Introduction: Maternal and neonatal tetanus remains a preventable public health concern despite the availability of effective immunization. Adequate knowledge of tetanus toxoid (TT) injection among antenatal mothers is essential to ensure complete immunization and prevent tetanus-related morbidity and mortality. Urban–rural disparities in maternal health awareness may influence immunization outcomes.

Methods: A comparative, non-experimental descriptive study was conducted among 100 antenatal mothers (50 urban and 50 rural) in selected areas of Bidar district, Karnataka. Participants were selected using non-probability convenient sampling. Data were collected using a structured knowledge questionnaire comprising demographic variables and 35 knowledge items related to tetanus toxoid immunization. Descriptive statistics were used to summarize data, while independent t-test and chi-square test were employed to compare knowledge levels and determine associations with selected demographic variables.

Results: Urban antenatal mothers demonstrated significantly higher knowledge levels than rural mothers across all domains (p < 0.05). Adequate knowledge (>75%) was observed in 84% of urban mothers and 56% of rural mothers. The mean overall knowledge score was higher among urban mothers (30.87 ± 2.64) compared to rural mothers (27.83 ± 2.07). Educational status showed a significant association with knowledge levels in both groups, while occupation was additionally significant among rural mothers.

Discussion: The findings reveal a clear urban–rural disparity in knowledge regarding tetanus toxoid immunization. Lower knowledge levels among rural mothers highlight the need for strengthened antenatal education and nurse-led counseling interventions.