Background: Maternal genital tract colonization with GBS during pregnancy is an important risk factor for neonatal GBS infections. Methodology: This prospective study was done to determine Group B streptococcus (GBS) colonization in pregnancy using selective and non-selective culture media. The total sample size was 300 pregnant women. In 200 women, vaginal and rectal swabs were collected and inoculated separately in non- selective blood agar media and in another 100 women, vaginal and rectal swabs were collected and inoculated separately in selective Todd-Hewitt broth media for GBS isolation. Results: The detection rate for GBS was more when Todd Hewitt Broth was used for initial inoculation than direct inoculation in blood agar. The overall GBS prevalence rate among pregnant women was 2.3% in our study. Conclusion: Our finding supports the current recommendations that inoculation in selective medium is fundamental to maximize GBS detection rates and should be employed by laboratories involved in screening of pregnantwomen.