Tehran University of Medical Sciences Iranian Journal of Microbiology 2008-3289 17 2 2025 04 05 229 238 Rania Al-Groom Department of Medical Laboratory Science, Faculty of Allied Medical Sciences, Zarqa University, Zarqa, Jordan; Department of Allied Medical Sciences, Zarqa University College, Al-Balqa Applied University, Zarqa, Jordan Ghina Al-Saraireh Department of Medical Laboratory Science, Faculty of Allied Medical Sciences, Zarqa University, Zarqa, Jordan Sultan Ayesh Mohammed Saghir Department of Medical Analysis, Princess Aisha Bint Al-Hussein College of Nursing and Medical Sciences, Al-Hussein Bin Talal University, Ma`an, Jordan Mohd Sajjad Ahmad Khan Department of Basic Sciences, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia Areej Almanaseer Department of Medical Laboratory Science, Faculty of Allied Medical Sciences, Zarqa University, Zarqa, Jordan Laila Alswalha Department of Allied Medical Sciences, Zarqa University College, Al-Balqa Applied University, Zarqa, Jordan Wesal Alraei Department of Diet Therapy Technology and Dietetics, Faculty of Allied Medical Sciences, Zarqa University, Al-Zarqa, Jordan Dalia Abu Al-Haijaa Department of Diet Therapy Technology and Dietetics, Faculty of Allied Medical Sciences, Zarqa University, Al-Zarqa, Jordan Maha Hdaib Department of Allied Medical Sciences, Zarqa University College, Al-Balqa Applied University, Zarqa, Jordan Anas Da'meh Department of Medical Laboratory Science, Faculty of Allied Medical Sciences, Zarqa University, Zarqa, Jordan Shereen Burjaq Department of Medical Laboratory Sciences, Faculty of Science, Al-Balqa Applied University, Salt, Jordan Omar Al-Dmour Department of Allied Medical Sciences, Zarqa University college, Al-Balqa Applied University, Salt, Jordan Fuad Alhawarat Department of Applied Medical Sciences, Al Hussein Bin Abdullah II Academy for Civil Protection, Al-Balqa Applied University, Salt, Jordan 2024 11 24 2025 03 30 Background and Objectives: Staphylococcus aureus (S. aureus) is one of the most important pathogens, responsible for a range of infections. This study aimed to assess resistance patterns in S. aureus isolates obtained from certain private-sector laboratories against commonly used antimicrobial agents. Materials and Methods: The process involved collecting various samples from several private laboratories and then identifying S. aureus isolates using biochemical characterization. The antibiotic susceptibility of these isolates was determined by disc diffusion method . Furthermore, Rt-PCR was employed to identify two genes namely the methicillin/oxacillin resistance genes (mecA), and (SCCmec). Results: The findings of the current study exhibited that females constituted a larger proportion of the participants (59.1%) compared to males (40.9%), with a mean participant age of 40.82 years. Gram-positive bacteria were more prevalent (71.3%) than Gram-negative bacteria (18.3%), with S. aureus being the most frequent isolate (60.9%). Urine samples represented the highest collected sample type (47.8%). Out of the 115 bacterial isolates, 85.2% exhibited multidrug resistance to antibiotics such as cefazolin, gentamicin, vancomycin, and ceftazidime. Clindamycin was the most effective antibiotic, with a sensitivity rate of 62.9%, followed by teicoplanin and meropenem, each with a sensitivity rate of 52.9%. Methicillin-resistant Staphylococcus aureus (MRSA) strains were susceptabile to vancomycin and teicoplanin. The methicillin/oxacillin resistant isolates showed significant association with mecA and SCCA genes. Conclusion: This study highlighted the multi-drug resistance in S. aureus isolates, stressing the need for stringent antibiotic stewardship, continuous surveillance, and further research into alternative treatments, including novel antibiotics and combination therapy, to combat resistant strains. https://ijm.tums.ac.ir/index.php/ijm/article/view/5153 https://ijm.tums.ac.ir/index.php/ijm/article/download/5153/1761