重庆市新确证HIV-1感染者传播性耐药突变研究

Surveillance of HIV-1 transmitted drug resistance mutation among newly diagnosed HIV-1 individuals in Chongqing

目的通过对重庆市新确证人类免疫缺陷病毒1型(HIV-1)感染者耐药基因型的研究,了解我市耐药病毒株的传播水平。方法采集2014年5月至2017年6月我院新确证未治疗的HIV-1感染者的外周血175例,分离血浆后,使用一步法RT-PCR和巢式PCR扩增HIV-1pol区的基因,PCR产物经纯化后测序,将获得的基因序列与美国雅培公司的耐药数据库对比,得出患者对药物的耐药性解释。结果 PCR扩增阳性且测序成功168例,发现16例患者存在耐药突变,传播性耐药突变检出率达9.5%(16/168),其中2014-2017年的耐药比例分别为7.0%、8.0%、9.1%、12.5%,属中度流行。非核苷类反转录酶抑制剂(NNRTIs)、核苷类反转录酶抑制剂(NRTIs)和蛋白酶抑制剂(PIs)耐药突变率分别为9.5%、2.4%和0%。V179D/E(7.1%)和M184V(2.4%)分别是出现最多的NNRTI和NRTI耐药突变位点。168例感染者中,CRF07_BC亚型是主要流行亚型(42.8%),其次为CRF01_AE(38.7%)和CRF08_BC亚型(10.1%)。结论重庆市新确证未治疗HIV-1感染者耐药比例有逐年上升的趋势,应加强监测,预防原发性耐药和耐药毒株的传播。

Objective To investigate the prevalence of HIV-1 drug-resistant strains by exploring the genotypes of the resistance genes in newly diagnosed HIV type1-infected patients in Chongqing city. Methods Plasma specimens of 175 newly diagnosed and antiretroviral treatment-naive HIV-1 infected patients were collected from our center during May 2014 to June 2017. After plasma and serum separation, one-step RT-PCR and nested PCR were taken to amplify the pol gene, then PCR products were sequenced after purification, and the obtained nucleotide sequences were compared with resistance database of the United States Abbott to acquire the interpretation of drug resistance. Results A total of 168 samples were successfully amplified and sequenced. Sixteen patients were found with drug resistant mutations. The total prevalence of transmitted antiretroviral drug resistance （TDR） was 9.5%, and the drug resistance rate was 7.0%, 8.0%, 9.1% and 12.5% respectively in the years of 2014 to 2017, which was classified as moderate. The resistance mutation rates of non-nucleoside reverse transcriptase inhibitors （NNRTIs）, nucleoside reverse transcriptase inhibitors （NRTIs） and protease inhibitors （PIs） were 9.5%, 2.4% and 0%, respectively. V179D/E （7.1%） and M184V （2.4%） were the most commonly emerged NNRTI and NRTI resistance mutations. Among the 168 patients, CRF07_BC was the predominant subtype, accounting for 42.8%, followed by CRF01_AE （38.7%） and CRF07_ BC （10.1%）. Conclusion The rate of TDR is increasing year by year among newly diagnosed, antiretroviral treatment-naive patients in Chongqing city. So it is necessary to do HIV-1 resistance genotyping test for those patients.