Background: Although, GBS is a natural flora of the ano-rectal region, it may colonize vagina and many infants can be infected during the passage through the birth canal. It has emerged as a leading cause of neonatal infections and deaths. Objectives: To estimate the rate of recto-vaginal carriage of GBS among pregnant females, describe its antimicrobial susceptibility profile and to detect some of its virulence genes by multiplex PCR. Methodology: Vaginal, anorectal and neonatal throat swabs which were collected from two-hundred and fifty pregnant women were inoculated in Todd-Hewitt broth for 24 hours then inoculated on blood agar plates. Antimicrobial susceptibility testing for GBS isolates was done and its virulence genes (scpB, bca, rib and HvgA) were identified by PCR. Also, the relation between these virulence genes and antimicrobial susceptibility was studied. Results: Among 250 pregnant females, 36(14.4%) were identified as GBS carriers with exclusive vaginal and anorectal colonization rates of 4% and 10.4% respectively. All isolates were susceptible to penicillin, ampicillin, cefepime, cefotaxime, ceftriaxone, vancomycin and linezolid. On the other hand, 19.4%, 80.6%, 44.4% and 13.9% of GBS isolates were resistant to each of erythromycin and azithromycin, tetracycline, levofloxacin and clindamycin respectively. ScpB, rib, and Hvg-A genes were identified in 100%, 69.4% and 33.3% of GBS isolates respectively. None of them had the bca gene. Conclusion: Screening for GBS colonization of pregnant females is recommended and determination of virulence and different surface proteins would be relevant for better diagnosis and further possible formulation of a vaccine.