Background:  About 15% to 25% of people with diabetes will develop a foot ulcer. These wounds are often resistant to healing; therefore, people with diabetes are 20 times more likely to experience lower limb amputations than non-diabetic individuals. Aim of the study: To identify the causative organisms causing diabetic foot ulcers (DFUs) and to determine their antibiotic susceptibility. Methods: This is an observational cross-sectional study that included 100 different diabetic foot wound specimens collected from the patients attending at the vascular outpatient clinics of Cairo University Hospitals over the period from April 2022 to October 2022. Antibiotic susceptibility was identified by disc diffusion method and MIC. Results: The prevalence of Gram-negative isolates (75.4%) (89/118) was more than the Gram-positive (22%) (26/118). The most common isolated organisms were Klebsiella spp. (24%) (28/118 isolates), Proteus spp. (17.8%) (21/118 isolates), Pseudomonas spp. (16%) (19/118 isolates) and Staphylococcus aureus (13.5%) (16/118 isolates). Three Candida albicans isolates were recovered from the 118 isolates (2.6%). Multidrug-resistance (MDR) was detected in 67% (79/118 isolates), extensive drug resistant (XDR) was found in 24.5% (29/118 isolates). Extended-spectrum β-lactamase (ESBL) was found in 45% of the Gram-negative isolates (40/89 isolates), 22.5% of the Gram-negative isolates were carbapenem resistant Enterobacteriaceae (CRE) (20/89 isolates) and 46% of the Gram-positive isolates were methicillin-resistant Staphylococcus aureus (MRSA) (12/26 isolates). Colistin resistance was found in 6% (4/66) of the Gram-negative isolates by broth microdilution method. Conclusions: DFUs are mostly monomicrobial, more in type 2 DM. As per Wagner's classification, the prevalence of grade 3 ulcers is the highest.