Objective Laboratorial detection were performed on a case of a food poisoning. Meanwhile, pulse field gel electrophoresis (PFGE) and antimicrobial susceptibility testing were carried out for pathogens obtained. Methods Suspected milk and ice cream, vomitus and feces specimens were tested for Salmonella, Shigella, diarrheogenic Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, and Norovirus in accordance with the method of national food safety standard and others. Analysis of enterotoxins, PFGE and antimicrobial susceptibility testing of Staphylococcus aureus isolates were performed. Results Totally, twelve strains of Staphylococcus aureus from milk and ice cream specimens, four strains from the vomitus and the fecal specimens of the patient were cultured. All these strains were found to be able to produce type A, C, and E enterotoxins. Further molecular subtyping by PFGE analysis showed that these strains produced indistinguishable pulsotypes. Moreover, these strains were resistant to penicillin and erythromycin. Conclusion This food poisoning outbreak was associated with the milk contaminated with Staphylococcus aureus stains and produced large amounts of enterotoxin, and the ice cream processing centers used the contaminated milk.It is recommended that the supervision department should strengthen the supervision of the food producers especially in rural areas and milk supply stations in order to prevent the outbreaks of similar incident.