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目的对引起一起食物中毒的副溶血性弧菌进行实验室鉴定,分析菌株间的相关性。方法按照国标GB4789和有关规范对采自患者的粪便肛拭标本及砧板涂抹标本进行肠道致病菌检测,同时采用实时荧光PCR对耐热直接溶血素(TDH)毒力基因进行鉴定。提取副溶血性弧菌分离株基因组DNA,经限制性内切酶SfiⅠ酶切后进行脉冲场凝胶电泳(PFGE),获得指纹图谱,利用BioNumerics6.6软件进行聚类分析。结果从病人和砧板标本共分离出8株O1血清型副溶血性弧菌,其TDH毒力基因为阳性。聚类分析显示,分离自中毒病人和砧板的8株副溶血性弧菌的指纹图谱相似性高达100%。结论引起该起食物中毒的病原菌为携带TDH毒力基因的O1血清型副溶血性弧菌,且来自同一污染源。
Objective To identify the strains of Vibrio parahaemolyticus that cause food poisoning and analyze the correlation between strains. Methods According to the national standard GB4789 and the relevant norms, the stool samples collected from the patients and the samples of the cutting board were tested for intestinal pathogenic bacteria. At the same time, the real-time fluorescent PCR was used to identify the virulence genes of TDH. The genomic DNA of Vibrio parahaemolyticus was extracted and digested with restriction endonuclease Sfi I and then subjected to PFGE. The fingerprint of Vibrio parahaemolyticus was analyzed by BioNumerics6.6 software. Results Eight strains of O1 serotype Vibrio parahaemolyticus were isolated from the patient and the anvil samples, and the TDH virulence gene was positive. Cluster analysis showed that the fingerprints of eight strains of Vibrio parahaemolyticus isolated from poisoned patients and chopping boards were as similar as 100%. Conclusion The pathogen causing the food poisoning is O1 serotype Vibrio parahaemolyticus carrying TDH virulence genes, and it is from the same source of contamination.