Background: Healthcare workers' (HCWs) preparedness to respond to pandemics is critical to containing disease spread. Low-and middle-income countries, however, experience barriers to preparedness due to limited resources. In Ghana, a country with a constrained healthcare system, we examined HCWs' perceived preparedness to respond to coronavirus disease 2019 (COVID-19) and associated factors. Methods: The 472 HCWs completed questions in a cross-sectional self-administered online survey. Perceived preparedness was assessed using a 15-question scale (Cronbach alpha = 0.91) and summative scores were created (range = 0-45). Higher scores meant greater perceived preparedness, with scores ≥ 30 considered prepared. We used linear regression with robust standard errors to examine associations between perceived preparedness and potential predictors. Results: About 27.8% of HCWs felt prepared to respond to COVID-19. The average perceived preparedness score was 24 (standard deviation = 8.9). In multivariate analysis, factors associated with higher perceived preparedness were: training (β = 3.35, 95% confidence interval [CI], 2.01-4.69); having adequate personal protective equipment (PPE; β = 2.27, 95% CI, 0.26, 4.29), an isolation ward (β = 2.74, 95% CI, 1.15, 4.33), and protocols for screening (β = 2.76, 95% CI, 0.95, 4.58); and good perceived communication from management (β = 5.37, 95% CI, 4.03, 7.90). When added to the model, perceived knowledge decreased the effect of training by 28.0%, although training remained significant, suggesting a partial mediating role. Perceived knowledge was associated with a 6-point increase in perceived preparedness score (β = 6.04, 95% CI, 4.19, 7.90). Conclusion: HCWs reported low perceived preparedness to respond to COVID-19. Training, clear protocols, PPE availability, isolation wards, and communication play an important role in increasing preparedness. Government and other stakeholders must institute interventions to increase HCWs' preparedness to respond to the ongoing pandemic and prepare for future pandemics.