Diabetic foot ulcer (DFU) is one of the most common complications of diabetes mellitus caused by uncontrolled hyperglycemia and typically begins with minor cuts that lead to skin ulceration. Untreated ulcers may develop an infection over time and may persist despite rigorous treatment strategies, and could be due to the presence of biofilms. A biofilm is a layer of bacteria attached strongly to a surface and encased in a tough and slimy substance called extracellular polymeric substance (EPS). This highly virulent bacterial layer contributes to antimicrobial resistance and wound chronicity among DFU patients. This project aims to isolate and identify the bacteria in wounds of DFU patients in Orthopedic Clinic in Serdang Hospital and to investigate their antibiotic sensitivity and their biofilm-forming capabilities. Majority of the patients were Malay of 56-65 years, presenting with Wagner Grade 2 ulcer and had the ulcer for over 4 weeks. Bacteria were isolated from the wound samples and had a high prevalence of Gram-positive bacteria (60.76%). Majority of the bacterial species were able to form biofilms and were most sensitive to imipenem and most resistant to penicillin G. In conclusion, we found that DFU is persistent in elderly male diabetic patients, most of whom only began seeking medical attention for their wound one month from its onset. Bacterial isolation performed on the samples showed that DFU wounds tend to be polymicrobial. The isolates also demonstrate increasing resistance against commonly used antibiotics for treating DFU, which can be worrisome for both clinicians and patients.