The incidence of Methicillin-resistant Staphylococcus aureus (MRSA) infection poses significant challenges to epidemiology, infection control, and therapeutic management, particularly within intensive care units (ICUs). This study aimed to monitor MRSA infection rates among patients in various ICUs at Luxor International Hospital (LIH), one of the largest healthcare facilities in Upper Egypt. Mannitol Salt Agar (MSA), Cefoxitin disk (mecA-mediated Oxacillin resistance), Oxacillin Resistance Screening Agar Base (ORSAB), and VITEK2 COMPACT system were used for the isolation and identification of the bacterial isolates from various biological specimens. 69.6% of the staphylococcal isolates obtained from ICUs were identified as MRSA strains. Infection rates were 69.8% and 31.2% in males and females, respectively. The isolation rates of MRSA from the collected samples were as follows: blood (77.9%), sputum (11.3%), pus (9.0%), and urine (1.8%). MRSA infection was very prevalent in Moderate intensive care unit (MCU) and Neurological intensive care unit (NECU) compared to other ICUs. Antimicrobial susceptibility testing shows that the isolated MRSA strains are highly sensitive to Tigecycline, Linezolid, Vancomycin, Teicoplanin, and Rifampicin while they are highly resistant to Cefoxitin and Penicillin. Collectively, this study suggests that the prevalence of MRSA infection is high among ICUs patients, particularly MCU and NEICU patients. Findings from this study have important implications for future epidemiology, and infection control studies to develop an effective protocol to manage MRSA infection within settings.