Purpose: This study aimed to explore the knowledge and performance of sleep hygiene among nurses with shift work schedules and examine the influence on sleep disturbance. Methods: A total of 199 shift-work nurses from a tertiary hospital were included in the study. To examine the knowledge and performance of sleep hygiene, the participants were asked to respond to a self-reported survey. To assess sleep disturbance, the General Sleep Disturbance Scale was used. Descriptive statistics and multiple linear regression were applied using the SPSS/WIN 21.0 program to analyze the data. Results: Both the knowledge and performance of sleep hygiene among nurses with shift work schedules were measured at a moderate level. Nurses’ knowledge and the performance of sleep hygiene was not significantly associated with sleep disturbance, whereas age, experience with shift-working, and perceived health status were significantly associated. Conclusion: Strategies for providing more accurate information and motivating better sleep hygiene would help to enhance sleep hygiene in nurses with shift work schedules. Further studies examining the association of knowledge and the performance of sleep hygiene with sleep disturbances in nurses with shift work schedules are needed.