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Abstract Introduction The advent of the effective COVID-19 vaccine was the most eagerly expected worldwide. However, this hope quickly became hesitation and denial in many countries, including Guinea. Understanding the reasons for low vaccine coverage is essential to achieving herd immunity leading to disease control. This study aimed to comprehend the facilitators and barriers to the acceptance COVID-19 vaccine in Guinea. Methods The survey focused on healthcare workers (HCWs) and the general population (GP) in 4 natural regions in Guinea from 23 March 2021 to 25 August 2021. We used the Fishbein integration model to study the behaviours of HWCs and GP regarding vaccination. A mixed cross-sectional study collected knowledge, attitudes, norms, and perceptions. Regression and thematic content analysis identified the main facilitators and barriers to vaccination. Results We surveyed 3547 HCWs and 3663 GP. The proportion of people vaccinated was 65% among HCWs and 31% among the GP. For HCWs: the main factors associated with vaccination against COVID-19 were as follows: absence of pregnancy AOR = 4.65 [3.23–6.78], being supportive of vaccination AOR = 1.94 [1.66–2.27] and being an adult AOR = 1.64 [1.26–2.16]. Regarding the GP, the following factors increased the odds of vaccination: absence of pregnancy AOR = 1.93 [CI 1.01–3.91], being favourable for vaccination AOR = 3.48 [CI 2.91–4.17], being an adult AOR = 1.72 [CI 1.38–2.14] and being able to get the vaccine AOR = 4.67 [CI 3.76–5.84]. Semi-interviews revealed fear, lack of trust, and hesitant perception of the government as potential barriers to vaccination. Conclusion This study suggests that beliefs and negative perceptions are potential barriers to vaccination against COVID-19 among HCWs and the GP. Policies should emphasise practical strategies to mitigate these barriers among young people and pregnant women. Lastly, there is a need to improve access to vaccines in the GP.