全基因组数据应用于多克隆副溶血弧菌感染暴发的病原学分析  

Application of whole genome sequencing in etiology investigation of an outbreak of polyclonal Vibrio parahaemolyticus infection

作　　者：张巍巍 李颖[2] 王群 赵林 冯宝立 蒋会婷 郝帅 李清 张颖 马红梅[2] 逄波[3,5] Zhang Weiwei;Li Ying;Wang Qun;Zhao Lin;Feng Baoli;Jiang Huiting;Hao Shuai;Li Qing;Zhang Ying;Ma Hongmei;Pang Bo(Miyun District Center for Disease Control and Prevention,Beijing 101500,China;Shunyi District Center for Disease Control and Prevention,Beijing 101300,China;National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Department of Laboratory Medicine,Renji Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 200127,China;State Key Laboratory of Infectious Disease Prevention and Control,Beijing 102206,China)

机构地区：[1]北京市密云区疾病预防控制中心,北京101500 [2]北京市顺义区疾病预防控制中心,北京101300 [3]中国疾病预防控制中心传染病预防控制所,北京102206 [4]上海交通大学医学院附属仁济医院,上海200127 [5]传染病预防控制国家重点实验室,北京102206

出　　处：《疾病监测》2020年第6期518-522,共5页Disease Surveillance

摘　　要：目的应用实时荧光PCR方法和基因组重测序分析等技术对一起副溶血弧菌(VP)导致的腹泻暴发事件进行病原学分析。方法收集暴发事件的病例信息,采集病例样本,使用实时荧光PCR方法筛选常见腹泻致病菌,分离鉴定VP并进行血清型鉴定,检测菌株毒力基因,采用微量肉汤稀释法检测菌株对抗菌药物的敏感性,使用全基因组测序技术进行菌株的遗传学分析。结果9份肛拭子共分离出13株VP菌株,其中O4∶KUT血清型7株(trh-/tdh+),O1∶KUT血清型6株(5株trh+/tdh+,1株trh-/tdh-)。在检测的30种抗菌药物中,所有菌株仅对氨苄西林耐药,对其他29种抗菌药物均敏感。在基于全基因组序列的遗传学分析中,13株VP形成4个相互独立且遗传距离较远的分支,Lineage 1(O4∶KUT、trh-/tdh+,2株)、Lineage 2(O1∶KUT、trh-/tdh-,1株)、Lineage 3(O1∶KUT、trh+/tdh+,5株)和Lineage 4(O4∶KUT、trh-/tdh+,5株),其中3例患者由来源于3个不同遗传分支的VP混合感染。结论本次事件由多血清型、多克隆的VP感染导致,基因组重测序技术在腹泻病暴发的病原分析中有较好的应用前景。Objective Applying real-time PCR and whole genome sequencing in the etiology investigation of an outbreak caused by Vibrio parahaemolyticus.Methods Diarrhea patient stool samples and epidemiological data were collected.Suspected pathogens were screened with real-time PCR.V.parahaemolyticus strains were isolated,identified and their serotypes were confirmed by agglutination with commercial diagnostic serum.The existences of tdh and trh genes were identified with real-time PCR.The drug resistance of the strains was determined by broth micro-dilution method,and whole genome sequencing data was used for genetic analysis.Results A total of 13 V.parahaemolyticus strains were isolated from9 anal swabs of patients,including 7 strains of O4∶KUT(trh-/tdh+)and 6 trains of O1∶KUT(5 trh+/tdh+strains,1 trh-/tdh-strain).All the V.parahaemolyticus strains were only resistant to ampicillin and sensitive to other 29 kinds of antibiotics.The 13 V.parahaemolyticus strains formed four independent and distant related lineages,Lineage 1(O4∶KUT,trh-/tdh+,2 strains),Lineage 2(trh-/tdh-,1 strain),Lineage 3(O1∶KUT trh+/tdh+,5 strains)and Lineage 4(O4∶KUT,trh-/tdh+,5 strains).Three patients had mixed infections of V.parahaemolyticus from three different genetic branches.Conclusion This outbreak was caused by polyclonal V.parahaemolyticus.Whole genome sequencing has a good application prospect in pathogen analysis of diarrhea outbreak.

关 键 词：副溶血弧菌 暴发 多克隆 基因组重测序 耐热相关溶血素基因 

分 类 号：R378[医药卫生—病原生物学]