The implementation and sustainability of evidence-based practice (EBP) is a complex process. Best practice guidelines (BPGs) can facilitate the translation of knowledge from theory into practice, but they require working groups to adapt recommendations and encourage adherence to them. The aim of this study was to identify facilitators and barriers of BPGs in the Registered Nurses' Association of Ontario®, within the framework of Best Practice Spotlight Organisations® (BPSOs®), in a hospital setting. Methods: This study was conducted using the modified Delphi method (two rounds) with a cohort of BPSO® Best Practice Champions. The facilitators and barriers included in the first round were identified through a bibliographic search. The degree of concordance was determined in the second round using the highest mean resulting values. Results: A total of 44 nurses responded, with a mean age of 42.57 ± 8.563. The facilitators included the following: work environment; working in a hospital that encourages research; and dedicating time to EBP (p < 0.001). The barriers included the following: excessive workload; professional mobility (p < 0.001); and lack of time (p = 0.002). Implications for practice: As such, it is necessary to approach human resource management by implementing new policies that guarantee systemic sustainability. The BPSO® programme is an excellent framework for promoting the translation of theoretical knowledge into practice and effecting institutional change.