Background and aims: Staphylococcus aureus is known as one of the most important foodborne pathogens worldwide. The aim of this study was to investigate the prevalence based on the biotype of S. aureus bacteria extracted from hospital foods. Methods: This descriptive cross-sectional study was conducted in a hospital catering in the south of Tehran city, in the three months of summer 2021. On each sampling occasion, based on the weekly cooking menu, a sample of rice, a sample of kebab, and two samples of cutlet were taken under standard conditions from the food of patients and staff. Results: In total, out of 411 food samples, 24 Isolates out of 127 kebab samples (18. 89%) and 16 Isolates out of 157 cutlet samples (10. 19%) were contaminated. There was no contamination in the rice sample. Also, there was a significant relationship between food cooking temperature and prevalence of S. aureus. In the comparison of the two contaminated samples, the highest prevalence of S. aureus was related to kebab (with cooking temperature of 160°, C) and the lowest prevalence was related to cutlet (with 180°, C). The prevalence rate of S. aureus in the kitchen surfaces of the hospital was 20%. The identified biotypes, in order from the highest to the lowest, include 15 (37. 5%) bovine, 13 (32. 5%) ovine, six (15%) poultry, four human (10%), two (5%) human β, ٭,biotypes. Conclusion: Improper cooking, serving, and transfer of cooked or processed foods, and poor consideration of hygiene factors play an important role in the spread of S. aureus bacteria.