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Background: Training on endoscope reprocessing has been annually implemented for health practitioners working in endoscopy units of hospitals performing national gastric or colorectal cancer screening across the country, as part of the endoscopy quality improvement project. This study aimed to identify the effectiveness of endoscope reprocessing training. Methods: In this cross-sectional study, education on endoscope reprocessing was implemented 18 times across the country, from June 2019 to November 2019. A total of 1,186 participants were included and 1,132 of them answered the survey questionnaire (response rate, 95.4%). Results: Of the study participants, 45.8% had previous experience in training on endoscope reprocessing, and 87.6% of them have adhered to the endoscope reprocessing guidelines. Experience of participation in endoscope reprocessing training was significantly associated with practical adherence to endoscope reprocessing guidelines (aOR, 6.55; 95% CI, 3.93 to 10.91). The satisfaction with and need for endoscope reprocessing training were 92.7% and 95.7%, respectively. The level of knowledge and intention to adhere to endoscope reprocessing guidelines were both high (4.65 [1-5]). Conclusions: Training on endoscope reprocessing currently provided at the national level could be effective for obtaining quality control for endoscopy. However, completion of the training by actual practitioners in charge of endoscope reprocessing within the national cancer screening system remains low; thus, strategies for more aggressive training participation for them should be developed at a national level.