Hepatitis B virus (HBV) is one of the viral diseases that is threatening the global health. This study was carried out to analyze knowledge, perception of hepatitis B virus and its vaccine uptake among residents of ondo state, south west of Nigeria. The study was a descriptive survey that involved the use of questionnaire to facilitate the sampling and interview of respondents. This included recruiting residents who gave their consent in three communities (Ile-Oluji, Ondo and Owo) at three different senatorial districts (south, central and North) of the state (one community per senatorial district). Four hundred and fifty consented to participate in the study and were selected. A validated self-administered questionnaire was used for data collection. Descriptive statistics and Chi-square tests were used to analyze the data. It was observed that the Knowledge of the subject matter needed much improvement in all the towns, especially the undeveloped ones in the state on awareness, perception and knowledge of the virus and the availability of its vaccine uptake.