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目的研究受污染海产品、海产品养殖水及食物中毒临床分离的副溶血性弧菌的生化特性、种特异性基因片段检测方法的应用和毒素基因携带情况。方法66株副溶血性弧菌采用全自动微生物鉴定、MPN值定量、氯化物含量(离子色谱法)、R72H基因片段和tdh、trh毒素基因PCR检测等方法进行研究。结果定性定量分析方面,98.5%的菌株不发酵阿拉伯糖;牡蛎样本MPN值均为≥24000/100g,并检出2株携带trh毒素基因;23/30份养殖水检出副溶血性弧菌,其中海丰监测点6份阳性水样的氯化物含量均值仅为3160mg/L,显著低于实验室培养的30g/L的含量。基因检测方面,VITEK鉴定R72H基因片段结果与检测相符;13株食物中毒临床分离株均携带tdh毒素基因;6.7%的海产品分离株、15.4%食物中毒临床来源株同时携带两种毒素基因。结论通过从海产品和养殖水的检出情况以及分离株的生化特性、毒素基因资料,为副溶血性弧菌食物中毒溯源提供重要的数据源。
Objective To study the biochemical characteristics of Vibrio parahaemolyticus isolated from contaminated seafood, aquaculture water and food poisoning, the application of species-specific gene fragment detection and toxin gene transfer. Methods 66 strains of Vibrio parahaemolyticus were studied by means of automatic microbial identification, MPN value quantification, chloride content (ion chromatography), R72H gene fragment and tdh, trh toxin gene PCR. Results In qualitative and quantitative analysis, 98.5% strains did not ferment arabinose. The MPN values of oyster samples were all ≥24000 / 100g, and two strains carrying trh toxin genes were detected. 23/30 parts of culture water were used to detect Vibrio parahaemolyticus, Among them, the average chloride content of 6 positive samples in Haifeng monitoring station was only 3160mg / L, which was significantly lower than that in laboratory culture of 30g / L. In terms of genetic testing, the results of VITEK identification of R72H gene fragment were in good agreement with the detection results. All 13 clinical isolates of food poisoning carried tdh toxin gene, 6.7% of marine products and 15.4% of clinical isolates of food poisoning. Conclusion The detection of Vibrio parahaemolyticus food poisoning provides an important data source through the detection of seafood and culture water as well as biochemical characteristics of isolated strains, toxin gene data.