A multicenter cross-sectional study was conducted, between May and July 2021 before any anti-Covid-19 vaccination program implementation, among 720 staff working in six hospitals in the province of North Kivu in the eastern Democratic Republic of Congo.The aim of the study was to determine the seroprevalence of anti-SARS-CoV-2 antibodies.Individual data on socio-demographic and professional parameters and wearing mask were collected on the basis of a standard form. A blood sample was taken for qualitative determination of anti-SARS-CoV-2 antibodies using immuno-chromatographic “Panbio COVID-19 IgG/IgM Device (25T)” kits.The overall IgG/IgM sero-prevalence was 32.9% (n = 720). This seroprevalence among hospital staff was not significantly associated with their age, gender, professional category, department to which they were assigned in the hospital, or location in a rural or urban setting of their hospitals, nor to the systematic wearing of masks.Among hospital staff who reported contact with a Covid-19 patient, seroprevalence was twice as high at service on the workplace 32.6% (n = 282) [(PR (95% CI)], [2.30 (1.46 – 2.95)] (p = 0.001).In conclusion, the anti-SARS-CoV-2 antibodies seropositivity level among service providers in hospitals in North Kivu province in Eastern DRC is high and that contamination seems more than half as frequent in a professional hospital environment as at the level of the family unit.