Objective To establish a novel method to specifically detect Vibrio parahaemolyticus toxR and tdh genes in food by dual-color fluorescent real-time PCR. Methods The primers and probes targeting the toxR (trans-membrane transcription activator) and tdh (thermostable direct hemolysin) were designed, and their specificity and sensitivity were tested with the Taqman probe dual-color fluorescent real-time PCR amplification system. The optimized method was used to detect the isolated bacterial strains and to explore the distribution of toxR and tdh genes. Results Amplification curves of both toxR and tdh genes were obtained from standard V. parahaemolyticus strains and the three V. parahaemolyticus strains isolated from food-poisoning patients. How-ever, no such amplification curve was observed for all the other newly isolated 31 bacterial strains, including Vibrio alginolyticus and L. monocytogenes from the genus Vibrio and the family Enterobacteriaceae. All the 37 food V. parahaemolyticus strains did not carry the tdh virulence gene. Furthermore, the detection sensitivity of this novel method reached 3.6×102 cfu/mL. Conclusion The novel dual-color fluorescent real-time PCR method was able to specifically and effectively detect V. parahaemolyticus in food samples.
标题：食品中副溶血性弧菌toxR和tdh基因双色荧光PCR检测方法的建立
英文标题：Detection of Vibrio parahaemolyticus toxR and tdh genes in foods by dual-color fluorescent real-time PCR
