The main objective of this study was to isolate and identify Staphylococcus aureus from retail raw red meat samples and evaluate their enterotoxin gene and antibiotic resistance profiles. A total of 452 retail raw meat samples, including beef (n = 200), sheep (n = 125), and lamb (n = 127) randomly purchased from various supermarkets and butchers in Ankara between July 2019 and November 2020, were tested for the prevalence of S. aureus. The S. aureus strain was identified using morphological and molecular (16S rRNA and nuc gene) methods. Moreover, nine Staphylococcal enterotoxin (SE) genes were screened using polymerase chain reaction. Antibiotic resistance of S. aureus was determined using the phenotypic disc diffusion method. The overall prevalence of S. aureus among screened samples was 21.23%. Additionally, 65.62% of S. aureus strains contained SE gene regions. The predominant SEs in the S. aureus strains were sea (50.79%), followed by sed (25.39%) and seb (23.80%). However, sec, see, seg, seh, sei, and sej genes were never detected. A substantial proportion (40-100%) of the isolates were found resistant to kanamycin, telithromycin, penicillin G, streptomycin, erythromycin, cloxacillin, ampicillin, pristinamycin, nalidixic acid, azithromycin, and ciprofloxacin. Multi-drug resistance (MDR) was observed in 96.87% of the S. aureus strains. These results show a low prevalence of S. aureus in raw red meat samples in Turkey. However, a high rate of SEA raises serious health concerns. Due to the high levels of MDR observed in this study, there is a need to strictly control antibiotic use in animals in Turkey.