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Immunization programmes have significantly contributed to public health prevention and control of many infectious diseases globally. Despite this, there is observed reluctance towards accepting vaccination by mothers of infants. This study determined levels of knowledge, perception, and attitudinal dispositions likely to be responsible for vaccination hesitancy observed among mothers in Ishara, Remo North Local Government Area Ogun State, Nigeria.This study was cross-sectional survey enrolling 395 consenting mothers from Ishara town by multistage sampling technique following ethical approval. Data collectionwas byvalidated paper-instrument with Cronbach’s alpha internal consistency between 0.81 and 0.79. Variables in the study were developed as weighted aggregate of responses for knowledge (14-point), attitudinal disposition (27-point), and perception (45-point) scales respectively, and analysed by the computer-assisted software SPSS version 21 assummaries of descriptive and inferential statistics at 5% level of significance.The results showed age of mothers was 25.90±6.70 years and 23.40% of mothers had no formal educational, while 61.7% had more than primary education. Majority (83.2%) of the mothers reported having a child under two years old at the time of the study. Their level of knowledge regarding immunization was average (8.07±2.192), so was perception score (30.78±7.56) and 41.91% of mothers expressed negative attitude towards vaccination and immunization. Majority (95%) of the mothers indicated having vaccinated their infants at birth.All variables correlated significantly with vaccination coverage (p<0.01). The study revealed links between knowledge, perception, and attitude with vaccination coverage. Better communication strategies is recommended to strengthen current immunization outcomesin rural areas.