Background: Antibiotic resistance is a critical global public health crisis, particularly regarding nosocomial infections acquired within healthcare settings. These infections are frequently caused by multidrug-resistant (MDR) bacteria, leading to increased morbidity, mortality, and healthcare costs. The World Health Organization (WHO) has highlighted the urgent need for comprehensive surveillance of antibiotic resistance patterns to inform treatment strategies and infection control measures.Objective:This study aims to analyze antibiotic resistance patterns among pathogenic bacterial isolates from nosocomial infections in Menoufia, Qalubiya, and Behira governorates in Egypt, examining their susceptibility to commonly used antibiotics.Materials and Methods:A cross-sectional analysis was conducted on 100 clinical samples collected from patients diagnosed with nosocomial infections in the three governorates. Samples underwent bacterial isolation, morphological identification, and antibiotic susceptibility testing using the Kirby-Bauer disk diffusion method. The isolates' morphological, physiological, and biochemical characteristics were assessed, followed by statistical analysis using Chi-square and ANOVA tests.Results:The analysis revealed a concerning prevalence of MDR bacteria, including Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa, all exhibiting 100% resistance to penicillin, methicillin, erythromycin, and tetracycline. Resistance distribution showed no significant differences among the three governorates. Most isolates were identified as Gram-negative bacilli, with only Staphylococcus aureus and Enterococcus faecalis as Gram-positive. Physiologically, the majority thrived at 37°C and neutral pH. Biochemical tests confirmed the identities of the isolates, showing uniform characteristics across strains.Conclusion:This study highlights the challenges posed by high MDR bacteria prevalence in nosocomial infections, emphasizing the need for comprehensive infection control strategies and antibiotic stewardship programs to combat antibiotic resistance effectively