我国主要沿海地区O1群霍乱弧菌分子分型特征分析  

Analysis of molecular typing characteristics of Vibrio cholerae O1 in major coastal areas of China

作　　者：柳婷婷 李萍 杨文辉 金大智 高姗[2] 王菁 王景林 康琳 辛文文 LIU Ting-ting;LI Ping;YANG Wen-hui;JIN Da-zhi;GAO Shan;WANG Jing;WANG Jing-lin;KANG Lin;XIN Wen-wen(College of Life Sciences,Hebei Normal University,Hebei Key Laboratory of Animal Physiology,Biochemistry and Molecular Biology,Shijiazhuang 050024,China;State Key Laboratory of Pathogen and Biosecurity,Beijing Institute of Microbiology and Epidemiology,Academy of Military Medical Sciences(AMMS),Beijing 100071,China;Hangzhou Medical College,Hangzhou 310059,China;Zhejiang Provincial Center for Disease Control and Prevention,Hangzhou 310057,China)

机构地区：[1]河北师范大学生命科学学院,河北省动物生理生化与分子生物学重点实验室,石家庄050024 [2]军事科学院军事医学研究院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071 [3]杭州医学院,杭州310059 [4]浙江省疾病预防控制中心,杭州310057

出　　处：《中国人兽共患病学报》2022年第1期10-18,共9页Chinese Journal of Zoonoses

摘　　要：目的采用MLST方法及基质辅助激光解析电离飞行时间质谱(Matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF MS)技术研究我国主要沿海地区192株O1群霍乱弧菌的种群结构、进化趋势及流行特性。方法PCR扩增192株O1群霍乱弧菌的7个管家基因adk、gyrB、mdh、metE、pntA、purM和pyrC,根据核酸序列信息获得其等位基因型及序列型(Sequence Type,ST)。利用DnaSP 5.0及eBURST Version 3.0分析序列多样性,SplitsTree 4.0、STRUCTURE 2.2等软件分析霍乱弧菌种群结构特征,同时数据库中非O1/O139群对应的STs也纳入分析,以比较其与O1/O139群对应STs在克隆群和种群结构方面的关系。此外还应用BioTyper软件,对霍乱弧菌蛋白指纹图谱数据进行聚类分析。结果192株O1群霍乱弧菌共22个序列型,其中11个为本研究新发现的序列型;ST189、ST191和ST184存在2个不同组别间的重组现象。SplitsTree 4.0种群结构分析结果显示,22个序列型共分成2个分支,STRUCTURE种群结构分析将22个序列型分为3个亚群,不同的16S rDNA序列存在共有的ST,如ST69、ST75和ST173;非O1/O139群对应的STs大部分以单体形式存在,仅形成4个克隆群,在种群结构分析中大部分STs与O1/O139群对应的STs划分到同一个亚群中;MALDI-TOF MS聚类分析结果显示,当距离为200时,22个STs被分为两组,当距离为100时,被分成3组;霍乱弧菌整体的重组率(包括O1/O139群和非O1/O139群)要远远高于O1/O139群的重组率。结论192株O1群霍乱弧菌MLST分析表明ST69和ST75是O1群霍乱弧菌最主要的2个序列型。7个管家基因重组/突变的比值差异较大,但整体而言,重组比突变更容易发生。O1/O139群和非O1/O139群霍乱弧菌间仍存在较大差异。霍乱弧菌MLST分型与基于蛋白指纹图谱的分型结果间尚未发现明显的联系。To research the population structure,evolutionary trends,and epidemic characteristics of 192 strains of the Vibrio cholerae O1 group,we performed MLST and matrix-assisted laser desorption/ionization-time of flight mass spectrometry(MALDI-TOF MS)in this study.The seven housekeeping genes adk,gyrB,mdh,metE,pntA,purM,and pyrC from 192 strains of V.cholerae O1 were amplified by PCR,and their allelic genotypes and sequence types(ST)were obtained on the basis of the nucleic acid sequence information.DnaSP 5.0 and eBURST Version 3.0 were used to analyze sequence diversity,and SplitsTree 4.0 and STRUCTURE 2.2 were used to analyze the structural characteristics of V.cholerae populations.The STs corresponding to the non-O1/O139 group in the database were also included in the analysis to compare their relationship with the STs corresponding to the O1/O139 group in terms of clonal group and population structure.In addition,Biotyper software was used for cluster analysis of the V.cholerae protein fingerprint data.Finally,we identified 22 STs in 192 strains of V.cholerae O1,11 of which were newly discovered.ST189,ST191,and ST184 showed recombination between two groups.Population structure analysis with SplitsTree 4.0 revealed that the 22 sequence types were divided into two branches,and population structure analysis with STRUCTURE 2.2 divided the 22 sequence types into three subgroups.Different 16 S rDNA sequences had shared STs,such as ST69,ST75,and ST173.Most of the STs corresponding to the non-O1/O139 group existed in the form of monomers,and only four clonal groups formed.In population structure analysis,most of the STs and the STs corresponding to the O1/O139 group were divided into the same subgroup.The results of cluster analysis with MALDI-TOF MS indicated that when the distance was 200,the 22 STs were divided into two groups,and when the distance was 100,they were divided into three groups.The overall recombination rate of V.cholerae(including the O1/O139 group and non-O1/O139 group)was much higher than that of the O1

关 键 词：霍乱弧菌 O1血清群 多位点序列分型 基质辅助激光解析电离飞行时间质谱 

分 类 号：R378.3[医药卫生—病原生物学]