Rectovaginal colonization with Group B Streptococcus (GBS) of pregnant women can lead to serious infection of woman, fetus or newborn and end up fataly. In Czech Republic every pregnant woman is screened for GBS colonization. This article aims to determine prevalence and antibiotic resistance of this bacteria. Prevalence of colonization from 2013 to 2020 was studied. Since 2019 each GBS specimen was tested to antibiotic sensitivity. Mean prevalence turned out to be 19,77%. Resistence to penicillin (ampicilin) was 0,65%, clindamycin15,52%, erythromycin 20,45% and vancomycin 0%.