Establishment of <I>Vibrio parahaemolyticus</I> detection by duplex TaqMan real-time PCR  assay

ObjectiveTo establish real-time PCR TaqMan assay for the detection of IVibrio parahaemolyticus/I and duplex real-time PCR TaqMan assay for the detection of toxic genes of IV. parahaemolyticus/I. MethodsThe conserved region of ItoxR/I gene of IV. parahaemolyticus/I was used to design primers and TaqMan probe, and the real-time PCR TaqMan system to detect IV. parahaemolyticus/I was established. The conserved region of Itdh/I and Itrh/I genes were used to design primers and probes, and the duplex real-time PCR TaqMan system to detect toxic genes of pathogenic IV. parahaemolyticus/I was established. The sensitivity and specificity of the assays to detect IV. parahaemolyticus/I was evaluated. ResultsThe sensitivity of the Real-time PCR TaqMan system to detect IV. parahaemolyticus/I was 10sup2 /supcopies/l; and the sensitivity of the duplex TaqMan Real-time PCR system to Itdh/I and Itrh/I was 10sup2 /supcopies/l. No amplification was observed in the templates of other bacterium when they were detected by the assay targeting ItoxR/I gene of IV. parahaemolyticus/I. Conclusion The established assays can detect IV. parahaemolyticus/I sensitively and specifically and determine toxic genes of pathogenic IV. parahaemolyticus/I, so it is a sensitive and specific way to detect IV. parahaemolyticus/I.