抗HIV治疗失败后HIV1耐药基因的变异

Drug resistance variation in HIV1 after the failure of highly active antiretroviral therapy

摘要:

目的研究抗人免疫缺陷病毒（HIV)治疗失败后，HIV1遗传基因变异情况，并分析其耐药性。方法选取抗HIV治疗失败的57例患者(其中采用D4T/3TC/NVP治疗方案者45例，3TC/AZT/NVP方案者7例，3TC/TDF/克立芝方案者5例），从其血浆中提取病毒RNA，运用逆转录-聚合酶链反应（RTPCR）和套式PCR扩增HIV1 pol区基因片段,并对扩增的目的片段进行测序,将测序结果提交Web站点（http://HIVdb.stanford.edu），分析耐药变异情况。结果3种治疗方案治疗失败者均出现耐药。57例患者，7例发生针对蛋白酶抑制剂的位点变异，其中1例（1.75%）M46IM蛋白酶抑制剂的位点变异，导致了抗ATV/r、FPV/r、IDV/r、LPV/r的潜在低度耐药和抗NFV的高度耐药；32例（56.14%）患者体内HIV1发生了逆转录酶基因变异，且均对不同逆转录酶抑制剂产生了耐受性，其中14例（43.75%，14/32）患者对斯坦福数据库中的11种药物全都耐药，且11例（78.57%）治疗方案为D4T/3TC/NVP。TDF、克立芝等二线药物的治疗方案突变位点少于一线药物D4T、3TC、NVP、AZT。结论耐药变异是导致抗HIV治疗失败的主要原因。在开展抗病毒治疗过程中，应适时进行耐药性检测，以获得较好疗效。

Abstract:

ObjectiveTo study genetic variation and drugresistance of HIV1 after the failure of highly active antiretroviral therapy(HAART) in HIV1/AIDS patients.MethodsFiftyseven patients with HAART failure, including 45 with D4T/3TC/NVP regime，7 with 3TC/AZT/NVP，and 5 with 3TC/TDF/ LPV/r, were enrolled in the study, viral RNA was extracted from plasma, partial HIV1pol gene was amplified by reverse transcription polymerase chain reaction(RTPCR)and nested PCR,and the amplified fragment was sequenced,the phylogenesis of sequences were analyzed by landing the websites http://HIV1db.stanford.edu., and drug resistance variation was analyzed.ResultsPatients with the failure of 3 types of therapeutic regimes all developed drug resistance. Among 57 patients, drugresistant variation in protease inhibitors(PIs) was identified in 7 patients, 1 of whom （1.75%）produced variation in M46IM which resulted in lowlevel resistance to ATV/r,FPV/r,IDV/r,LPV/r and highlevel resistance to NFV; Drugresistant variation in reverse transcriptase inhibitors (RTs) was identified in 32 patients（56.14%）,and 14 （43.75%,14/32）of whom had resistance to 11 kinds of inhibitors(//HIV1db.stanford.edu)，11 (78.57%,11/14) cases were with the regime of D4T/3TC/NVP. Mutation site in patients with secondline drugs(TDF,LPV/r)were less than firstline drugs(D4T,3TC,NVP,AZT). ConclusionDrug resistance variation is the major cause of failure of HIV1 therapy, drug resistance should be monitored during the course of antiHIV therapy.