Diabetes mellitus is a major health concern, and foot ulcers are a serious complication. Pseudomonas aeruginosa is a common pathogen in diabetic foot infections, which can delay wound healing and increase morbidity. This study compares the diagnostic performance of conventional microbiological methods and molecular techniques (PCR) for identifying P. aeruginosa in diabetic foot ulcers and evaluates its antimicrobial susceptibility. In a cross-sectional study of 134 patients, samples were collected and analyzed using conventional culture methods and PCR. PCR was more efficient, detecting 44% of cases versus 14% by culture. Co-trimoxazole was the most resistant drug, while amikacin was the most sensitive. The study highlights the importance of molecular techniques for accurate, fast pathogen identification, crucial for appropriate treatment and improved clinical outcomes.