Background: The enterotoxigenic Staphylococcus aureus is considered as one of the most important cause of food poisoning that manifests with gastroenteritis, diarrhea, and vomiting. Its complications usually occur when bacterial virulence genes are produced. The most important virulence factors are cell-associated components, exoenzymes, exotoxins, enterotoxins, and enterotoxin-like toxins.Objectives: The present study aimed to study the presence of S. aureus and its virulence factors in chicken nuggets in Iran.Materials and Methods: Totally, 420 chicken nuggets from five brands were collected from Isfahan and Chaharmahal-va-Bakhtiari provinces, Iran. Samples were cultured and the positive results were studied using ELISA and PCR for detection of classical staphylococcal enterotoxins and sea-sej virulence genes, respectively.Results: Results showed that 27 (6.42%) of 420 samples were contaminated with S. aureus with bacteria concentration between 6.1×103 to 8.4×101/mL. Totally, 33.33% of isolates produced SEA, 4.16% SEB, 12.50% SEC, 8.33% SED, 12.50% SEA+SEC, and 12.50% SEA+SED. The most commonly detected genes were sea (25%), sea+seg (8.33%), sec (12.50%), sea+sed (12.50%), and sea+sec+sej (12.50%).Conclusions: S. aureus can easily contaminate the chicken nugget and this contamination is usually associated with significant presences of virulence genes. Consumption of these nuggets certainly is associated with gastrointestinal diseases. Therefore, some food safety and quality standards should be applied and performed in most of the Iranian food units to control growth of S. aureus and its virulence factors.