不同来源的副溶血性弧菌定性定量分析及毒素基因检测

Qualitative and Quantitative Analysis and Toxin Genes Detection of Vibrio parahaemolyticus Isolated from Different Resources

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 carry out the Vibrio parahaemolyticus isolated from marine products,cultured water and clinical patients by phenotyping qualitative and quantitative analysis combined with toxin genes detection. Method By means of VITEK identification,MPN methods,chlorine iron concentration （IC） analysis,R72H gene fragment and tdh、trh toxin genes detected by PCR. Results On the side of qualitative and quantitative results,Datum manifested that 98.5% of Vibrio parahaemolyticus isolates didn＇ t ferment arabinose, which differed from guidance of USFDA manual. Two isolates contained trh gene had been detected from Oyster that contaminated in high MPN （ ≥24 000/100 g）. Positive rates of cultured water were 23/30, 6/8 samples were positive whose means of concentration of chlorine iron were 3 160 mg/L, collected from certain region, were remarkably lower than concentration 30 g/L that cultured in vitro. On the other side of gene detection results, R72H gene fragment detection results VITEK identification results correspond with. Total 13 of isolations came from clinical resource contained tdh gene. Rates of which contained tdh, trh toxin genes of marine products, cultured water and clinical isolation were 6.7%, 0%, 15.4%, respectively. Conclusion Investigation of above characteristics of Vibrio parahaemolyticus isolated from marine products and cultured water taked a important role of tracing to the source of outbreaks and supplied database establishment with useful information.