Background: Saudi Arabia is considered the epicentre of MERS-CoV. Since 2012, a total of 1844 cases of MERS-CoV have been reported. A recent cluster, with 8 cases, has been reported in Najran region in the south-western region of the country. This analysis of data from that region on health care workers (HCWs) awareness and attitudes towards the prevention and control of MERS-CoV may be useful when planning health education programmes about this emerging infectious disease. Aims: We aimed to investigate the knowledge and attitude of HCWs toward MERS-CoV in south-western Saudi Arabia. Methods: This cross sectional study was conducted on HCWs in primary health care centres and hospitals at Najran. A questionnaire containing 14 knowledge and 8 attitude items was completed by all 870 participants. Results: Overall, > 80% of HCWs were aware about MERS-CoV etiology, mode of transmission, risk factors, and signs and symptoms. Knowledge scores revealed 51% of participants had sufficient knowledge. Physicians and nurses had significantly better knowledge compared with other HCWs (P = 0.001). Participants who worked at institutions with established infection control programme scored significantly better on knowledge questions (P = 0.001). Concerning attitude, > 70% of HCWs exhibited a positive attitude toward MERS-CoV. Conclusion: the HCWs in Najran region showed a high level of knowledge and positive attitude toward MERS-CoV. There was a noticeable difference in knowledge level between different professions. Periodic educational interventions and professional campaigns are still needed. Any interventions should be directed towards the non-physician and non-nursing professions. Keywords: MERS-CoV; health care workers; knowledge; attitudes, infection control Citation: Asaad A; El-Sokkary R; Alzamanan M; El-Shafei M. Knowledge and attitudes towards Middle East respiratory syndrome-coronavirus (MERSCoV) among health care workers in south-western Saudi Arabia. East Mediterr Health J. 2020;26(4):435–442. https://doi.org/10.26719/emhj.19.079 Received: 19/05/18; accepted: 20/09/18 Copyright © World Health Organization (WHO) 2020. Open Access. Some rights reserved. This work is available under the CC BY-NC-SA 3.0 IGO license (https://creativecommons.org/licenses/by-nc-sa/3.0/igo).