Group B Streptococcus (GBS) is the leading infectious cause of neonatal morbidity and mortality. Although intrapartum antibiotic prophylaxis (IAP) strategies are effective in preventing GBS transmission from mothers to newborns, there are growing concerns about adverse effects, and the development of antibiotic resistance. GBS vaccines targeting the most virulent neonatal disease serotypes are currently under development and may be used during pregnancy. The objective of this study was to explore the key issues and concerns that would be associated with GBS vaccination during pregnancy from the perspectives of pregnant women and health care providers. Twenty-two women and 25 health care professionals in Alberta, Canada participated in 10 focus groups, each group ranging from 2 to 20 participants. Valuable information emerged from the focus groups about the factors that would affect acceptance of a maternal GBS vaccine. This information will be essential for health systems to consider in the introduction, promotion and delivery of such a vaccine. The data may help optimize education about GBS and a putative vaccine to pregnant women.