Objective To select a high-level disinfection method suitable for disposable portable bronchoscope, and to explore the application value of adenosine triphosphate (ATP) bioluminescence detection in the evaluation of disinfection effect. Methods Ten new disposable portable bronchoscopes were randomly divided into control group (n＝1) and 3 experimental groups (hypochlorous acid group ［n＝3］, peracetic acid group ［n＝3］, and low-temperature plasma group ［n＝3］). The control group did not receive any treatment. The hypochlorous acid group, peracetic acid group and low-temperature plasma group were disinfected respectively by hypochlorous acid immersion, peracetic acid immersion and low-temperature plasma sterilization after simulated clinical aspiration. The bronchoscopes in each experimental group were pretreated and decontaminated by incomplete immersion. The time of hypochlorous acid immersion, peracetic acid immersion, and low-temperature plasma sterilization was 3, 5, and 45 min, respectively. The disinfection effect was evaluated by ATP bioluminescence detection and bacterial culture. Results The results of ATP bioluminescence detection showed that the relative luminous unit (RLU) values of bronchoscopic lumen sampling solution in the hypochlorous acid group and peracetic acid group after 5 rounds of disinfection were between 16 and 179 (all values＜200), and the qualified rates of disinfection were 100% in both groups. After 2 rounds of disinfection in the low-temperature plasma group, the RLU values of bronchoscopic lumen sampling solution were between 675 and 4 532 (all values＞200), and the qualified rates of disinfection were 0. After 5 rounds of disinfection, the results of bacterial culture in the hypochlorous acid group and peracetic acid group showed that colony-forming units were between 0 and 6, and the qualified rates were 100%. After 2 rounds of disinfection, the results of bacterial culture in the low-temperature plasma group showed that colony-forming units were greater than 20, and the qualified rates were 0. There were no significant differences in the image quality, distal curvature or negative pressure attraction of the bronchoscopes between the experimental groups and the control group after each round of disinfection. Conclusion Both hypochlorous acid immersion disinfection and peracetic acid immersion disinfection can achieve a high level of disinfection without affecting the performance of bronchoscope after the pretreatment of disposable portable bronchoscope. The results of ATP bioluminescence detection are consistent with those of bacterial culture, and it can quickly evaluate the disinfection effect of disposable portable bronchoscope.