We assessed the quality of hand hygiene among healthcare workers at a pediatrics hospital in Rio de Janeiro, Brazil. Hand hygiene was performed in 491 (34%) of 1,455 opportunities. Of these hand hygiene events, correct performance was observed in only 173 (35%). Multivariate analysis revealed that correct performance of hand hygiene was associated with the use of an alcohol-based product and a lack of jewelry (for all events) and employment in an infirmary with a comparatively higher ratio of nurses to patients (for events involving nurses).