OBJECTIVE: To assess the efficacy of a standard cleaning and sterilization protocol employed during reuse of cardiac electrophysiology catheters on the infectivity of duck hepatitis B virus (DHBV; a surrogate for human hepatitis B virus), bovine viral diarrhea virus (BVDV; a surrogate for human hepatitis C virus), and human coxsackie type B3 virus (CB3). SETTING: Public health virology laboratory. METHODS: Studies were performed on the distal, electrode-containing segments of 120 electrophysiology catheters previously used in up to 10 clinical procedures. Catheter segments were immersed for 1 hour in blood infected with high titers of DHBV, BVDV, or CB3. After air drying for 2 hours, subgroups of 8 catheters were subjected to no treatment, washing in general-purpose detergent, washing in enzyme cleaner, sterilization in ethylene oxide, or the full protocol of sequential detergent-enzyme cleaner-ethylene oxide exposure. Presence of residual virus was assessed by nucleic acid detection and infectivity studies. RESULTS: DHBV nucleic acid was detected on catheters after individual steps and the full protocol, whereas BVDV and CB3 nucleic acids were detected after individual steps but not the full protocol. These findings were associated with the presence of infectious DHBV and CB3, but not BVDV, on catheters after washing in detergent or enzyme cleaner. However, ethylene oxide alone or the full protocol reduced infectivity of all three viruses to undetectable levels. CONCLUSION: These experimental studies provide strong evidence that appropriate cleaning and sterilization of reused electrophysiology catheters inactivates blood-borne viruses such as hepatitis B and C and coxsackie type B3.