OBJECTIVESTo investigate the impact of different pretreatment strategies and cleaning methods on the cleaning efficacy of electrosurgical instruments, with the aim of proposing an optimal protocol.METHODSIn total, 573 electrosurgical instruments were collected from three large-scale hospitals within 2 h of use between December 2023 and July 2024. The instruments were categorized into six groups: medical alkaline cleaner, multi-enzyme cleaner and instant foam-type multi-enzyme humectant followed by manual cleaning; and medical alkaline cleaner, multi-enzyme cleaner and instant foam-type multi-enzyme humectant followed by mechanical cleaning with a vacuum boiling washer. Cleaning quality was assessed through visual estimation, residual protein detection, and adenosine triphosphate (ATP) biological fluorescence detection.RESULTSVisual assessment indicated that different pretreatment and cleaning methods did not affect cleaning quality significantly (P>0.05). Residual protein detection analysis revealed that mechanical cleaning outperformed manual cleaning [adjusted odds ratio (AOR) 2.20, 95% confidence interval (CI) 1.18-4.11; P=0.013], with both medical multi-enzyme cleaners (AOR 3.72, 95% CI 1.11-12.44; P=0.033) and instant foam-type multi-enzyme humectants (AOR 3.27, 95% CI 1.07-10.03; P=0.038) combined with mechanical cleaning demonstrating superior performance. ATP detection confirmed that mechanical cleaning was more effective than manual cleaning (AOR 5.10, 95% CI 2.00-12.95; P<0.001), particularly when using a medical alkaline cleaner (AOR 6.79, 95% CI1.36-33.83; P=0.019) or a multi-enzyme cleaner (AOR 6.77, 95% CI 1.43-31.95; P=0.016) before mechanical cleaning.CONCLUSIONMechanical cleaning emerged as the critical factor for improving the cleaning efficacy of electrosurgical instruments, and no significant differences were observed between the three pretreatment strategies: medical alkaline cleaner, medical multi-enzyme cleaner, and instant foam-type multi-enzyme humectant. Furthermore, even electrosurgical instruments with complex structures maintain adequate cleanliness after standardized pretreatment and cleaning protocols.