OBJECTIVE To optimize the cleaning process of reusable rigid endoscopes, evaluate the cleaning effect by using ATP biology fluorescence method and microorganism cultivation, and explore the impact of different cleaning process on the cleaning of reusable rigid endoscopes so as to improve the cleaning quality. METHODS 600 sets of contaminated rigid endoscopes collected from Nov. 2021 to Nov. 2022, are chosen as the research object. They were randomly divided into two experimental group and the control group according to the cleaning process. For the control group, the contaminated instruments followed the procedure: water washing→check→disassemble→ enzyme soaking→high-pressure spray→ultrasonic cleaning→pure water rinse→thermal disinfection(90℃-5min)→lubricate→dry；For the experiment group, the cleaning process is washing→check→disassemble→high-pressure spray→Automatic machine wash(including enzyme wash、alkaline-wash、ultrasonic-cleaning、lubricate、thermal disinfection and dry ). All the instruments washed and disinfected were examined by means of ATP biology fluorescence test and microorganism cultivation, and the difference value of visual inspection and ATP biology fluorescence test were calculated and compared. RESULTS The qualified rate of processed instruments of the two groups(control group、experiment group) are 93.3% and 97.3% by microorganism cultivation, 87.7% and 94.0% by ATP bioluminescence essay(P<0.05). CONCLUSION The cleaning process utilized hand washing together with professional rigid endoscopes cleaning frame can significantly improve the cleaning quality of the reusable rigid endoscopes , and could achieve better cleaning effect.