This study assessed the association among knowledge, attitudes and uptake of COVID-19 vaccine in Malawi, examining demographic factors influencing these variables. The study employed a quantitative research design. It thus sampled 394 participants from Malawi's three districts of Zomba, Dowa and Nkhatabay. Results showed that 163 (41.4%) participants had low levels of knowledge of COVID-19 vaccine, 231 (58.6%) had high levels of knowledge, 237 (60.2%) had a positive attitude, and 156 (39.8%) had a negative attitude towards the COVID-19 vaccine. On vaccine uptake, the study found that only 29 (17%) male and 52 (23%) female participants had received the vaccine. Further, participants with low levels of knowledge and a positive attitude towards the vaccine were 5.9 times more likely (p-value = 0.001) to be vaccinated than those with low levels of knowledge and negative attitudes towards the vaccine. On the other hand, those with high knowledge and a positive attitude towards the vaccine were 8.2 times more likely (p-value < 0.001) to be vaccinated compared to those with low knowledge and negative attitudes towards the vaccine. The findings highlight the importance of vaccine-related knowledge and attitudes in shaping uptake and reveal disparities across demographic groups. To improve vaccination coverage in Malawi, targeted interventions focusing on enhancing COVID-19 vaccine knowledge, addressing attitudinal barriers, and countering misinformation are needed. Strategies should be tailored to reach populations with lower uptake, such as males, younger adults, and those with lower education levels. Strengthening public health messaging, engaging community leaders, and building trust in healthcare systems are crucial for promoting widespread acceptance and uptake of COVID-19 vaccines in Malawi.