OBJECTIVE This study aimed to examine the reliability of the 512-Hz Rinne tuning fork test to detect conductive hearing losses. The effects of tester experience, the use of masking, and the interpretation of equivocal (+/-) Rinne results on test reliability also were examined. METHODS Retrospective. METHODS Private otology practice. METHODS 1,000 adult patients (2,000 ears) seen for their initial otologic evaluation. METHODS Diagnostic. METHODS Sensitivity of the 512-Hz Rinne tuning fork test was assessed by comparing tuning fork results with the pure-tone average air-bone gap. RESULTS Results showed the 512-Hz Rinne tuning fork test could be very effective at detecting conductive hearing losses when performed by an experienced tester and when masking was used. Sensitivity was lower when masking was not used and lowest when the Rinne was performed by a less-experienced tester. Sensitivity for all groups was improved by interpreting equivocal results as indicating a conductive loss. CONCLUSIONS Despite reports of poor reliability, the 512-Hz Rinne tuning fork test can be an important tool in an otology practice for the detection of conductive hearing losses and for confirming audiometric findings. In primary care settings, the Rinne would be most effective as part of a screening program for conductive hearing losses, but not as the sole indicator for referral.