辽宁省艾滋病治疗患者耐药基因特征及分子流行病学分析

HIV-1 drug-resistance profiles of treated AIDS patients in Liaoning： genetic characteristics and prevalence

目的了解辽宁省自2004年以来抗病毒治疗的艾滋病患者中基因耐药突变和HIV-1耐药株的流行率。方法选取2004—2008年全省接受一线抗病毒治疗艾滋病患者104个样本，病毒载量检测，高于最低检测限有12个样本，经HIV蛋白酶（PR）全长和部分反转录酶（RT）基因区RT-PCR扩增、测序，运用Contig Express1对序列拼接、编辑和校正，上传Stanford HIVDrugResistance Database（http：／／hivdb．stanford．edu），在线耐药序列分析，寻找耐药位点，评价目前使用的一线药物的治疗效果和耐药株流行趋势。结果104份治疗患者的样本中，92份病毒载量检测已经小于最低检测限，治疗有效率88．4％（92／104）。余下的12份样本经RT-PCR、巢式PCR和5个序列的正反方向测序，8份样本得到PCR有效扩增和测序（均获得5条有效测序序列）。RT区序列检测，8份样本中有5份存在核苷类反转录酶抑制剂（NRTIs）相关突变，耐药突变率为4．81％（5／104），M184V出现较早（治疗8个月）；6份存在非核苷类反转录酶抑制剂（NNRTIs）相关突变，耐药突变率为5．77％（6／104），以K103N、Y181C出现频次较高，使NNTRIs存在高度的多重交叉耐药。未发现蛋白酶抑制剂主要耐药相关突变，总耐药突变率为6．73％（7／104）。结论辽宁省艾滋病治疗患者中耐药突变仍处于低流行状态，与国内其他地区原发耐药流行率接近。目前使用的一线反转录酶抑制剂药物仍有效。但同时怎样解决NNRTIs突变导致的多重耐药、调整药物配伍、提高用药依从性、跟踪分析病例个案发展是今后工作努力的方向。

Objective Since the advent in 2004 of highly active antiretroviral therapy （HAART） in Liaoning, a dramatic improvement had been seen in the number of patients attaining undetectable viral loads （92/104）, but the extent of mutation diversity on human immunodeficiency virus 1 （HIV-1） and the prevalence of drug resistance had remained elusive. This study aimed to analyze both HIV-1 mutation profiles and prevalence related to antiretroviral resistance following therapeutic failure. Methods A total of 104 blood samples circling Liaoning from HAART-treated between 2004 and 2008 were studied. Patients＇ CD4＋ T-cell count and viral load were determined. HIV-1 pol （PR and part of RT） gene fragments were amplified from patients＇ plasma by reverse transcriptase polymerase chain reaction （RT-PCR） and nest-PCR, subsequently sequenced and analyzed. Results CD4＋ T cell numbers and viral replication capacity were assessed. 88.4% （92/104） of the patients were successful after initial non-suppressive NRTI ＆ NNRTI-based HAART regimens. Subjects on non-nucleoside reverse transcriptase inhibitor （NNRTI） regimens developed more （6/104） drug-resistance mutations than those on nucleoside reverse transcriptase inhibitor （NRTI） regimens did （5/104）. No protease-inhibitor （PI） drug resistance mutations developed. The whole rate of drug resistance mutations was about 6.73%. Subjects developing NNRTI-resistance （NNRTI-R） seemed more likely to develop drug-resistant viremia than with NRTI-based HAART. Conclusion This finding might have implications in which that the prevalence of drug-resistance mutations was low but remained risk of transmission in HIV-infected therapeutic failure. Meanwhile, data from the present study showed that there was a high frequency of primary mutations, which offered resistance to nrti and nnrti. Monitoring patients with treatment failure seems an important tool in helping the physicians to improve their treatment schedule and to carry out epidemiological surveillance programs.