Background: Hand hygiene (HH) is the most effective means of preventing healthcare-associated infections (HAI). HH improvement strategies primarily focus on healthcare staff, often overlooking the significant contribution of caregivers to HAI risk. We sought to understand caregivers’ HH knowledge and practices to identify improvement opportunities. Methods: A self-administered survey was developed and distributed to families from June to August 2019; open-ended questions and Likert scales assessed caregivers’ perceptions and practices regarding HH at home and in hospital. HH compliance audits of caregivers entering and exiting inpatient rooms were performed in the same time period. Results: Among 81 caregivers surveyed, median patient age was 4.0 (IQR, 0.9–13.0) years. This was the first admission for 42 patients (53.8%). During this admission, 22 (27.2%) patients had been admitted for ≤1 day and 45 (55.6%) for >3 days. Caregivers reported good knowledge of HH practice, with strongly positive responses to knowledge of HH moments (94%) and proper technique (96%). Caregivers recognized that HH is required of hospital visitors (96%) to protect others (99%) and prevent illness in hospital (93%). Responses were less consistent for performing HH before entering a hospital room (83%), after exiting the room (70%), or after coughing or sneezing (65%). The attitudes of caregivers of children above 2 years were equivocal regarding expectations of their child to wash hands upon entering (40%), or exiting (41%) the hospital room. Multivariable modeling identified higher self-reported HH compliance in caregivers during first admission to hospital, compared to subsequent admissions (OR, 3.15; 95% CI, 1.11–9.65). Reported barriers to HH included hand irritation (27.2%) and perceived HH frequency (18.5%). At the time of survey completion, 62 caregivers (77%) reported not having received HH information during their child’s admission from a healthcare provider or volunteer. Information was most commonly gained from posters (75%) and information in the room (31%). Most caregivers (58.0%) reported that they would prefer to receive HH information in the first 24 hours of admission. Among 200 audits, overall caregiver compliance with HH was 9%; HH before entering the room was 7.2% compared to 11.2% after exiting (P = .33). Conclusions: Reported caregiver knowledge of HH was not reflected in audited practice. Fewer than 1 in 4 had received HH information from healthcare staff. HH education in the hospital environment within the first day of admission provides an opportunity for caregivers to improve compliance as partners in HAI prevention and safer pediatric care.