Aim: The aim of this study was to determine the frequency of two important pathogenic microorganisms associated with endodontic infections, Enterococcus faecalis and Candida albicans, in root canal samples from patients with necrotic pulps or failed canal therapy by polymerase chain reaction method. Methodology: Microbial samples were obtained from 117 teeth with necrotic pulp tissues and 114 teeth with failed endodontic treatment. Results: E.faecalis were identified in 16% of the necrotic and 10% of the retreated root canal infections by PCR. C.albicans genome were identified in 20% and 11% of the necrotic and retreated root canal infections, respectively, by PCR. The frequencies of microbiota were not statistically different between necrotic and retreatment groups (p > 0.05, chi squared test). Conclusions: PCR analysis of teeth with periapical lesions revealed that E.faecalis was found in fewer patients than in previous studies. The C.albicans prevelance was consistent with previous reports. No statistical difference was found between primary and secondary root canal infections for C.albicans or E.faecalis.