IntroductionCountries in sub-Saharan Africa with a high hepatitis B burden also have limited resources to identify underlying drivers of disease among key at-risk populations. To improve prioritization and strengthen prevention of mother to child transmission of HBV, it is imperative to understand disease awareness, knowledge and related factors among pregnant women.ObjectivesThis study assessed HBV disease awareness, knowledge and related factors among pregnant women in public health facilities in two regions with diverse HBV disease epidemiology.MethodsFrom October 2016 through December 2017, a random sample of 455 pregnant women attending antenatal clinics were surveyed to assess HBV awareness, knowledge and associated factors. Participants responded to an 18-item questionnaire with themes on HBV awareness, knowledge of disease signs and symptoms, transmission, prevention and misconceptions about the disease. Results were analysed in STATA (version 14.0).ResultsOf 455 participants enrolled, about two thirds reported having heard about HBV disease. By region, nearly half (47%) of participants from the central region, compared to only 16% from the north, reported that they had never heard of HBV. Region of residence had a moderating effect on the education- HBV awareness relationship. Only 162/455 (36%) of participants had adequate HBV knowledge. More than half 256/455 (56%) and 242/455 (53%) were not knowledgeable about horizontal and mother to child HBV transmission, respectively. About two thirds 298/455 (66%) and 281/455 (62%) believed HBV was spread via sharing of utensils and mosquito bites respectively. In multiple regression analysis, residing in the north, (PR=1.91(1.53 -2.38), p < 0.001) compared to central region and having a secondary education (PR=1.87(1.37 -2.55), p < 0.001) compared to primary were statistically significantly related to being knowledgeable about HBV.ConclusionWe demonstrated marked regional differences in HBV disease awareness and knowledge in this high HBV prevalence setting. However, most pregnant women displayed unacceptably low HBV knowledge and a significant proportion still hold misconceptions about HBV. Interventions to improve HBV prevention through antenatal education will need to be tailored to existing differences in comprehensive HBV knowledge.