The purpose was to compare the prevalence of healthcare-associated infections (HAI) in the Swedish point prevalence survey with an assessment using the European Centre of Disease Prevention and Control (ECDC) definitions of HAI. A total of 1247 patients were included from three Swedish hospitals. The prevalence of HAI was higher in the Swedish survey as compared to when using the ECDC definitions. The correspondence between results according to Swedish and ECDC protocols was better in Region Västerbotten than at Karolinska University Hospital. In Västerbotten, but not at Karolinska University Hospital, quality control was performed on collected data in the Swedish point prevalence survey. The study highlights the importance of expert knowledge of HAI and quality control to obtain valid survey results.