Background: It has been found that 80% of microbial infections are associated with biofilm formation. For example, urinary tract infections, middle-ear infections, formation of dental plaque, endocarditis and cystic fibrosis, biofilms are also found to be associated with indwelling medical devices such as contact lenses, central venous catheters and mechanical heart valves. Objectives: The aim of this study was to evaluate the ability of the isolated organisms causing CAUTIs to form biofilm, as well as the effect of EDTA, Kojic acid and their combination on inhibiting their biofilm formation. Methodology: The bacterial isolates were tested for their ability to form biofilm by Inoculum preparation, Washing, Fixation, Staining and Interpretation of the results by detecting their optical density (OD) of each well stained with crystal violet and measured at 620 nm using a Micro titre plate reader (Tecan. Infinit F50). Results: In this study the most frequently biofilm forming species was Pseudomonas 5(21.7%), while the least biofilm forming isolates were Enterococcus fecalis 2(8%). EDTA was the most powerful biofilm inhibitor as it inhibited biofilm by 78.6% with mean optical density reading 0.22 ± 0.15, EDTA and Kojic acid combination had nearly close results on biofilm as it decreased biofilm by 72.8% with mean optical density reading 0.28 ± 0.17, while Kojic acid decreased biofilm formation by 51.4% only with mean optical density reading 0.5 ± 0.31. Conclusion: EDTA was the most powerful biofilm inhibitor, EDTA and Kojic acid combination had nearly close results on biofilm to EDTA, while Kojic acid had the least effect on biofilm inhibition.