摘要
目的通过分析云南省一株HIV-1独特重组毒株的基因结构和重组特点了解云南省独特重组毒株的演变情况。方法对云南省艾滋病关爱中心抗病毒治疗的耐药患者进行耐药监测后,从耐药监测序列库中200条序列进行全长扩增,扩增成功后选取具有代表性的独特二代重组毒株进行研究。采集血浆500μL,分离血浆并提取纯化血浆中的RNA。全长序列分为三个部分,运用RT-PCR的方法分别扩增该病毒基因的3'半分子,gag区和pol区,扩增产物测序后获得该毒株近全长基因序列。用Genotyping、BLAST、RIP和jpHMM在线分析工具和Simplot3.5软件进行重组分析。结果获得8850 bp近全长HIV-1基因序列,重组分析显示,该条序列是以C亚型为骨架,B(B')和C亚型作为片段插入的一条位置重组毒株,该条序列共有10个重组断点将序列分为11个片段,其中5个为C片段,4个B片段,2个CRF01_AE片段。结论云南省HIV-1URF的多样性不断演变,尤其是以C、B(B')和CRF01_AE亚型的二代重组,需要持续监测,以应对诊断、抗逆转录病毒治疗和预防方面日益增加的挑战。
Objective To understand the evolution of a unique recombinant strain of HIV-1 in Yunnan Province by analyzing the gene structure and recombination characteristics of a unique recombinant strain of HIV-1 in Yunnan Province.Methods After drug resistance monitoring of drug-resistant patients treated with antiviral therapy in Yunnan AIDS Care Center,200 sequences from the drug resistance monitoring sequence library were amplified in full length,and representative unique second-generation recombinant strains were selected for the study after successful amplification.Plasma(500μL)was collected and separated,and then RNA was extracted from the purified plasma.The full-length sequence was divided into three parts,and the 3'half molecule,gag region and pol region of the virus gene were amplified by RT-PCR,respectively.The near-full-length gene sequence of the strain was obtained after sequencing of the amplified product.Recombination analysis was performed with Genotyping,BLAST,RIP and jpHMM online analysis tools and Simplot3.5 software.Results The 8850 bp near full-length HIV-1 gene sequence was obtained.Recombination analysis showed that the sequence was a recombinant strain with subtype C as the backbone and subtypes B(B')and C as a fragment insertion position.There were 10 recombination breakpoints in this sequence.The sequence was divided into 11 fragments,of which 5 were C fragments,4 B fragments,and 2 CRF01_AE fragments.Conclusion The diversity of HIV-1 URF in Yunnan Province continue to evolve,especially the second-generation recombination of C,B(B')and CRF01_AE subtypes.Our ongoing surveillance is needed to meet the increasing challenges in diagnosis,antiretroviral therapy,and prevention.
作者
周熙姝
张米
刘家法
王佳丽
杨壁珲
李健健
董兴齐
ZHOU Xishu;ZHANG Mi;LIU Jiafa;WANG Jiali;YANG Bihui;LI Jianjian;DONG Xingqi(Yunnan Provincal Infectious Diseases Hospital/AIDS Care Center Laboratory,Kunming 650301,China)
出处
《中国皮肤性病学杂志》
CAS
CSCD
北大核心
2022年第8期898-904,共7页
The Chinese Journal of Dermatovenereology
基金
云南省高层次卫生技术人才培养经费资助项目(H-2018050)
云南省艾滋病病毒学及临床诊疗技术创新研究中心(202102AA310005)
国家自然基金项目(81960605)
云南省教育厅科学研究基金项目(2019Y0260)。
关键词
人类免疫缺陷病毒-1
独特重组型
近全长基因序列
Human immunodeficiency virus type 1
Unique recombinant form
Near-full-length gene
