Staphylococcus aureus (S. aureus) is among the most virulent bacterium, causing a wide spectrum of diseases, one of which is sinusitis. Sinusitis is defined as inflammation of the paranasal sinuses and can occur with an S. aureus infection. Since traditional diagnosis of sinusitis, such as bacterial culture, can be very time-consuming, there is an urgent need to implement a rapid and reliable method to detect S. aureus sinusitis. In this study, 55 sinusitis samples were provided from operating rooms in two ways: puncture and endoscopic surgery. The application of Polymerase Chain Reaction (PCR) in the detection of S. aureus in sinusitis samples was evaluated. The PCR test was performed with the S. aureus nuc gene, and produced a 279 bp DNA band, using 1.5% agarose gel electrophoresis. Finally, according to PCR analysis, 18 out of the 55 studied specimens (32.7%) from patients with sinusitis were positive for S. aureus. These results indicate that the use of the PCR method is a highly specific, sensitive, and reliable tool for the rapid detection of S. aureus-derived sinusitis. The results of this report will aid physicians in the appropriation and prescribing of antibiotics.