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Background & aim: The monkeypox virus (MPXV), an Orthopoxvirus family member, is the zoonotic agent that causes mpox (formerly known as monkeypox). The ongoing mpox pandemic has caused cases across continents involving 110 countries. This study aimed to assess mpox knowledge and its determinants among healthcare personnel. Methods: This cross-sectional study was conducted from June 6 to June 25, 2022, among 17 Arab countries. The self-administered questionnaire consists of 53 questions assessing the knowledge about the monkeypox virus. Results: In total, 5874 medical students and clinical doctors from 17 Arab countries participated in this study. Only 13.8% (n = 812) of respondents have ever received information about mpox during their studies in medicine. The mean knowledge score was 13.84, and the median score was 15 (range 1–34). More than half (51.3%, n = 3012) have heard about mpox before. A low proportion of the participants had a good level of knowledge on mpox. Only 11.7% of respondents had correctly identified the natural host and the incubation period of mpox. More than half (58.9%) were aware of the signs and symptoms of mpox. Few respondents (28%) believed that mpox and smallpox have similar signs and symptoms. Specialist doctors had higher knowledge of mpox (AOR = 2.96, 95% CI = 2.24–3.92, p