目的 收集沧州市水产品中副溶血性弧菌的毒力基因和脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE)分型特征,为副溶血性弧菌的防控提供参考.方法 对2020-2022年1 056份海产品样本中分离出的139株副溶血性弧菌进行三重实时荧光聚合酶链式反应(polymerase chain reaction,PCR)技术检测本地区副溶血性弧菌的毒力基因;通过脉冲场凝胶电泳的基因图谱得到聚类图,采用BioNumerics 7.6软件对PFGE酶切片段聚类分析.分类变量采用卡方检验进行比较,双侧检验,以P＜0.05为差异有统计学意义.结果 1 056份样本检出139份副溶血弧菌阳性标本,检出率为13.16％,其中139株中有3株热稳定直接溶血素(thermostable direct hemolysin,tdh)基因阳性,具有毒力,带有毒力基因的副溶血性弧菌占比为2.16％.139株副溶血性弧菌部分同源,聚类分析相似度为36.5％～100.0％.结论 沧州地区带毒力的副溶血性弧菌主要为tdh毒力型,毒力基因分布特征与其他地区报道基本一致;PFGE图谱多态化,可有效应用于食源性疾病溯源分析及分子流行病学研究.为开展本地区副溶血性弧菌分布研究,预防副溶血性弧菌相关疾病的暴发打下基础.
Analysis of vibrio parahaemolyticus virulence genes and PFGE genotyping of aquatic products in Cangzhou from 2020 to 2022
Objective To collect the virulence genes and PFGE genotyping characteristics of Vibrio parahaemolyticus of aquatic products in Cangzhou City,to provide reference for prevention and control of Vibrio parahaemolyticus.Methods Triplex real-time PCR was performed on 139 Vibrio parahaemolyticus isolated from 1 056 seafood samples from 2020 to 2022 to detect the virulence genes of Vibrio parahaemolyticus in this region,the cluster map was obtained by PFGE gene map,BioNumerics 7.6 software was used for clustering analysis of PFGE.Categorical variables were compared by chi-square test,two-sided test,the difference P＜0.05 was considered statistically significant.Results Of the 1 056 Vibrio parahaemolyticus,139(13.16％)were positive for TDH gene,and 3(2.16％)of the 139 Vibrio parahaemolyticus were virulent.The similarity of 139 Vibrio parahaemolyticus was 36.5％-100.0％by cluster analysis.Conclusions The Vibrio parahaemolyticus in Cangzhou are mainly TDH virulence,and the distribution of virulence genes was similar to those reported in other regions,it can be effectively applied in the traceability analysis and Molecular epidemiology studies of food-borne diseases.The study laid the foundation for conducting regional Vibrio parahaemolyticus studies to prevent outbreaks of Vibrio parahaemolyticus diseases.
vibrio parahaemolyticusmolecular typingpulsed-field gel electrophoresiscluster mapvirulence genestriple real-time PCR
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