OBJECTIVE: To assess outcomes and costs associated with around-the-clock point-of-care intrapartum group B streptococcus (GBS) polymerase chain reaction (PCR) screening. METHODS: Intrapartum PCR screening was implemented in 2010. Intrapartum PCR was compared with antenatal culture screening in an uncontrolled, single institution, preintervention and postintervention study. The study periods included 4 years before and 6 years after the intervention, commencing in 2006 and concluding in 2015. The primary outcome measure was rate of early-onset neonatal GBS disease. Secondary outcomes included length of stay, days of antibiotics, and costs. RESULTS: During the 4 years of antenatal culture screening, 11,226 deliveries were recorded compared with 18,835 in the 6 years of intrapartum GBS PCR screening, corresponding to 11,818 and 18,980 live births, respectively. During the antenatal culture period, 3.8% of term deliveries did not undergo GBS testing compared with 0.1% during the intrapartum PCR period ( P <.001). Between the two periods, the rate of proven early-onset GBS disease cases decreased from 1.01/1,000 to 0.21/1,000 ( P =.026) and probable early-onset GBS disease cases from 2.8/1,000 to 0.73/1,000 ( P <.001); the risk ratio for both was 0.25, 95% CI (0.14–0.43). Total days of hospital and antibiotic therapy for early-onset GBS disease declined by 64% and 60%, respectively, with no significant difference for average length of stay or antibiotic duration preintervention and postintervention. The yearly cost of delivery and treatment of newborns with GBS infection was reduced from $41,875±6,823 to $11,945±10,303 ( P <.001). The estimated extra cost to avoid one early-onset GBS disease was $5,819. CONCLUSION: Point-of-care intrapartum GBS PCR screening was associated with a significant decrease in the rate of early-onset GBS disease and antibiotic use in newborns. The additional PCR costs were offset in part by the reduction in early-onset GBS disease treatment costs.