Objective To evaluate the effect of axial-flow cabinet mechanical ventilation, exhaust fan mechanical ventilation and natural ventilation on cleaning and disinfecting aerosol in digestive endoscope cleaning and disinfection room. Methods The experiment was divided into three groups: Axial flow cabinet mechanical ventilation (group A), exhaust fan mechanical ventilation (group B) and natural ventilation (group C). Air in digestive endoscope cleaning and disinfection rooms in two wards of a hospital under three ventilation modes were collected: before and after 2 hours of ventilation in the static environment (the cleaning and disinfection work was stopped for 12 hours), 0 hour in the initial state of the dynamic environment (the cleaning and disinfection work was in progress) and 1, 2, 4, 6 hours of continuous ventilation, contents of particulate matter, microorganisms, glutaraldehyde in air aerosol were detected, effect of three ventilation modes on purifying the aerosol in digestive endoscope cleaning and disinfection room were compared. Results In static environment, contents of particulate matter, microorga-nism and glutaraldehyde in the cleaning and disinfection endoscope room showed a downward trend after 2 hours ventilation, differences between group A and B before and after 2 hours of ventilation were statistically significant (P < 0.05). At 0 hour of initial working state of cleaning and disinfection room, contents of particulate matter, microorganism and glutaraldehyde in air among three groups were not significantly different (all P>0.05). After continuous ventilation for 1, 2, 4 and 6 hours, contents of particulate matter, microorganism and glutaraldehyde in air among all groups and between any two groups were significantly different (all P < 0.01), group A was lower than group B and C, and group B was lower than group C. In group A, contents of particulate matter, microorganism and glutaraldehyde in dynamic environment was low and the fluctuation range was small, and content of particulate matter reached the specified range of class 1 million clean room. Contents of glutaraldehyde in group B and C exceeded the maximum safe exposure limit of 0.175 mg/m³ at 1, 2 and 4 hours of continuous ventilation, content of gluta- raldehyde in group C still exceeded the maximum safe exposure limit during the lunch break of 4-6 hours. Conclusion Among three ventilation modes, axial flow cabinet mechanical ventilation mode can most effectively purify the aerosol in air of digestive endoscope cleaning and disinfection room, and it is recommended to introduce axial flow cabinet ventilation mode to ensure the occupational safety of staff.