The epidemiology of herpes simplex virus (HSV) infections varies among populations depending on their demographic characteristics and exposure. This study describes the prevalence of HSV 1/2 IgG and IgM antibodies among individuals over a period of 5 years. A retrospective study was conducted to collect data on cases tested for HSV-1 and HSV-2 IgG and IgM antibodies for different medical conditions over five years between January 2014 and December 2018. 620 samples were tested for HSV 1/2 IgG and IgM during the study period. The total HSV seropositivity in the study population was 68% (422/620). The total seropositivity excluding children below 6 months of age was 65.3% (313/479). HSV-IgG seropositivity was significantly higher in married individuals (p<0.001, 95% CI 1.61–3.69). The HSV IgG seropositivity was significantly higher in children under the age of 6 months (N=109, 77.30%) than in children between 7 and 24 months old (27.6%) (Chi-square for linear trend, p<0.001), and it then tends to increase with age more than 24 months (Chi-square for linear trend, p=0.011). Eleven children showed laboratory evidence of recent HSV infection (6.2%) as indicated by HSV IgM antibodies and had diverse clinical conditions. HSV infection is highly prevalent in the Eastern Province of Saudi Arabia. Infection is most probably acquired during early childhood, and the tendency increases with age. However, a significant number of mothers are at risk of infection and transferring the infection to their fetuses.