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Background: Human herpesvirus 8 (HHV-8) is a critical causative agent behind Kaposi sarcoma (KS), an oncogenic disease with profound consequences in immunocompromised individuals. Studies suggested HHV-8 seroprevalence in healthy populations is uncommon, but comprehensive investigations within the Middle East region remain scarce. This study aimed to bridge this knowledge gap by meticulously assessing HHV-8 seroprevalence among healthy blood donors in Qatar, leveraging serological methodologies and PCR., Methods: We used sera samples collected from 621 healthy blood donors (median age = 36 years, IQR 30-43) from different nationalities residing in Qatar, mainly from the MENA region and Southeast Asia. All sera samples were tested for total anti-HHV-8 IgG antibodies using ELISA. The presence of lytic HHV-8 antibodies was confirmed by an immunofluorescence assay (IFA). Further, HHV-8 DNA was tested and quantitated by qRT-PCR., Results: ELISA detected anti-HHV-8 IgG total antibodies in 6.9 % [43/621, 95 %CI 5.2-9.2] of the tested samples. Subsequent testing by IFA revealed that 14 % [6/43, 95 %CI 3.6-24.3] of these anti-HHV-8 IgG were classified as HHV-8 lytic antibodies. This suggests that 0.97 % [6/621, 95 %CI 0.2-1.7] of these donors had a recent or ongoing active infection and viral replication. Only one seronegative Qatari blood donor had detectable HHV-8 DNA in his blood. No significant difference was observed between HHV-8 seropositivity and the demographic characteristics of the donors., Conclusion: Our study showed that HHV-8 prevalence in Qatar aligns closely with global reports. Moreover, our findings raise considerations regarding HHV-8's potential transmission via transfusion, which suggests the value of routine HHV-8 screening, particularly for immunocompromised patients vulnerable to KS., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)