BACKGROUND: Infectious complications are frequent among critically ill neonates. Hand hygiene is the leading measure to prevent healthcare-associated infections, but poor compliance has been repeatedly documented, including in the neonatal setting. Hand hygiene promotion requires a complex approach that should consider personal factors affecting healthcare workers' attitudes. OBJECTIVE: To identify beliefs and perceptions associated with intention to comply with hand hygiene among neonatal healthcare workers. METHODS: An anonymous, self-administered questionnaire (74 items) based on the theory of planned behavior was distributed to 80 neonatal healthcare workers to assess intention to comply, attitude toward hand hygiene, behavioral and subjective norm perceptions, and perception of difficulty to comply. Variables were assessed using multi-item measures and answers to 7-point bipolar scales. All multi-item scales had satisfactory internal consistency (alpha > 0.7). Multivariate logistic regression identified independent perceptions or beliefs associated with a positive intention to comply. RESULTS: The response rate was 76% (61 of 80). Of the 49 nurses and 12 physicians responding, 75% believed that they could improve their compliance with hand hygiene. Intention to comply was associated with perceived control over the difficulty to perform hand hygiene (OR, 3.12; CI95, 1.12 to 8.70; P =.030) and a positive perception of how superiors valued hand hygiene (OR, 2.89; CI95, 1.08 to 7.77; P =.035). CONCLUSION: Our data highlight the importance of the opinions of superiors and a strong perceived controllability over the difficulty to perform hand hygiene as possible internal factors that may influence hand hygiene compliance.