Background: Cytomegalovirus (CMV) is a common virus that can infect individuals of all ages. In pregnant women, CMV infection can lead to complications such as congenital CMV infection, which can cause miscarriage, stillbirth, or significant fetal abnormalities. Objective: To evaluate the prevalence of CMV infection in women with and without a history of miscarriage in Diyala Province, Iraq. Methods: A total of 120 women were recruited and divided into three groups: 40 pregnant women with a history of miscarriage, 40 non-pregnant women with a history of miscarriage, and 40 healthy women without a history of miscarriage (control group). Blood samples were collected from each participant and analyzed for the presence of CMV-specific IgG and IgM antibodies using enzyme-linked immunosorbent assay (ELISA). Some positive results were retested to detect the MIE gene region IE-2 (UL122) by real-time PCR and the IE1 (UL123) gene by conventional PCR. Demographic data of study groups were recorded, and statistical analysis was performed using SPSS Version 29. Results: The serological analysis revealed the following prevalence of CMV IgG antibodies: 100% in both pregnant and non-pregnant women with a history of miscarriage, and 95% in the control group of healthy women without a history of miscarriage. For CMV IgM antibodies, the prevalence was 47.5% in pregnant women with a history of miscarriage, 40% in non-pregnant women with a history of miscarriage, and 7.5% in the control group of healthy women without a history of miscarriage. The PCR results showed that 9 of the samples were positive for both the UL122 and UL123 genes. Conclusion: The study indicates a high prevalence of CMV infection among women with a history of miscarriage in Diyala Province, Iraq. Routine screening for CMV in pregnant women, especially those with a history of miscarriage, is recommended to reduce potential risks and improve pregnancy outcomes.