ABSTRACT Objective: build and validate an educational video as a strategy to encourage adherence of nursing professionals to Standard Precautions. Methods: in the first phase, a descriptive study was conducted with 197 nursing professionals from two hospitals in Cuiabá, Mato Grosso, Brazil, to identify knowledge and factors affecting adherence to Standard Precautions; data were collected using the Questionnaire for Knowledge and Compliance with Standard Precaution (Portuguese version). In the second phase, a methodological study and an educational video were developed with the results obtained in the first phase, structured according to Nola Pender’s Revised Health Promotion Model. The Delphi technique was used in video validation, a process involving 13 specialists. Results: knowledge of nursing professionals about Standard Precautions ranged from 6.1% to 99%. The major deficits were related to the objectives and purposes of Standard Precautions. Adherence ranged from 36.0% to 95.4% and was low regarding the proper use of some personal protective equipment, recapping of used needles, hand washing after removing gloves, and procedures after exposure to biological materials. The script was validated in two rounds, reaching 82.3% and 92.8% of consensus, respectively, and the video was validated in the third round, with a 92.3% consensus. Conclusion: lack of knowledge about the objectives and purposes of Standard Precautions was observed among nursing professionals, and many of them do not fully adopt them. The video was considered valid by the specialists and can be used to expand knowledge and, consequently, encourage adherence of nursing professionals to Standard Precautions.