Staphylococcus aureus is an important cause of gangrenous mastitis (GM) in dairy farming and causes significant economic losses due to toxins such as alpha toxin, beta toxin, fibronectin binding protein, and leucocidins. Alpha toxin is defined as the primary virulence factor that initiates necrosis in udders. In this study, the virulence gene profile of S. aureus strains isolated from milk with GM was compared with isolates obtained from non-gangrenous mastitis cases. According to the data obtained, virulence gene profiles of mastitis isolates were compared with GM isolates and virulence gene similarity between mastitis strains and GM strains was investigated. Nine S. aureus strains were isolated from 85 milk samples and analysed for virulence factors causing GM. By PCR genomic analyses, 9 (10.5%) S. aureus strains were isolated. The isolates were positive for alpha toxin (44.4%), beta toxin (44.4%), leukocidin F (55.5%), fibronectin binding protein A (55.5%) by PCR. The strains carrying virulence genes, especially alpha toxin, were found to be associated with GM. Antibiotic resistance profiles were determined and found to be significantly resistant to various antibiotics. In the medium test for Hla production, Brain Heart Broth and Tryptic Soy Broth were found to be suitable for Hla production. PCR analyses showed that strains carrying more than one virulence gene, especially Hla, were obtained from strains with GM and statistical analyses showed that the isolates with GM did not resemble strains with mastitis in terms of virulence gene profile.