ObjectiveTo evaluate obstetric healthcare provider knowledge regarding the prevention of group B streptococcal disease in South African infants.MethodsQuestionnaires exploring knowledge, attitudes and beliefs around group B streptococcal prevention were administered to consenting doctors and maternity nurses in a tertiary academic hospital. Qualitative assessments (focus groups) were undertaken with junior doctors and nurses.Results238 participants completed the questionnaire: 150 (63.0%) doctors and 88 (37.0%) nurses. Overall, 22.7% of participants correctly identified the risk-based prevention protocol recommended at this hospital. Most doctors (68.0%) and nurses (94.3%) could not correctly list a single risk factor. A third of doctors did not know the correct antibiotic protocols, and most (80.0%) did not know the recommended timing of antibiotics in relation to delivery. Focus group discussions highlighted the lack of knowledge, awareness and effective implementation of protocols regarding disease prevention.ConclusionsOur study highlighted knowledge gaps on the risk-based prevention strategy in a setting which has consistently reported among the highest incidence of invasive group B streptococcal disease globally. In these settings, education and prioritization of the risk-based intrapartum antibiotic strategy is warranted, but an alternative vaccine-based strategy may prove more effective in preventing invasive group B streptococcal disease in the long-term.