Background: Monkeypox disease (MPOX) recently re-emerged in May 2022, causing international outbreaks in multiple non-endemic countries. This study demonstrates a novel comparison between the knowledge and perceptions of Saudi healthcare workers (HCWs) and the general public regarding MPOX. Methods: An online survey, conducted from 27 May to 5 June 2022, assessing participants’ MPOX and monkeypox virus (MPV) knowledge in terms of transmission, vaccination, isolation precautions, and their attitudes toward seeking more information. Results: A total of 1546 members of the public and 1130 HCWs completed the survey. Briefly, 61.3% of the public and 74.2% of HCWs showed interest in seeking more information about MPOX. Both groups had average overall mean MPOX knowledge scores. Members of the public holding university degrees and those showing high levels of worry regarding MPOX had significantly higher knowledge scores. However, HCWs showed a poor vaccination knowledge score, while only 57% recognized that MPOX can present similarly to COVID-19 in the early stages. Female HCWs and those with high self-rated MPOX awareness had significantly high knowledge scores. HCWs in secondary and tertiary centers had significantly higher knowledge scores. Conclusion: Both groups showed a decent attitude in terms of seeking more MPOX knowledge, which correlated positively with their worry about and awareness of the disease. These observations are mostly as a consequence of the ongoing COVID-19 pandemic, which encouraged the public and HCW to acquire more information about any novel emerging disease. Policymakers should make the most of this attitude in their awareness campaigns to prevent the spread of the disease and encourage vaccination in cases where it is needed. The knowledge gaps among HCWs were most evident in terms of clinical presentation and vaccinations; this problem needs addressing if we are to avoid further emerging MPOX cases.