We assessed the knowledge and attitude (K&A) toward Crimean-Congo haemorrhagic fever (CCHF) of occupationally at-risk healthcare workers (HCWs). A cross-sectional survey was performed in three referral hospitals in the Systan-Baluchestan and Isfahan provinces of Iran where CCHF is highly endemic. In all, 191/209 eligible HCWs were enrolled (response rate: 93%). All but 11 (5.8%) had heard of CCHF. The mean K&A scores of the respondents were 50.34% and 79.25%, respectively. The correlation between K&A was significant (correlation coefficient: 0.542; P<0.001). Being a physician, working in Isfahan (versus the relatively deprived Systan-Baluchestan) and relying on academic material rather than local media were independent factors significantly associated with more knowledge; higher education and laboratory staff with better attitude were also significant factors. Although HCWs showed the best K&A for preventive measures, only 44% wore gloves and masks for contact with CCHF patients and 22% failed to observe any safety measure. Those with a history of percutaneous contact (6.3%) had significantly lower knowledge scores (P=0.047). There is a need to establish professional education campaigns in highly endemic deprived areas in order to improve physicians' attitudes, encourage nurses' use of academic materials and increase the knowledge of less-educated HCWs.