Background: Staphylococcus aureus is one of the main human pathogens which should be taken into account as one of the main hospital infections due to its colonization in the nose of carrier personnel.The present study aims to determine the frequency of resistance to mupirocin in the hospital carriers of Staphylococcus aureus, and to investigate the antimicrobial effect of Zinc Oxide nanoparticles on the growth of resistant strains. Methods: In this cross-sectional study, samples were taken from the anterior nasal part of 150 hospital staff in Gorgan, using a sterile swab. Being cultured on Mannitol salt agar, the suspected colonies were identified through Gram staining, as well as catalase, coagulase and DNase tests. Resistance of the strains to mupirocin was tested using microdilution Broth test. Besides, the antibacterial effect of Zinc Oxide nanoparticles(ZnONPs) on the mupirocin-resistant strains was also investigated using agar well diffusion method. Results: In this study, the frequency of Staphylococcus aureus was 32%, out of which 6.2% showed high resistance to mupirocin and 29.2% showed low resistance to this antibiotic. The results of this study showed that Zinc Oxide nanoparticles have the most inhibitory effect on the growth of mupirocin-resistant Staphylococcus aureus in the density of 400 mg/ml. Conclusion:The Zinc Oxide nanoparticle used in this study had a high antimicrobial activity against all the drug-resistant Staphylococcus aureus strains, and their bactericidal properties also increased by raising the density of the nanoparticle.