The aim of this research was to establish a novel method based on tdh1 gene variation for identifying V.parahaemolyticus pandemic strains.Methods tdh sequences from various strains were compared, and specific sites for pandemic strains were searched. Allele-specific PCR was established based on the selected site, and verified by known isolates and isolates from 2014, compared with the joint of GS-PCR and tdh method as a reference methods. Results 26 sites in 6 tdh sequences from 3 isolates showed polymorphism, and only the 368^th base could be used to identify pandemic strains. tdh1_368 allele-specific PCR was based on this site which could discriminate pandemic O3∶K6 strains before and after 1996. This method could also discriminate other serovariants of pandemic strains from non-pandemic strains. According to the detection of 1 067 isolates from 2014, complete consistent was showed between the novel method and the reference method.Conclusion tdh1_368 allele-specific PCR was useful for V.parahaemolyticus pandemic strains identification, and the detection target was more simple and direct compared with the methods reported. It was the first time to use this mutation for pandemic strains identification.