摘要目的：通过对65名HIV治疗队列中发生耐药的感染者进行较系统的前瞻性队列研究,阐明我国抗病毒治疗过程中,HIV RT蛋白耐药进化的结构限制性以及HIV耐药的发生规律。方法从2003年开始,每半年1次,连续4年对队列感染者进行随访采集静脉血样,提取病毒RNA后对RT基因进行RT-PCR扩增和测序。将所获得的序列进行系统进化、耐药位点变异和蛋白质结构分析。结果 RT酶中大部分的氨基酸位点都是保守的,而序列的变异仅发生在12.44%的位点之上。非核苷类逆转录酶抑制剂( NNRTI)高水平耐药位点多位于RT蛋白的内部,而中等水平耐药位点多位于蛋白质的表面。核苷类逆转录酶抑制剂( NRTI)高水平耐药位点均位于RT蛋白的表面,而低水平耐药位点均位于RT蛋白的内部。结论 NNRTI和NRTI类耐药突变位点在RT蛋白中截然相反的分布特征,可以为针对RT靶点的抗病毒药物的设计和开发起到一定的指导作用。

abstractsObjective To elucidate constraints on the evolution of HIV drug resistance from pro-tein structure and to analyze the characteristics of HIV drug resistance through a prospective cohort study among 65 HIV infectors receiving high active anti-retroviral therapy ( HARRT) . Methods 20 ml of periph-eral venous blood sample was collected from each subject every 6 months over a 4 year-follow up since 2003 and added into ethylene-diamine-tetra-acetic acid ( EDTA) tube. HIV RNAs were extracted from the blood samples. More than 400 sequences encoding the reverse transcripatase (RT) of HIV were obtained after RT-PCR amplification and sequencing analysis. The sequences were further analyzed for a better understanding of the molecular evolution, drug resistance mutations and 3D protein structure. Results Most amino acids sites were conservative in RT protein and only 12. 44% of the sites had mutations. Most high level mutations associated with resistance to non-nucleoside reverse transcriptase inhibitor ( NNRTIs) located at the core of protein, while medium level mutations associated with resistance to NNRTIs located at the surface of protein. Most high level mutations associated with resistance to nucleoside reverse transcriptase inhibitor ( NRTIs) lo-cated at the surface of protein, while low level mutations associated with resistance to NRTIs located at the core of protein. Conclusion Mutations that were associated with drug resistance to NRTIs and NNRTIs were occurred in different parts of HIV RT protein, which would be beneficial for the design and develop of ART drugs targeting RT.