The novel Coronavirus Disease 19 (COVID-19) caused devastating effects globally, and healthcare workers were among the most affected by the pandemic. To mitigate this impact, healthcare workers were prioritized in COVID-19 vaccination globally and in Ghana. However, hesitancy by healthcare workers to receive the vaccination resulted in delayed control of the pandemic. In Ghana, vaccine acceptance rate among healthcare workers was estimated to be 39.3% in the pre-vaccine rollout period. Consequently, this study assessed uptake of COVID-19 vaccination and associated factors among healthcare workers in Ghana in the post-vaccine roll-out period.This was an analytical cross-sectional study that used a semi-structured questionnaire to collect data on COVID-19 vaccination uptake and influencing factors from randomly selected 256 healthcare workers in Ayawaso West Municipality, Ghana. Bivariable and Multivariable logistic regression was performed using IBM SPSS version 22 to identify predictors of vaccine uptake and a statistical significance was declared at p<0.05.More than three-fourths of participants 220 (85.9%) had received at least one dose of the COVID-19 vaccination, while 36 (14.9%) were hesitant. More than half 139 (54.3%) had adequate knowledge about COVID-19 vaccination and the majority 188 (73.4%) had positive perceptions about its effectiveness. Moreover, 218 (85.2%) of HCWs had a positive attitude towards COVID-19 vaccination. Positive attitude towards COVID-19 vaccination (AOR = 4.3; 95% CI: 1.4, 13.0) and high cues to action (AOR = 5.7; 95% CI: 2.2, 14.8) were the factors that significantly predicted uptake of COVID-19 vaccination among healthcare workers.COVID-19 vaccination among HCWs in Ghana is promising. However, hesitancy to receive the vaccination among a significant proportion of HCWs raises concerns. To ensure vaccination of all healthcare workers, interventions to promote vaccination should target key determinants of vaccination uptake, such as attitude towards the vaccination and cues to action.