辽宁省阜新市抗病毒治疗患者HIV-1 DNA耐药情况及亚型分析

DNA drug resistance and subtypes in HIV-1 patients receiving antiretroviral therapy in Fuxin, Liaoning

目的了解阜新市已接受HAART患者HIV耐药株的发生及基因亚型分布情况。方法采集全血标本进行白细胞富集和前病毒DNA提取,巢式PCR扩增HIV-1 pol区基因并测序,使用MEGA以及斯坦福大学HIV耐药数据库等工具对序列信息进行分析。结果在112份全血标本中获得101例有效基因序列,CRF01_AE亚型占84.16%(85/101),CRF07_BC亚型占8.91%(9/101),B亚型占3.96%(4/101),CRF65_cpx亚型占1.98%(2/101),G亚型0.99%(1/101);耐药分析显示27例患者发生耐药位点突变,其中,19例对核苷类逆转录酶抑制剂耐药,主要耐药突变位点为M184I/V,14例对非核苷类逆转录酶抑制剂耐药,主要耐药突变位点为G190S,对两类逆转录酶抑制剂同时耐药的样本有6例,尚未出现蛋白酶抑制剂耐药突变患者;治疗时间以及基线CD4水平的差异对耐药突变具有统计学意义。结论阜新市HIV-1存在5种亚型,CRF01_AE为主要流行亚型;耐药情况的研究为更科学有效地指导艾滋病防治工作提供参考依据,应继续加强耐药监测,掌握基线数据,严防耐药株的流行。

Objective To investigate the prevalence of HIV resistant strains and distribution of subtypes in patients receiving HAART in Fuxin.Methods The whole blood samples were collected for leukocyte enrichment,and its DNA were extracted from each sample.The HIV-1 pol gene was amplified by nested-PCR and the PCR products were subjected to sequencing.The sequences was analyzed by the software MEGA and online tool HIV Drug Resistance Database provided by Stanford University.Results Pol gene fragments were successfully amplified in 101 from 112 samples,of which CRF01_AE accounted for 84.16%(85/101),CRF07_BC 8.91%(9/101),B 3.96%(4/101),CRF65_cpx 1.98%(2/101) and G 0.99%(1/101);27 samples with drug resistant mutations were found,19 of them were resistant to Nucleoside Reverse Transcriptase Inhibitors and the main mutation site was M184 I/V,14 of them were resistant to Non Nucleoside Reverse Transcriptase Inhibitors and the main mutation site was G190 S,6 of them were resistant to both Nucleoside Reverse Transcriptase Inhibitors and Non Nucleoside Reverse Transcriptase Inhibitors,none of them was resistant to Protease Inhibitos;There were significant differences in treatment time and baseline CD4 levels.Conclusion There are five subtypes of HIV-1 in Fuxin City,with CRF01_AE as the major subtype.The research to drug resistance provides reference for guiding AIDS prevention and treatment more scientifically and effectively.It is necessary to strengthen the monitoring of drug resistance and baseline data to strictly prevent the spread of resistant mutant.