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• Among blood donors in May 2020, the overall seroprevalence of anti-SARS-CoV-2 antibodies was 1.4%. • Non-citizens had higher prevalence compared to citizens (OR 13.6, p = 0.001). • There was geographic variation in seroprevalence of anti-SARS-CoV-2 antibodies within Saudi Arabia. Serologic testing provides better understanding of SARS-CoV-2 prevalence and its transmission. This study was an investigation of the prevalence of antibodies to SARS-CoV-2 among blood donors in Saudi Arabia. To estimate the seroprevalence of anti-SARS-CoV-2 antibodies among blood donors in Saudi Arabia during the early phase of the COVID-19 pandemic. Serology results and epidemiological data were analyzed for 837 adult blood donors, with no confirmed SARS-CoV-2 infection, in Saudi Arabia from 20th to 25th May 2020. Seroprevalence was determined using electrochemical immunoassay to detect anti-SARS-CoV-2 antibodies. The overall seroprevalence of anti-SARS-CoV-2 antibodies was 1.4% (12/837). Non-citizens had higher seroprevalence compared with citizens (OR 13.6, p = 0.001). Secondary education was significantly associated with higher seroprevalence compared with higher education (OR 6.8, p = 0.005). The data showed that the highest seroprevalence was in Makkah (8.1%). Uisng Makkah seroprevalence as the reference, the seroprevalence in other areas was: Madinah 4.1% (OR 0.48, 95% CI 0.12−1.94), Jeddah 2.3% (OR 0.27, 95% CI 0.31−2.25), and Qassim 2.9 % (OR 0.34, 95% CI 0.04−2.89) and these were not statistically different from seroprevalence in the Makkah region. At the early months of the COVID-19 pandemic in Saudi Arabia, the seroprevalence of antibodies to SARS-CoV-2 among blood donors was low, but was higher among non-citizens. These findings may indicate that non-citizens and less educated individuals may be less attentive to preventive measures. Monitoring seroprevalence trends over time require repeated sampling. [ABSTRACT FROM AUTHOR]