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Acceptance of coronavirus disease 2019 (COVID-19) vaccination appears as a decisive factor necessary to control the ongoing pandemic. Healthcare workers (HCWs) are among the highest risk groups for infection. The current study aimed to evaluate COVID-19 vaccine acceptance among HCWs in Kuwait, with identification of the psychological determinants of COVID-19 vaccine hesitancy. The study was conducted using an online anonymous survey distributed between 18 March 2021 and 29 March 2021. The sampling strategy was convenience-based depending on chain-referral sampling. Psychological determinants of COVID-19 vaccine acceptance were assessed using the 5C subscales and the Vaccine Conspiracy Beliefs Scale (VCBS). The total number of study participants was 1019, with the largest group being physicians (28.7%), pharmacists (20.2%), dentists (16.7%), and nurses (12.5%). The overall rate for COVID-19 vaccine acceptance was 83.3%, with 9.0% who were not willing to accept vaccination and 7.7% who were unsure. The highest rate for COVID-19 vaccine acceptance was seen among dentists (91.2%) and physicians (90.4%), while the lowest rate was seen among nurses (70.1%; <i>p</i> < 0.001). A higher level of COVID-19 vaccine hesitancy was found among females, participants with a lower educational level, and HCWs in the private sector. A preference for mRNA vaccine technology and Pfizer-BioNTech COVID-19 vaccine was found among the majority of participants (62.6% and 69.7%, respectively). COVID-19 vaccine hesitancy was significantly linked to the embrace of vaccine conspiracy beliefs. The highest 5C psychological predictors of COVID-19 vaccine acceptance were high levels of collective responsibility and confidence, and lower levels of constraints and calculation. The VCBS and 5C subscales (except the calculation subscale) showed acceptable levels of predicting COVID-19 vaccine acceptance based on receiver operating characteristic analyses. The participants who depended on social media platforms, TV programs, and news releases as their main sources of knowledge about COVID-19 vaccines showed higher rates of COVID-19 vaccine hesitancy. An overall satisfactory level of COVID-19 vaccine acceptance was seen among HCWs in Kuwait, which was among the highest rates reported globally. However; higher levels of vaccine hesitancy were observed among certain groups (females, nurses and laboratory workers, HCWs in the private sector), which should be targeted with more focused awareness programs. HCWs in Kuwait can play a central role in educating their patients and the general public about the benefits of COVID-19 vaccination to halt the spread of SARS-CoV-2, considering the high rates of vaccine hesitancy observed among the general public in Kuwait and the Middle East.