Background:This study aimed to determine factors affecting compliance with hand hygiene from the perspective of the health care team working in the PICU. Materials and Methods:We recruited 35 PICU health care workers, including nurses, attending physicians and medical students, selected by purposive sampling between February to November 2020. Data were collected using deep semi-structured interviews. Then analyzed through Graneheim and Lundman’s (2004) qualitative conventional content analysis method. All interviews were recorded and typed in the MAXQDA software and then analyzed by eight steps including writing interviews, determining the semantic units, text coding, matching codes with text, categorizing and developing categories, reviewing the categories, identifying the subthemes and reporting the findings. Results: 68.5% of the participants were female, with an average age of 32.7 years. Overall, 234 codes were extracted from the interviews and after the integration of similarities 4 main categories were finally extracted: (I) Meta-organizational factors including two subcategories of macro-health policy and sanctions and economic problems, (II) Organizational factors including three subcategories of incoherence in observation of rules, monitoring and evaluation, the prevailing attitude of the organization, (III) Individual factors including three subcategories of attitude of the health care team towards patient, skin reactions to hand sanitizers, concerns about their own health and health of their family, and (IV) Characteristics of the clinical environment including three subcategories of professional relationships, special conditions governing the pediatric ICU, and care facilities. Conclusion:Based on the results, four key themes including Meta-organizational factors, Organizational factors, Individual factors and characteristics of the clinical environment were obtained indicating factors affecting hand hygiene performance of the PICU health care team.