我国东部沿海地区水产品中副溶血弧菌分离株的基于碱基序列和肽链序列的多位点序列分型研究

发布时间：2018-02-17 06:05

本文关键词： 副溶血弧菌基于碱基序列的多位点序列分型基于肽链的多位点序列分型克隆复合体　出处：《军事医学》2015年11期 　论文类型：期刊论文

【摘要】：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。
[Abstract]:Objective to investigate the virulence gene carriers and the relationship between virulent genes and clone populations of Vibrio parahaemolyticus isolated from aquatic products in the eastern coastal areas of China from 2007 to 2012. Methods from Guangdong Province, Fujian Province, Shanghai Province, Jiangsu Province during the period of 2007 ~ 2012, the virulence genes of Vibrio parahaemolyticus strains were collected. Vibrio parahaemolyticus isolated from aquatic products in Beijing, Shandong Province, A total of 79 strains of Vibrio parahaemolyticus were identified by fluorescence PCR based on tlh gene. The virulence gene, heat-resistant direct hemolysin, heat-resistant direct hemolysin (HAL) and the marker gene orf8 of epidemic group were detected by fluorescence PCR method. The subgroup analysis and clone complex analysis were carried out by using the internationally recommended multilocus sequence typing (MLST) method, and the minimal spanning tree based on St and p St was constructed. The results of cloning complex analysis based on e BURST showed that the carrying rate of tdh positive orf8 gene in 79 aquatic product isolates were 8.86% and 8.86% respectively. These 79 isolates belonged to 69 St types and 3 grams respectively. Carina complex, 62 monomers. In multiposition sequence typing based on peptide chain, The 79 isolates of Vibrio parahaemolyticus could be divided into 23 pST types, including 1 clone complex and 2 monomers. Conclusion Vibrio parahaemolyticus isolated from aquatic products in the eastern coastal areas of China is highly polymorphic, but the virulence gene carrying rate is relatively low. ST3 type is the main type of polymorphic AA-MLST typing.
【作者单位】：北京工商大学食品学院食品添加剂与配料北京高校工程研究中心;中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室;
【基金】：国家自然科学基金资助项目(31400161,81401715) 北京工商大学青年教师科研启动基金资助项目(QNJJ2014-25)
【分类号】：R155.5

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