Enterococci are responsible for serious diseases such as urinary tract infections (cystitis, urethritis, pyelonephritis and prostatis), bacteremia, intra- abdominal, pelvic and soft tissue infections. Infection on burn surfaces were also commonly caused by Enterococcus. The ability of enterococci to cause such diseases is due to acquisition of certain virulence factors such as hemolysin, gelatinase, Biofilm formation and enterococcus surface protein (esp). This study has been conducted to investigate the occurrence of virulence factors between Enterococcus isolated from nosocomial infections and from stool samples (commensal isolates).Both clinical and stool specimens were collected and isolates were identified by microscopic, culture and standard biochemical tests. Virulence factors were examined by phenotypic tests and esp gene were detected by PCR method. There is no significant difference in the virulence factors like hemolysin and gelatinase production between clinical and commensal isolates. Biofilm production and presence of esp gene is very high among clinical isolates. Study of virulence factors associated with invasiveness and disease producing capacity of nosocomial pathogen like multidrug resistant Enterococcus will be very useful in treating them.