河南省中部地区艾滋病抗病毒治疗失败患者HIV-1耐药基因突变分析

An analysis of HIV-1 mutations conferring drug resistance in non-responders to highly active antiretroviral therapy(HAART)in central Henan Province

目的分析河南省中部地区艾滋病一线抗病毒治疗失败患者HIV-1耐药基因突变情况。方法采集河南省一线抗病毒治疗〉6个月,病毒载量≥1000拷贝/ml的309例艾滋病患者血样标本,提取病毒RNA,通过逆转录及巢式PCR扩增HIV-1pol区基因,测序后提交斯坦福大学HIV耐药数据库进行耐药基因突变分析。结果在扩增的309份pol基因区片段中,RT区主要耐药突变为K103N/S 44.34%（137/309）,其余依次是M184I/V 43.69%（135/309）、T215F/Y 31.07%（96/309）、Y181C29.77%（92/309）、G190A/S 22.01%（68/309）和D67N19.42%（60/309）等。其中患者体内病毒载量为4～5lg拷贝/ml和〉5lg拷贝/ml较3～4lg拷贝/ml时更不易出现M184I/V耐药突变（OR值分别为0.569和0.316,P〈0.05）,K103N/S、M184I/V、M41L和K219Q/E/R/N突变位点在治疗时间〉48个月时发生相应耐药突变比例较治疗时间〈24个月时升高（OR值分别为3.135、2.620、2.413和3.605,P〈0.05）。有79.61%、64.72%和0.97%的患者感染的HIV对非核苷类逆转录酶抑制剂（NNRTIs）、核苷类逆转录酶抑制剂（NRTIs）和蛋白酶抑制剂（PIs）产生耐药。结论艾滋病抗病毒治疗失败者HIV-1基因耐药性突变位点多,比例高,类型较复杂,因此应根据耐药突变情况选用适合的抗病毒治疗方案,以减少耐药毒株的产生。

Objective To analyze HIV-1mutations conferring drug resistance in non-responders to highly active antiretroviral therapy（HAART）in central Henan Province. Methods Serum specimens were collected from 309 patients with AIDS who had a viral load≥ 1000copies/ml and who had been treated with HAART for over 6months.The HIV-l pol gene was amplified using a reverse transcription-polymerase chain reaction（RT-PCR）,and its sequence was then submitted to the Stanford University HIV drug resistance database to analyze the prevalence of mutations producing drug resistance. Results In the pol fragments from the 309 specimens,the most common mutation in the RT region was K103N/S（137/309 specimens,44.34%）,followed by M184I/V（135/309,43.69%）,T215F/Y（96/309,31.07%）,Y181C（92/309,29.77%）,G190A/S（68/309,22.01%）,and D67N（60/309,19.42%）.Univariate logistic regression analysis indicated that the M184I/V mutation was less likely to occur in patients with a viral load of 4-5lg copies/ml（OR=0.569,P〈0.05）or a viral load 〉5lg copies/ml（OR= 0.316,P〈0.05）than in patients with a viral load of 3-4lg copies/ml.The K103N/S mutation（OR=3.135,P〈0.01）,the M184I/V mutation（OR=2.620,P〈0.01）,the M41 L mutation（OR=2.413,P〈0.05）,and the K219Q/E/R/N mutation（OR=3.605,P〈0.01）were more likely to occur after more than 48 months of HAART in comparison to treatment for less than 24 months.Of the patients studied,79.61% were infected with HIV that was resistant to non-nucleotide reverse transcriptase inhibitors（NNRTIs）,64.72%were infected with HIV that was resistant to nucleotide reverse transcriptase inhibitors（NRTIs）,and 0.97% were infected with HIV that was resistant to protease inhibitors（PIs）. Conclusion HIV-1drug resistance was highly prevalent among non-responders to HAART in Henan.HIV-1had a complex patterns of mutations causing drug resistance.More suitable antiretroviral treatment regimens must be selected to decrease the emergence of drug-resistant strains.