ABSTRACT Objective: Human Cytomegalovirus (CMV) is a member of the Herpesviridae family, with the ability to establish a long-lived latent infection. CMV infection causes problems in immunocompromised hosts undergoing organ and stem cell transplantation. The prevalence of CMV in adults varies in different geographic regions. The purpose of this study was to assess the prevalence of CMV in the adult population of Afghanistan, which did not have epidemiologic information for CMV infection. Methods: A total of 500 adults residing in main regional provinces of Afghanistan including Nangarhar, Herat, Mazar-i Sharif, Kandahar and Kabul in the age range of 25-70 years old were randomly selected to include in the study. Among the participants, 263 (52.6%) were female and 237 (47.4%) were male. The samples were tested for the presence of CMV IgM and IgG antibodies using chemiluminescence immunoassay on the Abbott Architect automated platform. Results: The seropositivity of CMV was found 99.79 % in Afghanistan. There were no significant differences in the prevalence CMV infection among the five regions. The seropositivity anti-CMV IgG positive rate in Kandahar, Kabul, Nangarhar and, Herat was determined as 100%.  The CMV IgG prevalence was 98.99% in Mazar-i Sharif. Anti CMV IgM was found 1.24% in Afghanistan. Conclusions: Our study shown that the seroprevalence is high in Afghanistan. Because of the high frequency of seropositivity in general population in Afghanistan, the approaches for preventing CMV reactivation need to be developed.  J Microbiol Infect Dis 2019; 9(2): 78-82.