The virulent genes are the key players in the ability of the bacterium to cause disease. The products of such genes that facilitate the successful colonization and survival of the bacterium in or cause damage to the host are pathogenicity determinants. This study aimed to investigate the prevalence of virulence factors (esp, agg, gelE, CylA) in E. faecalis isolated from diverse human clinical collected in Iraqi patient , as well as to assess their ability to form biofilm and to determine their haemolytic and gelatinase activities. Thirty-two isolates of bacteria Enterococcus faecalis were obtained, including 15 isolates (46.87%) of the urine, 6 isolates (18.75%) for each of the stool and uterine secretions, and 5 isolates (15.62%) of the wounds from various hospitals in Baghdad, including (Central Children's Hospital, Educational Laboratories, Ibn Al-Baladi Hospital).The isolates were confirmed to belong to the genus E.faecalis after performing morphological and biochemical microscopic examinations and for final diagnosis using the VITEC 2 system. The virulence genes viz. cylA, esp, gelE and agg were recognized in the E. faecalis, and the consequences appeared that the bacteria had eps gene in 32 isolates (100%). As for the agg gene, 32 isolates (100%) were carriers of this gene, which was responsible for these isolates' aptitude to form the biofilm. While for the gelE gene, 27 isolates (84.37%) of the isolates carried this gene, responsible for gelatinase activity whereas, the gene responsible for hemolysis cyl, there were 29 isolates (90.62%) of the total isolates. The presemce of genes in the isolates would be helpful to determine the colonization and survival of the bacterium in or causing damage to the host.