(1) Background: We aimed to explore the feasibility of collecting intrapartum maternal Group B Streptococcus (GBS) colonization and immediate post-birth neonatal GBS colonization cultures for use in a larger trial and to identify cases of residual GBS, which were hypothesized to be less common in the probiotics group. (2) Methods: This sub-study added additional outcome measures to the parent study to identify intrapartum and neonatal colonization and compare between probiotic and placebo groups and to identify cases of residual GBS. Intrapartum maternal vaginal and rectal GBS cultures were collected at the time of admission to a hospital for labor and to give birth. Neonatal oral and nasopharynx GBS cultures were collected within 1–2 h of giving birth. (3) Results: Thirty intrapartum samples were collected; twenty-eight had complete data. The antepartum GBS results significantly predicted the intrapartum results (p = 0.005), with 86.7% of cultures remaining the same at both time points. There were four cases where the intrapartum GBS results were different to the 36-week antepartum cultures results. A case of residual GBS was identified in one probiotic group participant. None of the neonatal swabs were positive for GBS. No cases of EOGBSD occurred in infants born to the study participants. (4) Conclusions: Although the 36–37 week GBS results significantly predicted the intrapartum results, the utility for a larger research trial on probiotics to reduce antenatal GBS is unclear. Intrapartum GBS swab collection was feasible in a busy nurse, midwife, and physician practice. GBS was not recovered from neonatal oral and nasopharyngeal swabs. The pathways of neonatal GBS colonization require further study.