BACKGROUND: Vaccination is an important method to address the monkeypox epidemic. We aimed to analyze the knowledge of healthcare workers (HCWs) about human monkeypox and their attitudes toward vaccination.MethodsWe searched PubMed, Embase and Web of Science for articles and performed a meta-analysis using Stata 14.0 with a random-effects model. This study was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. RESULTS: A total of 34 studies with 43,226 HCWs were included in this meta-analysis. The results showed that 54 % (95 % CI: 0.39-0.69) of the HCWs were willing to be vaccinated against monkeypox, and only 40 % (95 % CI: 0.29-0.50) of the HCWs had good knowledge of monkeypox. By analyzing the vaccination history of HCWs, we found that history of smallpox vaccination did not significantly affect the willingness of HCWs to receive another vaccination (OR = 0.53, 95 % CI: 0.23-1.26), whereas HCWs who had been vaccinated with the influenza vaccine (OR = 2.80, 95 % CI: 1.29-6.11) or COVID-19 vaccine (OR = 3.10, 95 % CI: 2.00-4.81) showed greater willingness to receive the monkeypox vaccine. In terms of income, low-income HCWs were less willing to be vaccinated against monkeypox (OR = 0.69, 95 % CI: 0.54-0.89), whereas middle-income HCWs were more willing (OR = 1.45, 95 % CI: 1.04-2.02). Notably, although HCWs with education related to monkeypox had better knowledge of monkeypox than HCWs without education related to monkeypox, the difference was not statistically significant (OR = 1.83, 95 % CI: 0.80-4.18). CONCLUSIONS: Publicity and education on monkeypox should be strengthened so that more people, especially HCWs, can have a good understanding of monkeypox and be willing to be vaccinated.