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Objectives: The aim of this study was to investigate the prevalence of Group B Streptococcus (GBS) colonization in pregnancies between 35 and 37 weeks of gestation and to compare the effectiveness of polymerase chain reaction (PCR) method with gold standard technique of culture in antenatal GBS screening., Material and Methods: Vaginal and rectal swabs of a total of 106 pregnant women between 35th and 37th weeks of gestation, who were admitted to our clinic between January 2022 and August 2022, were evaluated using culture and PCR method. The prevalence of GBS was estimated. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of the PCR method were analyzed., Results: The prevalence of GBS was 10.4% and 21.69% using the culture and PCR method, respectively. Compared to the culture, the sensitivity, specificity, PPV, NPV and accuracy of PCR were found to be 100%, 87%, 47%, 100%, and 88%, respectively., Conclusions: This study results suggest that the PCR method is a simple, effective and fast method with high sensitivity, specificity, PPV, and NPV in antenatal GBS screening.