食品来源副溶血性弧菌毒力基因分布及分子分型研究

陈晶; 黄建飞; 刘丛丛; 刘灵燕; 赖小红; 杨国武; 兰全学

doi:10.13386/j.issn1002-0306.2016.06.041

食品来源副溶血性弧菌毒力基因分布及分子分型研究

Study on virulence genes distribution and molecular subtyping of Vibrio parahaemolyticus from different food resources

摘要

摘要: 分析食品中分离获得的副溶血性弧菌主要毒力基因的分布特征、血清型及分子分型特点,为食源性副溶血性弧菌感染暴发的早期预警和疫情溯源提供实验室数据。提取40株副溶血性弧菌基因组,分别用PCR检测VcrD1、Vp1680、VopP和VcrD2基因,荧光PCR检测tdh、trh和tlh基因。对40株菌株进行血清型分型并采用脉冲场凝胶电泳（PFGE）对其进行分子分型。利用BioNumerics软件对图谱进行聚类分析,建立PFGE分子分型数据库。40株菌株均未检测到tdh、trh、Vop P和VcrD2基因,tlh、VcrD1和Vp1680基因阳性检测率为100%。40株菌株中含有16种血清型,其中9株未分型,主要血清型为O₂∶K₂₈和O₅∶K₁₇,血清型分布呈多样性。40株菌株共获得39个不同的PFGE带型,PFGE呈现遗传多样性,无优势带型。本实验食品环境中分离的副溶血性弧菌未发现毒力因子,副溶血性弧菌血清型呈分散趋势且菌株之间亲缘关系较远。

Abstract: The distribution of virulence genes、serogroup and molecular subtyping of Vibrio parahaemlyticus isolated from different resources of food were analysed,which contribute to provide scientific evidence for surveillance and source tracking. Forty strains of Vibrio parahaemolyticus genomes were extracted,VcrD1,Vp1680,Vop P,VcrD2 were amplified by PCR and tdh,trh,tlh genes were detected by Real-time fluorescent PCR. All of strains were subtyped by pulsed-field gel electrophoresis and serotyped. PFGE patterns were clustered by Bionumerics software. Virulence genes tdh,trh,Vop P and Vcr D2 were not detected,genes of tlh,VcrD1 and Vp1680 were all positive detected. Sixteen serotypes were observed,the serotyping of 9 isolates failed. The major were O₂∶K₂₈ and O₅∶K₁₇. No predominant serotype was detected. Thirty-nine PFGE patterns were detected and no predominant pattern. No virulence genes were detected. The isolates from food resource showed diversified serotype feature and far genetic distance.