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Purpose: The aim of this study was to examine the relationship between handwashing knowledge and handwashing compliance in nursing students and to identify the moderation and mediation effects of self-efficacy and perceived behavioral control on the relationship. Methods: The participants were 172 nursing college students who completed the self-report measures. Data were analyzed using t-test, ANOVA, and Pearson correlation coefficient, Multiple regression analysis was used to test moderation and mediation effects. Results: The score for knowledge of handwashing was higher in participants who had completed fundamentals of nursing practicum (t=-6.54, p<.001) and the score for compliance with handwashing was higher in participants who had completed clinical practicum (t=-3.81, p<.001) or fundamentals of nursing practicum (t=-5.97, p<.001). Self-efficacy showed a significant moderation effect (z=2.21, p=.027) but not a mediation effect while perceived behavioral control had a significant mediation effect but not a moderation effect on the relationship between knowledge of handwashing and compliance. Conclusion: Findings indicate that clinical practicum and fundamentals of nursing practicum were helpful to nursing students in acquiring knowledge and in practicing proper handwashing technique. These findings suggest that continuing and repetitive education programs on handwashing need to be enhanced and strategies to strengthen self-efficacy and perceived behavioral control need to be included in education programs. [ABSTRACT FROM AUTHOR]