Introduction: Hand hygiene plays a huge role in removing hospital infections. The aim of this study was to explore the nurses’ viewpoints about the factors affecting hand hygiene compliance. Methods: In this qualitative content analysis study, the data were collected through purposive sampling and semi-structured interviews with 15 nurses. Interviews were audio-recorded and transcribed verbatim. Thematic analyses were conducted using Lundman and Graneheim’s method. Results: Six themes were identified, including the facilitator and barriers to compliance with hand hygiene on personal, interpersonal, and organizational levels. One theme was personal facilitator, with categories of facilitating the cognition and adherence to values. Personal barriers included cognitive obstacles, attitudinal barriers, and physical barriers. The interpersonal facilitators included supportive social climate and appropriate culture building. The interpersonal barriers involved inappropriate culture building and being under pressure. The organizational facilitators were strong leadership style, good managerial support, and competent staff evaluation; the last theme was organizational barriers with categories of poor leadership style, ineffective staff development, inconsistency in organizational policy, and incompetent staff evaluation. Conclusion: This study adopted an integrated approach to examining the factors affecting the nurses’ hand hygiene compliance. It is recommended that future interventions should consider the differences at individual, interpersonal, and organizational levels and developed a tailoring approach.