Background: Although numerous studies have stressed the importance of compliance with methicillin-resistant Staphylococcus aureus (MRSA) protocols with regard to cost reduction and a safer environment for health care workers and patients, an evaluation of the usability of the protocols themselves is lacking. In this study, we evaluated the usability and performance of those protocols. - Methods: The performance of MRSA protocols was examined in 5 Dutch hospitals by means of a questionnaire (n = 63), followed by a practical test (n = 50), in a stratified random sample of 3 types of health care workers (physicians, nursing staff, and cleaning personnel). The questionnaire consisted of constructs related to exposure to risk, risk perception, knowledge of and attitude toward the protocols and safety preventive measures, self-reported behavior, and social and organizational support. The practical test consisted of “what if” scenarios that simulate the actual use of the protocol as a guideline for solving infection problems. The health care workers were asked to verbalize their thoughts and actions while using the protocol. - Results: The questionnaire demonstrated adequate knowledge of and attitude toward the MRSA protocols. However, the practical test revealed that the majority of respondents had problems with the accessibility, comprehensibility, applicability, and acceptability of the protocols. Problems not only occurred because of unclear information about the preventive measures and a poor infrastructure but also because of preventive measures that conflict with principles in providing care. - Conclusion: The protocols do not reflect the practical needs of the health care workers. In view of the different decisions that health care workers have to take in various circumstances, it would be impracticable to use the same MRSA protocol for all hospital staff. Finally, the practical test provided more reliable results than the questionnaire.