BACKGROUND: Healthcare-associated infections are a global concern and can be dependent on the infection prevention behaviours of nursing staff, which in turn can be influenced by working conditions. Qualitative studies are scarce, and a greater understanding of the relationship between working conditions and nursing staff behaviour is needed. AIM: The aim was to describe nursing staff's experiences and reflections on working conditions and infection prevention behaviours. METHODS: A qualitative study with semi-structured focus group interviews at four surgical units and two orthopaedic hospital units. Twenty-seven nursing staff (12 registered nurses and 15 assistant nurses) participated. Data was analysed using qualitative content analysis. RESULTS: We generated one theme: Navigating in a complex and shifting context. The result indicates that working conditions are sometimes inadequate, which can hinder the nursing staff's infection prevention behaviours. Even when working conditions seemed to be sufficient, hygiene routines could fail, since situations constantly arise in a hospital unit that are difficult to predict and regulate. DISCUSSION: This study highlights the complexities faced by nursing staff in maintaining infection prevention behaviours within the dynamic hospital work environment. While nursing staff are professionally obliged to comply with hygiene routines, organisational support is essential for fostering sustainable working conditions. A multi-tiered approach is needed, from first-line managers to decision-makers, to promote a supportive environment that sustains safe practices.