Genetic characterization on near full-length genomes of two HIV-1 unique recombinant forms CRF55_01B/CRF07_BC in Beijing
肖明凤 1李佳 2李茜瑶 2刘安 3孙丽君 3卢红艳 2辛若雷2
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作者信息
1. 中国医科大学公共卫生学院,沈阳 110122
2. 北京市疾病预防控制中心性病艾滋病防治所 100013
3. 首都医科大学附属北京佑安医院感染中心门诊 100069
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摘要
目的 对北京治疗前耐药监测发现的具有遗传相似性的两株新发重组毒株进行前病毒HIV-1 DNA近全长基因组扩增和序列分析,以了解其遗传特征和重组模式.方法 对北京佑安医院募集的2例新发重组毒株感染者,用广州海力特公司核酸提取和纯化试剂从白细胞富集层提取前病毒HIV-1 DNA.运用SimPlot 3.5软件分析重组模式和断点,并用Mega 11构建分片段最大似然法进化树验证重组事件.结果 从2个男男性行为(men who have sex with men,MSM)感染者扩增获得HIV-1近全长基因组序列.SimPlot重组断点分析显示BL6218-00和BL6017-01基因组均以CRF55_01B为骨架,分别在gag和env基因区,或gag、pol、vpu/env、nef/3'LTR基因区,由CRF07_BC重组而成.2条序列的CRF07_BC片段均与来自MSM人群的CRF07_BC_N参考毒株序列聚集成单系进化簇.结论 用前病毒DNA扩增方法从北京MSM人群发现了两株以CRF55_01B为骨架且重组模式不同的独特重组型毒株,重组亲本CRF07_BC均来自我国北方MSM人群.
Abstract
Objective To perform near full-length HIV-1 genome amplification of two novel recombinant strains with genetic similarity identified by pret-reatment drug resistance surveillance in Beijing,and investigate the genetic characteristics and recombinant modes.Methods Two individuals infected by novel recombinant strains were recruited from Beijing You'an Hospital.The proviral DNA was extracted from the leukocyte enrichment layer using the Nucleic Acid Extraction and Purification Reagent from Supbio.Recombination breakpoints were analyzed using SimPlot 3.5 software.Segmental maximum likelihood trees were constructed by Mega 11 to verify the recombination events.Results Two near full-length genomes(BL6218-00 and BL6017-01)were obtained by nested PCR amplification of two samples from men who have sex with men(MSM).SimPlot analyses revealed that the two cases shared the backbone of CRF55_01B,with the substituted CRF07_BC fragments in the gag and env gene regions,or the gag,pol,vpu/env,nefl3'LTR gene regions,respectively.The segmental evolutionary analyses showed that the CRF07_BC fragments of BL6218-00 and BL6017-01 were merged into a monophyletic cluster with CRF07_BC_N reference strains.Conclusions Two unique recombinant forms,with CRF55_01B as backbone and two modes of recombination,were identified from from the MSM population in Beijing using of proviral DNA amplification.The recombination parents of CRF07_BC genetically originated from the strains circulating among the MSM population in northern China.
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