Herpes simplex virus (HSV) infection is one of the worldwide infections. HSV-1 causes herpes labial and HSV-2 is the main cause of genital ulcer. Host's immune response to HSV defines by the presence of neutralizing antibodies to HSV in serum of newly and previously infected persons. A cross-sectional study was designed to determine the prevalence of HSV antibodies and potential risk factors of HSV infections among women attending some hospitals and health centers in Sana'a city based on type-specific serological assay. HSV-1 differs from HSV-2 by the type-specific enveloped glycoprotein G (gG). This is the base of typespecific ELISA, which is more accurate than ELISA using common antigens. In our knowledge, there are no previous published studies or recorded data regarding the seroprevalence of HSV in Yemen. Moreover, there is no guideline of diagnosis or treatment of HSV infections in women; especially pregnant women. A total of 315 women, including 235 sexually transmitted infections (STIs) patients, 48 herpes labial patients (HLP) and 32 apparently healthy women were enrolled in this study. Of the studied women, 307 were married and 8 were single, their ages range from14-49 years. Among married women; 53 were pregnant. The total seroprevalence was 2.5% for HSV IgM, 99.4% for HSV-1 IgG and 5.1% for HSV-2 IgG. There were statistically non-significant association between herpes infections and age, size of family, educational level, residence, marital status, pregnancy, abortion, blood transfusion and contraceptive use. In conclusion, the prevalence of HSV-1 antibodies was much higher than that of HSV-2. The prevalence of HSV-1 antibodies was higher than those reported in other countries which could be due to poor hygienic conditions in Yemen. The prevalence of HSV-2 antibodies was lower than those reported in other studies; which is probably due to the differences in sexual practices among these cultures.