Objective:To identify the cause of a suspected foodborne disease caused by bacterial food poisoning using multiple detection methods.Methods:At 9:35 a.m. on August 9, 2022, a suspected foodborne disease incident was handled by the Hefei Center for Disease Control and Prevention. The preliminary laboratory test results showed that the foodborne disease was caused by Staphylococcus aureus infection. Sixteen strains of Staphylococcus aureus were isolated, cultured, and identified using real-time fluorescent polymerase chain reaction. At the same time, the 16 strains of Staphylococcus aureus isolated from different sources were analyzed by pulsed-field gel electrophoresis to determine the correlation between strains. Results:The distribution of 16 strains of Staphylococcus aureus enterotoxins isolated was that the anal swab and chopping board (inner) smear samples from a salesperson surnamed Li showed SEB type, and the remaining 14 strains all carried type A, C, and E enterotoxin genes simultaneously. Pulsed-field gel electrophoresis typing results showed that there were two types of strains; except the anal swab and chopping board (inner) smear samples from the salesperson surnamed Li, the other 14 strains were all of the same type, with a similarity of 100%. The similarity between the anal swab and the chopping board (inner) smear samples from the salesperson surnamed Li was 80.65%, and the similarity with another type was 59.26%. This result was consistent with the detection results of Staphylococcus aureus enterotoxins. Conclusion:The foodborne disease in this case was caused by mixed contamination of two or three different sources of Staphylococcus aureus with enterotoxins.