2015年安徽地区副溶血性弧菌生物学特征及分子分型溯源分析

发布时间：2018-02-09 20:25

本文关键词： 副溶血性弧菌血清型毒力基因脉冲场凝胶电泳　出处：《中国病原生物学杂志》2017年04期 　论文类型：期刊论文

【摘要】：目的了解2015年安徽地区食品及腹泻病例粪便标本分离副溶血性弧菌的血清型及毒力基因特征,构建脉冲场凝胶电泳DNA指纹图谱库。方法采用玻片凝集法对分离自食品及腹泻病例的61株副溶血性弧菌进行血清学分型,采用Real-time PCR法检测tlh、tdh和trh毒力基因,通过脉冲场凝胶电泳(PFGE)对菌株基因组DNA进行分子分型,电泳图谱用BioNumerics 6.6软件进行聚类分析,并构建指纹图谱库。结果 61株副溶血性弧菌分为3个O群和20个K型,共23个血清型,以O3∶K6血清型为主,占26.2%(16/61)。61株副溶血性弧菌tlh、tdh和trh基因阳性率分别为100.0%(61/61)、29.5%(18/61)和0(0/61),病例分离株tdh基因阳性率为94.7%。61株菌株中有58株的基因组DNA得到有效酶切,分成50个PFGE带型,经BioNumerics6.6软件分析有5个相似系数大于80%的聚类群。其中病例株较为集中,多分布在A群和B群;食品株较为分散。病例株和食品株未聚集在同一群中。结论 2015年安徽地区分离的副溶血性弧菌有多种血清型,以O3∶K6型居多,病例分离株tdh基因携带率较高,通过构建PFGE图谱库可识别不同来源、不同区域、跨时间段菌株间的亲缘关系,为食源性疾病的溯源和暴发识别提供了依据。
[Abstract]:Objective to investigate the serotype and virulence gene characteristics of Vibrio parahaemolyticus isolated from fecal specimens of food and diarrhea cases in Anhui in 2015. Methods 61 strains of Vibrio parahaemolyticus isolated from food and diarrhea were serologically classified by slide agglutination method, and Ttlhtdh and trh virulence genes were detected by Real-time PCR. The genomic DNA of the strain was classified by pulsed field gel electrophoresis (PFGE). The electrophoretic pattern was analyzed by BioNumerics 6.6 and the fingerprint library was constructed. Results 61 strains of Vibrio parahaemolyticus were divided into 3 O groups and 20 K types. Of the 23 serotypes, O3: K6 serotype was the main type, accounting for 26.2R / 61n.61 strains of Vibrio parahaemolyticus Tl / TDH and trh gene positive rates were 100.00.61% 61R / 1861) and 0% / 61g / 1861, respectively. The positive rate of tdh gene was 94.771.61 strains of genomic DNA was effectively digested. It was divided into 50 PFGE bands, and there were five clusters with similarity coefficient greater than 80% by BioNumerics6.6 software, among which the case strains were concentrated and distributed in group A and group B. Food strains were scattered. Case strains and food strains were not clustered in the same group. Conclusion in 2015, Vibrio parahaemolyticus isolated in Anhui area had a variety of serotypes, most of which were O3: K6 type, and the tdh gene carrying rate of the case isolate strain was higher. The establishment of PFGE library can identify the relationship among strains from different sources, different regions and different time periods, which provides a basis for the tracing of foodborne diseases and the identification of outbreaks.
【作者单位】：安徽省疾病预防控制中心;
【基金】：国家科技重大专项(No.2013ZX10004203002004)
【分类号】：R378.3

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