Pseudomonas aeruginosa is a critical bacterial pathogen that plays a major role in hospital-acquired infections, particularly among patients with weakened immune systems, chronic illnesses, or severe burns. The treatment of these infections has become challenging due to the bacterium’s broad-spectrum antibiotic resistance. This study aimed to isolate and identify carbapenem-resistant P. aeruginosa strains from clinical samples. A total of 126 P. aeruginosa isolates were collected from various departments of Imam Reza Hospital. The isolates were identified using biochemical methods. Antibiotic susceptibility was evaluated using ceftazidime, ciprofloxacin, piperacillin-tazobactam, amikacin, gentamicin, and cefepime, while the minimum inhibitory concentration for imipenem was determined using the E-test. The prevalence of bla_GIM and bla_SHV genes was assessed through PCR. The results showed that 41.26% of the isolates were obtained from urine cultures, followed by body secretions (35.15%) and tracheal samples (10.31%). The highest frequency of isolates (26%) was recorded in the intensive care unit. Among the 126 isolates, resistance was most prevalent against ciprofloxacin (73%) and least against amikacin (61.2%). Additionally, 69% of isolates were resistant to both imipenem and ceftazidime, 68% to cefepime, and 65% to gentamicin and piperacillin-tazobactam. Carbapenem-resistant strains exhibited MIC values exceeding 32 µg/ml. A total of 81 multidrug-resistant were identified. While the bla_GIM gene was not detected in any isolate, 71% of the samples harbored the bla_SHV gene. The high prevalence of antibiotic resistance observed in this study highlights the significant impact of unregulated and excessive antibiotic use. This growing resistance substantially complicates the effective treatment of P. aeruginosa infections.