To investigate and analyze a food poisoning with multiple pathogens including Vibrio parahaemolyticus (VP) and Proteus mirabilis.Methods Standardized questionnaire for food poisoning was used to collect information, descriptive epidemiology was used to detect the risk factors. The pathogens were isolated from the samples, enrichment was performed according to National Standard of Microbiological Examination of Food Hygiene, and the isolated VP were further identified by DNA fingerprint. Results Thirty diners were found and 5 lost, seventeen of which were consistent with case definition, and the incident rate was 56.67% (17/30). Diarrhea, abdominal pain especially epigastralgia was the main clinical symptom. Incubation period of the cases was 11-25 hours. No risk factor was found by cohort study. Nineteen strains were isolated from patients, of which were VP (9), Proteus mirabilis (8), Salmonellae (1) and C.jejuni (1). A strain of Proteus mirabilis was found in food handlers. A strain of VP and 4 strains of Proteus mirabilis were isolated from environmental swabs, the same situation was detected in food samples. Nine strains of VP isolated from patients were identified as the same genotype by molecular typing techniques, which genetic similarity reached more than 97%.Conclusion Considering both the investigation of field epidemiology, food hygiene and experimental results, it was a foodborne disease outbreak caused by VP, and other pathogenic factors was excluded.