ObjectiveTo determine the acceptability of different methods of routine testing for group B Streptococcus (GBS) colonisation to pregnant women and health care professionals (HCPs), and to examine barriers and facilitators to their implementation. DesignQualitative study, embedded in a cluster randomised trial SettingFour NHS maternity units participating in the GBS3 Trial: two conducting routine antenatal enriched culture medium (ECM) testing; and two using routine rapid intrapartum testing.Sample39 women and 25 HCPs purposively sampled to ensure representation of women with various birthing experiences and different professions.MethodsWomen were interviewed approximately 12 weeks postpartum by telephone or online video call, using a semi-structured topic guide. HCPs were interviewed during the testing period of the trial. Interviews were transcribed for thematic analysis and summarised using the framework method. ResultsFour categories of interest emerged: (1) views of routine testing; (2) acceptability of the testing procedure; (3) preferences on the types of test; (4) improving the testing procedure. Routine GBS testing was well received by both women and HCPs. Most participants found the procedure acceptable and were willing to receive the offer of testing in the future. Preferences for different testing methods varied, with participants emphasising the importance of evidence and informed choice. ConclusionsRoutine GBS testing is acceptable to most women and HCPs. Areas for consideration and the practicalities of implementing testing in maternity services are highlighted.