Background: The objective of this study was to determine the prevalence and antimicrobial susceptibility profiles of the clinical enterococci isolated in relation to biofilm formation and associated demographic characteristics, risk factors and disease outcome. Methods and materials: This cross-sectional study was conducted by utilizing Modified Congo Red agar to screen for biofilm producers among all clinical enterococci isolated from January 2017 until December 2017 in Hospital Sungai Buloh. Results: Of 180 clinical enterococci isolated, 19 repetitive clinical samples were excluded. The remaining 161 clinical enterococci isolated were tested and only 59 (36.6%) isolates were biofilm producing enterococci. The biofilm producing enterococci were predominantly isolated among the Enterococcus faecalis (66.1%) followed by Enterococcus faecium (28.8%). The isolates were equally distributed among urine (37.2%) and blood samples (37.2%), followed by tissue samples (23.7%). Surprisingly, antimicrobial susceptibility profile of the biofilm producing enterococci revealed that the isolates were sensitive to ampicillin (69.5%), high level gentamicin (86.4%), vancomycin (84.7%), and linezolid (94.9%) respectively. The biofilm producing enterococci were predominantly isolated from male (69.5%) patients with age less than 65 years old (55.9%). Mean age of patients with biofilm producing enterococcal infection was 57.3 years. More biofilm producing enterococci were isolated from patients with underlying diabetes mellitus (64.4%) relatively to patients with underlying chronic kidney disease (33.9%). Biofilm associated enterococcal infection was more common among patients with intravascular catheter (44.1%) and urinary catheterization (42.4%). There was a significant association between mortality and biofilm associated enterococcal infection (p-value = 0.002). Conclusion: In conclusion, the prevalence of biofilm producing enterococci necessitate a need for routine diagnostic method as it represents a third (36.6%) of all enterococcal infection especially among patients of male in gender and above 55 years of age. High index of suspicion for biofilm enterococcal infection is required among patients with underlying diabetes mellitus and presence of indwelling catheterization.