This study aimed to determine the seroprevalence of Cytomegalovirus virus (CMV) infection among pregnant women in Ad Dhale'e city, Yemen. A cross-sectional study was conducted among 130 pregnant women attended for routine antenatal care in Al-Nasser hospital in Ad-Dhale'e city. Serum was obtained and tested for seropositivity of CMV IgG and IgM using electro-chemiluminescence immunoassays  (ECLIA) test. Of 130 blood sample, 122 (93.8 %) and 5 (3.8%) were identified with CMV IgG and CMV IgM seropositive respectively. While, 8 (6.2%) were seronegative (susceptible) to CMV infection. Statistically significant association (P< 0.05) was found between CMV IgG seropositivity and age. This study concluded that there was high seroprevalence of cytomegalovirus infections among pregnant women in Ad-Dhale'e city. Routine antenatal screening of pregnant women for CMV infection should be considered. Further studies with more sample size using advanced methods is recommended.