Establishment of Digital PCR Assay for Detection of Vibrio parahemolyticus

Objective:To establish a quantitative accuracy method to detect Vibrio parahemolyticus based on droplet digital PCR. Methods:With the specific sets of primers and probe synthesized to target genes including TLH genes, specificity and sensitivity were tested based on ddPCR and quantitative accuracy analysis was performed on the results. In order to verify the feasibility of the method, positive strain was added to seafood in which was often detected Vibrio parahaemolyticus.Conclusion:Vibrio parahaemolyticus performed good specificity in ddPCR detection in this experiment. The range of effective DNA concentration was 2~19440 copies/20μL. Bacterial suspension concentration was 50~4.86×10⁵CFU/mL, which had no significant difference with 3M test (p>0.05). Compared with real-time PCR, the concentration limit of detection was enhanced by two orders of magnitude. The results showed that quantitative detection of Vibrio parahaemolyticus by ddPCR technology had good specificity, high sensitivity and accuracy, which had practical application value.