张巍巍, 李颖, 王群, 赵林, 冯宝立, 蒋会婷, 郝帅, 李清, 张颖, 马红梅, 逄波. 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[J]. 疾病监测, 2020, 35(6): 518-522. DOI: 10.3784/j.issn.1003-9961.2020.06.013
引用本文: 张巍巍, 李颖, 王群, 赵林, 冯宝立, 蒋会婷, 郝帅, 李清, 张颖, 马红梅, 逄波. 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[J]. 疾病监测, 2020, 35(6): 518-522. DOI: 10.3784/j.issn.1003-9961.2020.06.013
Weiwei Zhang, Ying Li, Qun Wang, Lin Zhao, Baoli Feng, Huiting Jiang, Shuai Hao, Qing Li, Ying Zhang, Hongmei Ma, Bo Pang. Application of whole genome sequencing in etiology investigation of an outbreak of polyclonal Vibrio parahaemolyticus infection[J]. Disease Surveillance, 2020, 35(6): 518-522. DOI: 10.3784/j.issn.1003-9961.2020.06.013
Citation: Weiwei Zhang, Ying Li, Qun Wang, Lin Zhao, Baoli Feng, Huiting Jiang, Shuai Hao, Qing Li, Ying Zhang, Hongmei Ma, Bo Pang. Application of whole genome sequencing in etiology investigation of an outbreak of polyclonal Vibrio parahaemolyticus infection[J]. Disease Surveillance, 2020, 35(6): 518-522. DOI: 10.3784/j.issn.1003-9961.2020.06.013

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

  • 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。

     

    Abstract:
    ObjectiveApplying real-time PCR and whole genome sequencing in the etiology investigation of an outbreak caused by Vibrio parahaemolyticus.
    MethodsDiarrhea 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.
    ResultsA total of 13 V. parahaemolyticus strains were isolated from 9 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.
    ConclusionThis outbreak was caused by polyclonal V. parahaemolyticus. Whole genome sequencing has a good application prospect in pathogen analysis of diarrhea outbreak.

     

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