副溶血性弧菌引发食源性疾病的病原学检测及同源性分析

Pathogen detection of food-born diseases caused by Vibrio Parahaemolyticus infection and homologous analysis with pulsed field gel electrophoresis

目的对引起一起疑似食物中毒事件中的病原菌分离株进行病原学检测,并通过脉冲场凝胶电泳(PFGE)追溯致病菌的同源性。方法参照GB 4789.7-2013和相关规范对采回样本进行实时荧光定量PCR初筛、分离培养,以及生化鉴定、血清学分型、神奈川试验和药敏试验。使用脉冲场凝胶电泳(PFGE)技术分析致病菌的同源性。结果 14份样品中有5份检出副溶血性弧菌,血清学分型结果为O3:K6型,神奈川试验结果均为阳性。所检出的致病菌株对常见抗生素(阿米卡星、优立新、氨曲南、庆大霉素、环丙沙星、头孢类、呋喃坦啶、复方新诺明、哌拉西林、妥布霉素)敏感,而对氨苄西林耐药。PFGE分析结果提示其中4株样本菌株具有同源性,而另一株与其它菌株在分子水平上紧密相关。结论本事件是由副溶血性弧菌引发的群体性食物中毒,且致病菌来自同一污染源。

Objective To detect the pathogenic bacteria in cases with suspeceted food-born disease and analyse theirhomology with PFGE（Pulsed Field Gel Electrophoresis）. Methords The samples were preliminarily screened and thebacteia were cultured and isolated by real-time PCR in light on GB 4789.7-2013 and the relevant standard, biochemicalexamination, serum identification and Kanagawa experiment on the pathogenic Bacteria were carried out,and the drugresistance of the bacteria was tested by MIC. Then The homology of the pathogenic bacteria was expored using PFGEtechnology. Results Vibrio Parahaemolyticus serotype O3 and K6 were isolated from 5 anal swabs among the 14 samples andthe results of Kanagawa experiment were all positive. The pathogenic bacteria isolated from 5 suspected samples were sensitiveto common antibiotic drugs（ amikacin, unasyn,aztreonam,gentamycin,ciprofloxacin,cephalosporins,furantoin,SMZco,piperacillin,tobramycin）,while they were resistant to ampicillin. The PFGE pattern of 4 strains were the same, and the other one was closelyrelated to the strains at the molecular level indicated that the isolated pathogenic bacteria were closely and homologouslyrelated. Conclusion The food-born disease is caused by Vibrio Parahaemolyticus contamination and the pathogenic bacteriawas probable from a same pollution source.