question: OBJECTIVES: The National Institute for Infectious Diseases ‘Prof. Dr. Matei Balș’ was the designated centre for managing Ebola alerts in Romania during the 2014 African outbreak. We surveyed Ebola knowledge, attitudes and perceptions (KAP) among the institute's healthcare workers. STUDY DESIGN: This was a cross-sectional survey. METHODS: The study consisted of a self-administered paper-based anonymous questionnaire that included 24 closed-item questions and two scales of personal concern. RESULTS: Respondents were generally well informed; compared to nurses, doctors recorded a 1.9-fold higher rate of correct responses regarding Ebola transmission (P < 0.001), but both nurses and doctors correctly identified Ebola's aetiological agent. Nurses perceived higher personal (P = 0.008) and family (P < 0.001) risk than doctors. Respondents reporting high perceived risks were more likely to be less informed about Ebola (P = 0.019) and its prevention options (P = 0.033). Males were 6.7-fold more likely to volunteer than females (P = 0.001) and so were graduates of higher rather than lower education (1.5-fold more likely, P = 0.017) and doctors than nurses (1.7-fold more likely, P = 0.018). The institute ranked first among sources of information on Ebola; respondents who had received Ebola training in the institute 2 years previously were 1.2–1.3 times more likely to correctly identify transmission routes. CONCLUSIONS: We have characterised KAP on Ebola disease among Romanian healthcare workers from a tertiary care hospital in Bucharest. Nurses, specialist physicians and laboratory personnel may need more frequent retraining than residents and senior physicians.