Herpes simplex virus (HSV) infections are among the most widespread sexually-transmitted diseases (STDs) all over the world. This study aimed to determine the prevalence and factors associated with HSV-2 infection among women and their newborns. A two-month cross sectional and prospective study took place at the District hospital of Deido in the town of Douala, Cameroon. A total of 90 mothers aged 17-42 years old attending this health facility because of pediatric consultation and vaccinations of their newborns were included in the study. Blood samples were obtained from each mother and newborn, centrifuged and obtained sera were used to look for Immunoglobulin against HSV using ELISA-based method. In addition, complementary data of mothers and their newborns were documented. Mean age of mother and newborns was 28.9 ± 3.5 years old and 5.2 ± 4.1 months respectively. The prevalence of Herpes simplex virus type 2 was 90% (95% CI:  83.8 – 96.2) and 74.7% (95% CI: 65.7 – 83.7) in mothers and their babies respectively. The rate of postnatal HSV-2 transmission was of 10.3% (95%CI = 82.4% – 97.0%). Neonates aged 1-3 months and 4-6 months was 31 (OR = 30.7; 95%CI = 5.7 – 163.7; P-value < 0.0001) and 7 (OR= 6.9; 95%CI =1.2 – 37.3; P-value < 0.0001) times more at risk for HSV-2 infections than older ones. This study pointed out a high prevalence of Herpes simplex Virus type-2 in mothers and their babies. Clinicians should be aware of including HSV infections in the differential, how recognize and properly treat neonatal HSV infections with the ultimate aim to efficiently mitigate mother-to-child transmission.