Objective To compare the cleaning quality and cleaning time of cleaning gastroscope with different cleaning processes by ATP bio-fluorescence method. Methods Sixty post-use gastroscopes with the same model (GIF-H290), production batch and input time were selected, and were randomly assigned to the experimental group (n=30) and the control group (n=30) with sealed envelopes method. Gastroscopes in the control group were used "scraping the mirror body - brushing the gastroscope tube - full line perfusion 2min - rinsing" process, while that in the experimental group were used "full line perfusion for 2min - scrubbing at the same time in the whole line perfusion Mirror body - brushing the gastroscope tube – rinsing” process. After completion, the ATP biofluorescence method was used to detect the ATP value of the cleaned gastroscopic surface and the gastroscopic tube, and the time taken for cleaning was also recorded. Among them, the scrubbing mirror body, the whole pipeline perfusion, the pipeline brushing and timing are completed by the A member, and the rinsing and detection are completed by the B member. The blind method is adopted for the B members (because the cleaning process can be judged by observing whether the whole body is perfused while observing the mirror body, and thus the grouping situation is known, the blind method is not adopted for the A member). The quality of the gastroscope cleaning and the cleaning time were compared between the two groups. Results There was no significant difference in the ATP value between the gastroscope surface and the endoscopy channel after the two cleaning procedures were cleaned (P>0.05). There was no significant difference in the ATP values between the gastroscope surface and the endoscopy channel after the two cleaning procedures were cleaned (P>0.05). However, the cleaning time of the test group was significantly less than that of the control group (P<0.01). Conclusion Both cleaning procedures can effectively and thoroughly clean the gastroscope. However, while ensuring the quality of the gastroscope cleaning, the test group can significantly reduce the gastroscopy cleaning time, improve work efficiency, and save labor costs, which is worth promoting.