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Healthcare professionals who are confronted with breaches in infection control practices often do not speak up.Likelihood to speak up is affected by profession and by hierarchy within profession.Risk to patients and past voicing behaviors are key drivers for speaking up.Resignation to speaking up was high and contributes to withholding voice.Hospital climate explains only a minor fraction of variance in speaking-up intentions.Speaking up by healthcare professionals (HCPs) is an important resource to reduce risks to patient safety. Due to complex tradeoffs, HCPs are often reluctant to voice their concerns. A survey investigated HCPs' likelihood to speak up.A cross-sectional survey study among HCPs in 5 Swiss hospitals addressed speaking-up behaviors, safety climate, and likelihood to speak up about poor hand hygiene practice described in a vignette. Likelihood to speak up was analyzed using a multilevel regression model.Of surveyed HCPs (n = 1217), 56% reported that they would speak up to a colleague with poor hand hygiene practice. Nurses as compared to doctors rated the situation as more realistic (5.25 vs 4.32, P < .001), felt more discomfort with speaking up (4.00 vs 3.34, P < .001), and reported a slightly lower likelihood of speaking up (4.41 vs 4.77, P < .001). Clinical function (hierarchy) was strongly associated with speaking-up behavior (P < .001). Higher risk of harm to the patient (P < .001) and higher frequencies of past speaking-up behaviors (P = .006) were positively associated with the likelihood to speak up. Higher frequencies of past withholding voice (P = .013) and higher levels of resignation (P = .008) were both associated with a lower likelihood to speak up.Infection control interventions should empower HCPs to speak up about non-adherence with prevention practices by addressing authority gradients and risk perceptions and by focusing on resignation.