Background:
The recent human monkeypox (MPOX) outbreak in non-endemic countries has raised concerns among public health authorities worldwide. Healthcare professionals (HCPs) play a decisive role during epidemics in transmitting accurate information to the public and motivating them to pursue protective behaviours, including immunisation.

Methods:
A cross-sectional survey-based study was conducted in the Czech Republic in September 2022 to evaluate MPOX-related knowledge and vaccination perceptions among HCPs. The study utilised a digital self-administered questionnaire (SAQ) which inquired about participants’ sociodemographic and anamnestic characteristics, perceived knowledge of MPOX, factual knowledge, and vaccination perceptions according to the health belief model (HBM).

Results:
A total of 341 participants were included in this study; most of them were females (88.9%), allied HCPs (89.4%), heterosexuals (87.1%), married (61.9%), and vaccinated against COVID-19 (91.2%). Only 8.8% of the participants agreed to receive vaccination against MPOX; 44.9% rejected it, while 46.3% were hesitant. While digital news portals (47.5%) and social media (25.8%) were among the most utilised sources of information about MPOX, the scientific journals (5.6%), ECDC (5%), and the U.S. CDC (1.5%) were the least common sources. The participants demonstrated suboptimal levels of factual knowledge, especially regarding MPOX vaccines (1.5 ± 1.2 (0–4)) and treatments (0.9 ± 0.9 (0–4)). Additionally, several misconceptions were detectable among the participants regarding topics such as the availability of effective vaccines and antivirals against MPOX, the risk of vertical transmission, and homosexual stigmatisation. The HBM indicated that the cues to action and perceived susceptibility were the most important constructs to predict MPOX vaccine acceptance.

Conclusions:
The findings of this study call upon public health practitioners and health policymakers in the Czech Republic to tailor dedicated educational campaigns should encounter the HCPs’ misconceptions about MPOX, and future studies should explore the prevalence and drivers of MPOX vaccine hesitancy among the general population.