Streptococcus agalactiae or Group B streptococcus (GBS) is among the leading life-threating infectious agents in neonates. It is also known to cause disease in pregnant women and immunocompromised patients. The aim of the current study was to determine GBS prevalence among pregnant and non-pregnant women in Ismailia, Egypt. Four-hundred sixty-two vaginal swabs were collected from pregnant and non-pregnant women. A total of 27.9% women were found as positive GBS carriers. In respect to pregnant participants, 26% pregnant women were defined as asymptomatic GBS carriers versus 35.5% among non-pregnant participants. Age, parity, history of previous abortion or preterm labor were not significantly associated as potential risk factors for GBS carriage.