Background: Intracanal microorganisms are the main cause of radicular lesions and, therefore, should be removed from the root canal system. Enterococcus faecalis is a resistant bacterium isolated from 30%–90% of endodontically-treated teeth, commonly chosen for in vitro studies on endodontic treatment methods. Different methods have been suggested for removing this bacterium from the root canal system, while none have been approved as the gold standard method. This study aimed to compare the effectiveness of Edge Taper Platinum and XP-endo® shaper rotary files in reducing the number of E. faecalis in the root canal system. Methods: In this ex vivo study, 58 single-rooted single-canal teeth were immersed in formalin for 1 hour, stored in 0.9% saline, and randomly divided into positive control (n=4), negative control (n=4), platinum (n=25), and shaper (n=25). E. faecalis was injected into the root canals of all groups, except for the negative control group. The teeth were then incubated in a CO2 incubator for 24 hours. Primary samples were collected, and the root canals were instrumented using the XP-endo® shaper and Edge Taper Platinum. Secondary samples were collected and cultured, and the number of colonies was counted after 24 hours of incubation. The data were analyzed using the Kruskal-Wallis and Mann-Whitney tests at a P<0.05. Results: A significant difference was observed for the reduction of the E. faecalis colony count in 87.79%, 97.83%, and 14.11% of the XP-endo® shaper (87.79%), Edge Taper Platinum (97.83%), and positive control (14.11%) groups (P<0.05). Conclusion: Both XP-endo® shaper and Edge Taper Platinum files with equal apical sizes effectively reduced the number of E. faecalis colonies in the root canal system. Edge Taper Platinum with a greater taper was more effective.