The purpose of this study was to determine the time required for anaerobic bacteria in natural human saliva to contaminate root-filled teeth. Thirty-two single-rooted teeth were cleaned, shaped, filled, and exposed to human saliva for 120 days. Teeth that had not leaked were subjected to polymerise chain reaction examination. Sixty-six percent of the experimental group were totally contaminated. A PCR examination revealed there was no contamination in the apical 3 mm of leakage-free teeth