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

Abstract

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 M184I/V, 14 of them were resistant to Non Nucleoside Reverse Transcriptase Inhibitors and the main mutation site was G190S, 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. 摘要：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。