART） in patients with AIDS. Methods Total of 30 HIV- infected patients＇ peripheral blood and separated blood plasma who were treated in Beijing Ditan Hospital, Capital Medical University and failed after first-line HAART were collected, and nested-PCR was taken to amplify the genome sequence of the 1-99 amino acid of HIV protease and the first 300 amino acid of the reverse transcriptase, then PCR products were sequenced after purification, and the acquired nueleotide sequences were compared with resistant database of Stanford University and the interpretation of patients＇ drug resistance was acquired. Results There were 28 cases sequenced successfully among the 30 patients＇ plasma and the amplification rate was 93%. Twenty-seven patients were found drug resistant mutations. Total of 48 drug resistance mutations in the reverse transeriptase （RT） area were detected, wherein the mutation rates of M184, D67G, KTOR, KTOK/R, A62V, K219E, K65R, V75I, T215F and D67N were greater than 10% of the incidence in nucleoside reverse transcriptase （NRT） area. Y181C, G190A, K103N, V179D occured greater than 10% in the non-nucleoside reverse transcriptase （NNRT） area. A71T, Q58E, AT1V, N83D/N were the 4 minor mutations detected in the protease （PR） region. In the nucleoside reverse transcriptase inhibitor （NRTI）, 89% patients were developed to high or intemediate-level of drug resistance to 3TC and FTC, which also appeared in a different proportion to ABC, AZT, IMT, DDI, TDF, and the high or intermediate level of TDF and AZT resistance rates were 14% and 29%, respectively. Among non-nucleoside reverse transcriptase inhibitor （NNRTI）, more than 50% patients had high or intermediate drug resistance to EFV, ETR, NVP, RPV. The minor mutations in PR region induced potenitially low level drug resistance and with none high or intermediate level of protease inhibitors �

关 键 词：高效抗逆转录病毒治疗 耐药 

分 类 号：R512.91[医药卫生—内科学]