Objective: To survey the prevalence of drug resistant HIV-1 in Shandong province in 2013-2015. Methods: WHO truncated sequential sampling technique was adopted by using 77 and 53 samples of newly diagnosed as HIV-1 positive and aged 16-25 years in Shandong province in 2013 and 2015. RNA was prepared and HIV-1 pol region was amplified by RT-PCR and nested PCR. Pol genetic mutation associated with drug resistance was analyzed. Results: The success rates for sequence acquisition of the survey were 100% (77/77) and 94% (50/53) in 2013 and 2015, and the main subtype was CRF01_AE. A total of 2 surveillance drug-resistance mutation(SDRMs) and 3 SDRMs were found by analyzing the 47 sequences each year, sampled in 2013 and 2015, indicating that the prevalence of drug resistant HIV-1 stains was low in 2013, and moderate in 2015. A total of 5 individuals with drug resistant HIV-1 stains found in this study were mainly infected by homosexual transmission (3 cases), and the other two samples were different: one was infected by heterosexual transmission, the other was infected by IDU. The subtype was CRF01_AE (2 cases) , CRF07_BC (2 cases) and B (1 case) . SDRMs for protease inhibitor (PIs), nucleotide HIV-reverse transcriptase inhibitor (NRTIs) and non-NRTI (NNRTIs) were all found in the individuals with drug resistant HIV-1 stains. Conclusion: CRF01_AE were the main HIV-1 subtypes of recently reported HIV-infected individuals in Shandong province, and the HIV-1 drug resistant strains transmission was catalogued as at low and moderate prevalence level in 2013 and 2015.
目的： 调查山东省2013—2015年新发现未治疗HIV-1感染者耐药毒株传播警戒线。 方法： 按照《全国艾滋病病毒耐药性监测指南》中耐药警戒线调查的要求，以2013和2015年新报告的年龄为16~25岁的HIV-1感染者为调查对象，抽取4~8 ml静脉血，用EDTA抗凝，立即混匀，颠倒8~10次，24 h内取0.5 ml全血标本进行CD(4+)T淋巴细胞计数，并提取HIV-1病毒RNA。采用RT-PCR和巢氏PCR方法扩增HIV-1的pol区蛋白酶(PR)和逆转录酶(RT)基因区，PCR扩增产物经电泳并纯化后进行测序。根据WHO的HIV-1耐药警戒线调查方案，采用二项顺序抽样法进行抽样，最大样本量随着抽样和检测的进展而确定，按年度分别将获得有效序列的样本按确证时间先后进行排序，并结合"HIV-1耐药警戒线监测抽样表"进行耐药株传播水平分析。 结果： 2013和2015年调查人数分别为77和53例，调查对象的序列成功获得率分别为100%（77/77）、94%（50/53），亚型均以CRF01_AE为主，分别为53.2%（41/77）、68.0%（34/50）。2次调查分别抽取了47个样本序列，其中发现2份和3份样本存在监测性耐药突变，对应的HIV-1耐药株传播水平分别为低流行（4.3%）和中流行（6.4%）。5例耐药个体的感染方式主要为同性传播（3例）、异性传播（1例）和静脉吸毒传播（1例），亚型分别为CRF01_AE（2例）、CRF07_BC（2例）和B（1例）。5例耐药个体中有1例出现了对蛋白酶抑制剂(PIs)类药物的耐药突变、3例出现了对核苷类逆转录酶抑制剂（NRTIs）、2例出现了对非核苷类逆转录酶抑制剂（NNRTIs）、1例出现了同时对NRTIs、NNRTIs类药物的耐药突变。 结论： 山东省新报告未治疗的HIV-1感染者亚型以CRF01_AE为主，出现了对3类艾滋病抗病毒药物的耐药毒株，耐药传播处于低、中度流行水平，应引起高度重视。.
Keywords: Antiretroviral therapy; Drug resistance threshold survey; HIV; Sequence analysis.