Background and Aim: Group B Streptococcus (GBS) has been identified as a pathogen since 1970. This organism can cause poor outcome and death in neonatal period and also puerperal infection. In other countries, the prevalence of rectal and vaginal colonization of GBS is about 20-30%. In Iran there are different reports about the prevalence of GBS colonization. Diagnosis and treatment of GBS colonization can prevent mortality in neonatal period and morbidity during post partum period. The aim of this study was to determine the prevalence of GBS colonization and sensitivity to various antibiotics in women who were visited in prenatal clinic of Mahdieh Hospital.Materials and Methods: This descriptive study was carried out on 246 women at 35-37 weeks gestation in Mahdieh hospital in 2008. Vaginal & rectal samples were cultured in special medias. If there was positive gram, CAMP test was done to identify GBS, then their sensitivity to antibiotics were studied. Our findings were analyzed by SPSS software.Results: Prevalence of GBS in our study was 5.3% and there was no correlation with age, education, rupture of membrane, parity and nationality. But there was a poor relation between multiple gestation and positive culture. Antibiotic sensitivity was identified as a sequence of Erythromycin, Cephasoline, Clindamycin, Vancomycin, Ampicillin, and Penicillin.Conclusion: In our study, the prevalence of GBS was lower than other studies from Iran, and sensitivity of GBS to Penicillin was lower than other antibiotics.