Abstract : Background and Aim: Hepatitis B virus (HBV) infection is a leading cause of morbidity and mortality worldwide. Healthcare workers are a high-risk group. Vaccination is the most effective defense against the virus. This study investigated the uptake of Hepatitis B vaccination among healthcare workers at the Federal Teaching Hospital, Ido-Ekiti, Nigeria. Methods: A structured questionnaire was used to collect the socio-demographic data and responses of 100 consenting clinical staff of the institution on the subject matter and the data was analyzed using the SPSS version 21.0 software package. Results: A total of 100 respondents participated in the study; 40(40.0%) males and 60(60.0%) females, with a male to female ratio of 1:1.5. The age range of the participants was 20 – 59years with a mean(±SD) of 38.3±11.0 years. Fifty-eight(58%) of the study participants had received at least a dose of the Hepatitis B vaccine while the remaining 42(42%) of the participants had not received any dose of the vaccine. The number of doctors who had received the Hepatitis B vaccine was significantly higher than that of non-doctors(p = 0.015). Age above 40 years, female gender, Good knowledge of Hepatitis B and being a doctor are positive predictors of vaccine uptake. Conclusion: Hepatitis B vaccine uptake rate in this study is low when compared to similar studies in Nigeria and the vaccination completion rate in this study is even much lower.