Abstract : A total of 10 bacterial isolates were obtained from 100 specimens collected from diabetic foot ulcers from patients of different age groups and gender attended to the consulting clinic at Baghdad teaching hospital in Baghdad city during the period from September 2019 to January 2020. Isolates were initially diagnosed by note morphological and cultivar traits on Blood agar, Enterococcus agar, Bile esculin azide agar and Nutrient agar. These colonies of E. faecalis appears as non-hemolytic (gamma-hemolytic) colonies on blood agar with small, white circular colonies on Nutrient agar. While microscopic appearance of E. faecalis was Gram positive cocci spherical in shape with a diameter of less than 1 micron arranged in single, double or short strings. Bacterial isolates suspected to E. faecalis were identified according to their biochemical properties and these results were confirmed by Vitek-2 system. Among the ten E. faecalis isolates tested for biofilm production, 3 (30%) E. faecalis isolates were countering strong biofilm productions, one (10%) isolates moderate and six (60%) isolates weak biofilm produce as shown in figure (4-6). The optical density (OD) was measured at 570 nm. Results of molecular study documented the amplification of (asa1) gene that encoded aggregation substance in E. faecalis through real time PCR Assay showed there were only one (10%) isolates that negative for asa1 gen and nine (90%) isolates were positive finding. Also the results of cycling GelE gene showed that all isolates 10 (100%) have GelE gene. While the cycling CylA gene were revealed only 2 (20%) of isolates were negative for CylA gen and 8 (80%) isolates positive for. In addition, the results of cycling Esp gene were showed only 2 (20%) of E. faecalis isolates were negative for Esp gen and the positive isolates were 8 (80%) and results of cycling HylA gene presented that all isolates negative for HylA gen 10 (100%).