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Abstract Introduction Surgery requires a high degree of precision, speed, and concentration. Owing to the complexity of the modern world, traditional methods cannot meet these requirements. Therefore, in this study, we investigated students’ diagnostic skills in the Operating Room in the context of surgical instruments by using gamification of surgical instruments and a crossover design. Method The study design was a multi-institutional quasi-experimental crossover and involved a three-arm intervention (with gender-specific block randomisation: Group A, B, and C) with a pre-test and three post-tests. A total of 90 students fell into three groups of 30 participants each. The surgical sets were learned for one semester through game-based instruction and traditional teaching, and then three OSCE tests were administered with time and location differences. Using one-way ANOVA, OSCE results were compared in the game, traditional, and control groups. The effectiveness of the intervention was tested in each group by repeated measures. Result The pretest scores of all three groups did not differ significantly. In the OSCE tests, both groups, A and B, performed similarly. However, these tests showed a significant difference in grouping between training through games and training in the traditional way. There was no significant difference between OSCE tests 2 and 3 in the game-based training group, indicating that what was learned was retained, while in the traditional method training group, OSCE 3 test scores declined significantly. Furthermore, repeated measures showed the effectiveness of game-based training. Conclusion In this study, gamification has turned out to be very effective in helping learners learn practical skills and leading to more sustainable learning.