The purpose of this study is to investigate the distribution of toxin genes and antimicrobial resistance of Vibrio parahaemolyticus isolated from seafood in Gwangju. A total of 335 seafood, including 163 shellfish, 97 fish, and 36 mollusk, were tested in this study. As a result, V. parahaemolyticus was detected in 123 (36.7%) of 335 seafood. The tdh gene was not detected in all strains, while the trh gene was detected in 3 strains (2.4%). According to antimicrobial susceptibility test, 116 strains (94.3%) represent resistance to ampicillin, and 1 strain (0.8%) represents resistance to trimethoprim/sulfametoxazole. However, all strains were sensitive to 9 antimicrobial agents, including amikacin, chloramphenicol, tetracycline, and more. Therefore, the risk of V. parahaemolyticus isolated from seafood in Gwangju is considered low, but continuous monitoring of V. parahaemolyticus in seafood is required.