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To identify and compare tools to assess Evidence-Based Practice (EBP) behaviour among healthcare professionals.Systematic review.MEDLINE, EMBASE, Cochrane Library, PsychInfo and CINAHL up to July 2011.Titles, abstracts and eligible full text articles were screened by two reviewers independently.Relevant data were extracted by one reviewer and checked by a second reviewer. Eligibility criteria for selecting studies: original studies among all healthcare professionals that described the development or use of EBP behaviour assessment tools.Of 19 310 identified articles, 172 studies were included. We identified 117 questionnaires, 10 interviews or focus groups, nine observational studies, 27 chart evaluations and nine studies used a combination of methods. Psychometric properties of the questionnaires used were reported in about half of the studies, in seven studies that assess a single EBM step and in six studies that assess a combination of EBM steps. One of these assessed all five steps of EBP.Valid and reliable EBP behaviour assessment tools are available. However, only one questionnaire validly assessed all five EBP steps, covering the entire concept of EBP.