Background: Sexually transmitted diseases are known to pose a significant public health threat. Herpes simplex virus type 2 (HSV-2) is a viral infection with high prevalence in Sudan, particularly among pregnant women. The aim of this study was to determine the seroprevalence of HSV-2 among pregnant women as well as their associated potential risk factors in Wad Madani-Sudan. Methods: In total, 92 pregnant women who attended the Maternity Hospital and Dr. Altigani Sedeeg Fertility Center at Wad Madani, AL-Gezira State, between April and August 2018 were included in this study. HSV-2 Immunoglobulins (IgG and IgM) were assayed by the enzyme-linked immunosorbent assay technique. Results: The seroprevalence of HSV-2 IgM and IgG were 35(38%) and 90(97.8%), respectively. A Higher prevalence of HSV-2 IgG 45(48.9%) and 13(14.1%) for IgM was detected in pregnant women aged 15-24 years old with a significant association between HSV-2 IgM and age distribution group (p-value=0.001).  HSV-2 IgM and IgG seropositivity were highest in the first trimester; 26(34.2%), and 75(81.5%), respectively. The highest rate of HSV-2 IgM was detected in 19 (20.7%) women with a history of abortion, while the highest rate of HSV-2 IgG was seen in 50 (54.3%) women without a history of abortion. The statistical analysis results did not reveal significant differences between gestational stages and history of abortion with the presence of HSV-2 antibodies. Conclusion: The prevalence of HSV-2 was significant among pregnant women in Wad Madani- Sudan. There was a strong association between HSV-2 IgM seropositivity and the age of pregnant women.