In this study, biofilm formation and antibiotic resistance of Staphylococcus aureus strains isolated from Erzincan tulum cheeses were phenotypically and genotypically investigated. Samples collected from 100 different Erzincan tulum cheese were inoculated into Baird-Parker agar to isolate S. aureus. Identification of S. aureus was performed with Gram staining, hemolysis or coagulase tests. Determination of the nuc, mecA, vanA, and vanB genes in isolates was performed by Polymerase Chain Reaction (PCR). Congo red agar was used for the biofilm formation of S. aureus. Antibiotic resistance was determined by antibiotic discs of oxacillin (1 mu g), cefoxitin (30 mu g), vancomycin (30 mu g), amoxicillin-clavulanic acid (20 mu g), and penicillin (10 units). A total of 72 of 100 (72%) samples were positive for Staphylococcus spp. Of 72 samples, 61 (84.7%) were phenotypically and genotypically identified as S. aureus. Of 61 isolates, 37 (60.6%) formed a biofilm. Of 61 isolates, 49 were determined to resistant to antibiotics of oxacillin (methicillin) (9), cefoxitin (8), amoxicillin-clavulanic acid (4), and, penicillin (28). Vancomycin-resistance was not detected. Only the nuc and mecA genes were detected in 10 of 61 (16.3%) strains of S. aureus. In this study, the rate of S. aureus determined in Erzincan tulum cheeses was high. Considering the high rate of contamination and antibiotic resistance due to poor hygienic conditions, it was concluded that Erzincan tulum cheese, now a PDO cheese, should be considered to be great risk for public health.