240株O3血清型副溶血性弧菌tdh和trh毒力基因研究

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目的 掌握不同来源O3血清型副溶血性弧菌毒力基因携带状况,分析毒力基因与传统神奈川现象的相关性,为深入研究副溶血性弧菌致病性提供基础数据.方法 用多重聚合酶链式反应(PCR)方法扩增tdh、trh毒力基因.按GB 4789.7-2013《食品安全国家标准食品微生物学检验副溶血性弧菌检验》方法进行神奈川试验.用x2检验方法确定tdh、trh基因与神奈川现象的相关性.结果 183株O3:K6血清型的副溶血性弧菌中,有182株检出tdh基因,1株检出trh基因,检出率分别为99.45% (182/183)和0.55% (1/183),神奈川试验阳性率为100.00%(183/183).57株O3非K6群血清型副溶血性弧菌tdh基因检出率为3.51% (2/57),未检出trh基因.神奈川试验结果阳性率为10.53% (6/57).经x2检验tdh基因与神奈川现象有相关性(x2=1.78,P>0.05),trh基因与神奈川现象无相关性(x2=186.01,P<0.05).结论 O3:K6血清型的副溶血性弧菌tdh基因携带率极高,且与神奈川现象相关性有统计学意义.神奈川现象可作为副溶血性弧菌tdh基因和致病性的指标.“,”Objective To analyze the relationship between virulence genes and traditional Kanagawa phenomena,based on the virulence genes carried by Vibrio parahaemolyticus isolates from different sources of serovar O3,to provide basic data for further studies on the pathogenicity of Vibrio parahaemolyticus.Methods Multiplex polymerase chain reaction (PCR) and Kanagawa phenomenon assay were performed to detect the existence of virulence associated genes tdh and trh as well as haematolysis of Vibrio parahaemolyticus serovar O3 strains with different origin.The correlation between tdh and trh genes and Kanagawa phenomenon were verified based on test.Results Among 183 Vibrio parahaemolyticus serovar O3:K6 strains,tdh gene was detected in 182 strains and trh gene exists in the remaining strain,detection rate of the two genes were 99.45% (182/183) and 0.55% (1/183),respectively.The positive rate of Kanagawa phenomenon was 100.00% (183/183).Among 57 Vibrio parahaemolyticus serovar O3 non-K6 strains,detection rate of tdh gene was 3.51% (2/57),trh gene was not detected.Furthermore,the positive rate of Kanagawa phenomenon was 10.53% (6/57).Significant correlation between tdh gene and Kanagawa phenomenon was verified (x2=1.78,P > 0.05),meanwhile,similar between trh gene and Kanagawa phenomenon was not detected (x2 =186.01,P <0.05).Conclusion Significant correlation between tdh gene and Kanagawa phenomenon was verified in Vibrio parahaemolyticus serovar O3:K6 strains,and pathogenicity of Vibrio parahaemolyticus can be detected by using PCR method rapidly.
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