Objective To investigate the serotypes, virulence genes, antimicrobial resistance and molecular characteristics of Vibrio parahaemolyticus in Huzhou of Zhejiang Province. Methods Isolates of 92 V. parahaemolyticus in Huzhou were analyzed by serological typing, virulence gene detection, antibiotic resistance testing, and pulsed-field gel electrophoresis (PFGE) for molecular typing. Results O3∶K6 was the main serotype and tlh⁺tdh⁺trh- was the most frequently detected virulence genotype in clinical strains. O2∶Kut was the main serotype and tlh⁺tdh-trh- was the most frequently detected virulence genotype in environmental strains. Antibiotic resistance testing indicated that the isolates were highly resistant to ampicillin (90.22%,83/92), followed by gentamicin and tetracycline. Following the restriction enzyme Not I digestion, the 91 strains yielded 81 PFGE patterns, and 16 clones had similarity values of >85.00%. Conclusion The etiologic characteristics of V. parahaemolyticus varied, which might be one of the reasons for high incidence of foodborne disease in Huzhou. There may be cross-contamination between freshwater and seawater products, so it is necessary to strengthen supervision of food processing.