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目的了解2007~2012年来自我国东部沿海地区水产品中的副溶血弧菌分离株的毒力基因携带情况和菌株克隆群关系构成。方法收集2007~2012年间来自广东省、福建省、上海市、江苏省、山东省、北京市的水产品,分离自水产品中的副溶血弧菌株,经基于tlh基因荧光PCR鉴定为副溶血弧菌共79株。采用荧光PCR方法对其进行了毒力基因耐热直接溶血素(tdh)、耐热直接溶血素相关溶血素(trh)及流行组标识基因orf8检测,采用国际推荐的多基因座序列分型(MLST)方法对菌株进行亚群分析和克隆复合体分析,并构建基于ST型和p ST型的最小生成树,根据e BURST进行克隆群复合体分析。结果在79株水产品分离株中,tlh均为阳性,8.86%(7/79)的分离株tdh阳性,orf8基因的携带率为8.86%。此79株副溶血弧菌分离株分属于69个ST型,3个克隆群复合体,62个单体。在基于肽链的多位序列分型时,此79株副溶血弧菌分离株可分为23个p ST型,包括1个克隆群复合体,2个单体。去重组后剩余363个单核苷酸多态性(SNP)位点,以此363个SNP位点进行分型,可分为68个pattern型。结论来自我国东部沿海地区的水产品中分离的副溶血弧菌具有高度多态性,但毒力基因携带率较低,MLST分型时以ST3型为多,AA-MLST分型时以p ST1型为主。
Objective To understand the relationship between virulence genes of strains of Vibrio parahaemolyticus isolated from aquatic products in the eastern coastal areas of China from 2007 to 2012 and the relationship between bacterial clonal groups. Methods Aquatic products isolated from aquatic products from Guangdong, Fujian, Shanghai, Jiangsu, Shandong and Beijing from 2007 to 2012 were collected and identified as parahemolytic by fluorescent PCR based on tlh gene A total of 79 strains of Vibrio. The tdh, heat-resistant hemolysin-related hemolysin and orf8 gene were detected by fluorescence PCR, and sequenced by the internationally recommended multi-locus sequence MLST) was used to analyze the sub-population and clonal complex. The minimal spanning tree based on ST and p ST was constructed and the clonal complex was analyzed by e BURST. Results Among the 79 strains of aquatic products, tlh was positive, while 8.86% (7/79) of isolates were positive for tdh. The orf8 gene carrying rate was 8.86%. The 79 isolates of Vibrio parahaemolyticus belong to 69 ST type, 3 clonal complex and 62 monomer. In the multiple peptide sequencing-based sequencing, the 79 isolates of Vibrio parahaemolyticus were divided into 23 p ST types, including one clonal complex and two monomers. There were 363 single nucleotide polymorphisms (SNPs) remaining after the recombination, and 363 SNPs were classified into 68 patterns. Conclusion Vibrio parahaemolyticus isolated from aquatic products in the eastern coastal areas of China is highly polymorphic, but the virulence factor is low, with ST3 type in MLST typing and p ST1 in AA-MLST typing Based.