Background: Enterococcus bacteria are normal flora found in the gut of different organisms, this bacterium has the potential to become an opportunistic pathogen and harm the host's extra intestinal sites. Dental infections also associated with Enterococci. Objective: In order to shed light on multidrug resistance E. faecalis in root canals infections and also this research was carried out to investigate the percentage of asa-1, cyl genes A in the E. faecalis isolates from root canal samples by using polymerase chain reaction technique (PCR). Materials and Methods: During the period of November 2021 to January 2022, root canal samples were collected from (40) patients, (9) men and (31) women, and then were cultured on Pfizer specific Enterococcus media, a selective medium for the isolation and growth of Enterococcus spp, after that antibiotic sensitivity test was done on all E. faecalis isolated. And then molecular detection was done to detect the E. faecalis, asa-1 and Cyl A genes. Results: The results showed percentage of E. faecalis isolates from root canal was 55% (22 isolates) and all isolates were positive for E. faecalis gene, also 100% of isolates were positive for Cyl-A gene and 86.36% of isolates were positive asa-1 gene. The antibiotics sensitivity result showed that 100% of E. faecalis isolates were sensitive to Imipenem and Linezolid, whereas 100% of E. faecalis isolates were resistant to Tetracycline and Trimethoprim. Conclusion: Most of isolates were multidrug resistant and also almost all the isolates had asa-1 and Cyl-A genes together.