云南省昭通市50岁及以上HIV感染者/AIDS患者抗病毒治疗失败病例耐药和分子网络分析

Drug resistance and molecular network analysis of antiretroviral therapy failure in HIV/AIDS patients aged 50 years and above in Zhaotong City,Yunnan Province

目的了解云南省昭通市2020—2022年50岁及以上抗病毒治疗失败[血浆病毒载量(viral load,VL)≥1000拷贝/mL]人类免疫缺陷病毒感染者/艾滋病患者(human immunodeficiency virus/acquired immunodeficiency syndrome,HIV/AIDS)的耐药及分子网络特征。方法通过云南省抗病毒治疗数据库收集2020—2022年昭通市50岁及以上抗病毒治疗失败的HIV/AIDS患者人口学资料,采集患者血浆,通过逆转录聚合酶链反应扩增HIV pol基因蛋白酶和逆转录酶区,采用ContigExpress软件拼接并进行序列测定,运用美国斯坦福大学HIVdb数据库进行耐药突变分析,利用HyPhy 2.2.4以TN93模型计算序列间的两两基因距离,以1.7%作为阈值构建HIV分子网络,使用Cytoscape 3.7.0将网络可视化。结果共纳入HIV/AIDS病例480例,血浆样本扩增阳性417例,耐药196例,总耐药率为47.0%;非核苷类反转录酶抑制剂(non-nucleoside reverse transcriptase inhibitors,NNRTIs)的耐药率最高为45.6%(190/417),反转录酶抑制剂(nucleoside reverse transcriptase inhibitors,NRTIs)耐药率为14.4%(60/417),蛋白酶抑制剂(protease inhibitors,PIs)的耐药率较低,为0.2%(1/417)。NNRTIs耐药突变位点中,以K103N/E/S突变位点为主,突变率为33.3%(139/417);NRTIs耐药突变位点中,以M184V/I突变位点为主,突变率为13.9%(58/417);PIs耐药位点突变率相对较低,突变率为0.2%(1/417)。通过构建分子网络,88例患者进入分子网络,入网率为21.1%,参与构成21个传播簇,其中CRF01_AE构成了一个包含37例的大簇。结论相较于云南省其他地区研究,本研究昭通市抗病毒治疗失败的老年人群耐药率有所下降,URFs成为主要亚型,提示昭通市老年人传播关系逐渐复杂化,应加强监测检测;通过构建分子网络,提示CRF01_AE、已婚或有配偶男性入网率最高,应加强重点人群干预,多元化开展老年人艾滋病防治工作。

Objective To understand the characteristics of drug resistance and molecular transmission network of HIV/AIDS patients aged 50 years and above in Zhaotong City,Yunnan Province,from 2020 to 2022,who have failed antiretroviral therapy(plasma viral load≥1000 copies/mL).Methods The demographic data of HIV/AIDS patients aged 50 years and above with failed antiviral therapy in Zhaotong City from 2020 to 2022 were collected through the antiviral therapy database of Yunnan Province.The plasma of the patients was collected,and the HIV pol gene protease and reverse transcriptase regions were amplified by reverse transcriptase chain reaction,the fragments were spliced using ContigExpress,the sequences were determined,and drug resistance mutations were analyzed using the HIVdb database of Stanford University.Pairwise genetic distances between sequences were calculated using the HyPhy 2.2.4 software with the TN93 model,and a threshold of 1.7%was used to construct the HIV molecular network.Cytoscape 3.7.0 was used to visualize the network.Results A total of 480 HIV/AIDS cases were included,with 417 positive amplifications of plasma samples.Among these,196 cases were drugresistant,resulting in a total drug resistance rate of 47.0%.The highest resistance rate was observed in NNRTIs at 45.6%(190/417),followed by NRTIs at 14.4%(60/417),and PIs at a lower rate of 0.2%(1/417).In NNRTIs resistance mutation sites,K103N/E/S mutation sites were dominant,with a mutation rate of 33.3%(139/417).Among the NRTIs resistance mutation sites,M184V/I mutation site was the main one,with a mutation rate of 13.9%(58/417).The mutation rate of PIs resistance sites was relatively lower,at 0.2%(1/417).Through the construction of the molecular transmission network,88 patients entered the molecular transmission network,with an access rate of 21.1%,forming 21 transmission clusters,among which CRF01_AE formed a large cluster including 37 individuals.Conclusions Compared with studies in other regions of Yunnan Province,the drug resistance rate among the elderly population with antiretroviral therapy failure in this study in Zhaotong City has decreased,and URFs have become the main subtype,suggesting that the transmission relationships among the elderly in Zhaotong City are gradually complicated.Through the construction of a molecular network,it is suggested that CRF01_AE and married or partnered males have the highest network access rate.Interventions should be strengthened among key populations,and diversified efforts in HIV/AIDS prevention and treatment for the elderly should be developed.