Aim Global screening strategies for Group B Streptococcus (GBS) include risk- or culture-based methods to guide intrapartum prophylaxis. In Western Australia (WA), antenatal culture-based screening is routine; however, numerous culture methods exist, in addition to molecular methods. We aimed to assess the comparability of research and diagnostic screening approaches. Methods and Results Vaginal and rectal swabs were self-collected by pregnant women (n = 531) from King Edward Memorial Hospital, WA, in parallel to routine screening (35-37 weeks of gestation). Research methods involved culture (Strep B Carrot Broth (TM) and StrepB CHROMagar (TM)) and molecular methods (real-time PCR) and were compared to routine diagnostic screening (Lim Broth and Granada agar). Overall, GBS detection was comparable between research and diagnostic approaches (3-5% discrepancy, kappa = 0 center dot 76). Specificity/sensitivity of Carrot Broth((TM)) was 100%/89%, while that of CHROMagar((TM)) was 73%/100%, respectively. Direct PCR was unable to detect GBS in ~18% of specimens which were culture positive; however, it exhibited 100% specificity. Conclusions This clinical evaluation of GBS screening methods provides support for current practice. Significance and Impact of the Study Although CHROM was highly sensitive, further testing is recommended due to a high false-positive rate. Molecular assays are useful for rapid detection; however, low-titre samples may require additional enrichment prior to molecular analysis to improve sensitivity.