Detection of Vibrio parahaemolyticus by RPA Isothermal Amplification Technology

Objective:To establish a rapid RPA-exo fluorescent probe detection method for Vibrio parahaemolyticus. Methods:Using RPA technology and irgB as target gene to design RPA-exo primer and probe,the primers were combined and screened to establish a rapid method. Its specificity,sensitivity,simulated contamination experiment and practical application effect were tested. Results:The construction method results were obtained in 15 min,with high specificity and no cross-reaction.The sensitivity was 1.0×10³ CFU/mL,DNA detection limit was 0.35 pg/μL,plasmid detection limit was 1×10³ copies/μL. The added final concentration 1.36×10³ CFU/mL of Vibrio parahaemolyticus could be tested without enrichment culture in simulated contamination experiment. In the actual sample test,3 samples of 10 aquatic products were detected Vibrio parahaemolyticus. The results was consistent with the result of GB 4789.7-2013.Conclusions:A rapid RPA-exo method for the detection of Vibrio parahaemolyticus was successfully established in this study.