Summary Diabetic foot ulcers (DFUs) are a significant complication of Diabetes Mellitus, leading to reduced quality of life and even mortality in severe cases. Factors such as the immunocompromised state of the patient, hyperglycemia, and ischemia contribute to microbial colonization of DFUs, often resulting in severe complications like gangrene, osteomyelitis, and sepsis. Despite advancements in treatment modalities, the standard treatment still involves the use of parenteral broad-spectrum antibiotics. This dependency contributes to the emergence of antibiotic resistance. This prospective observational study aims to isolate and identify microorganisms from DFUs and determine their antibiotic susceptibility patterns. Swab samples collected from 15 diabetic patients undergo Gram staining, culture, and biomedical testing for microbial identification. This is followed by testing for antibiotic sensitivity to determine the pattern of antibiotic resistance in the isolated organisms. Results indicate Escherichia coli as the most common organism isolated. Notably, all Staphylococcus isolated were methicillin-resistant (MRSA), and 60% of the E. coli isolates exhibit Extended-spectrum beta-lactamase (ESBL) production. Resistance of Proteus mirabilis to Tetracycline and Polymyxin B and that of Enterococcus faecalis to Tetracycline is also noteworthy. In conclusion, this study shows the complexity of Diabetic foot ulcers and the increasing threat of antibiotic resistance in clinical practice.The findings indicate the importance of developing a more targeted and effective treatment strategy tailored to individual patients, there by reducing the risk of antimicrobial resistance, and enhancing patient outcomes in DFU management.