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调查

Survey of HIV-1 drug resistance threshold in Shandong Province in 2013-2015

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CD4＋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.

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.