Background: Group B streptococcus is a leading infectious cause of morbidity in newborns and causes substantial disease in elderly individuals. Guidelines recommended antenatal culture-based screening as the optimal method for identifying chemoprophylaxis candidates. Objectives: To assess the primary care physicians' knowledge about group B streptococcus screening in pregnant women and to identify the barriers that prevent primary care physicians from screening as recommended. Methods: A cross-sectional study conducted at the Al-Wazarat healthcare center, Prince Sultan Military Medical City, Riyadh, Saudi Arabia. A self-administrated questionnaire was distributed to 89 physicians asking about their knowledge and barriers regarding group B streptococcal infection High Vaginal Swab (HVS) screening. Results: The GBS screening average knowledge is 74.0% distributed on 11.2% scored poor, 31.5% good, and 57.3% excellent level of knowledge. Eight physicians' attitude was positively agreeing with GBS screening statement with lowest agreement was in performing screening with primary care setting (76.4%). The barriers for GBS screening are system and protocol (52.8%), lack of training (46.1%), lack of tools (24.7%), fear of consequences (14.6%), and other barriers (16.9%). Regression analysis showed that in comparison to physicians younger than 30 years, physicians aged 30 -39 years has higher average by 9.4% (p-value = 0.039), 40 -49 years have higher knowledge with average 10.3% (p-value = 0.036), 50 years or older have higher knowledge on average 8.9% (p-value = 0.040) Conclusion: Education programs regarding GBS screening in pregnancy are needed to improve the primary care physicians' knowledge in order to prevent inevitable complications of GBS infection in pregnancy.

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