In this paper, two selective media—TCBS agar and CHROMager—were utilized for the isolation of Vibrio parahaemolyticus (Vp). According to the biochemical identification, all the 27 suspected strains which were isolated from 60 samples of seafood were identified to be Vp, 22 strains of them were from fresh seafoods and 5 strains were from frozen ones. With further research on 4 special biochemical tests, comparing these 27 Vp strains with 32 Vp clinic strains isolated from patients, there were some difference in Hemolysis test, Urease test and Citrate test, while no significant difference in Arabinose test. As such, it supplies a theoretic basis for studying in biochemical characteristics of Vp from different sources.