Abstract Seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) IgG antibodies, using dried blood spots, was determined in October–November 2020, among residents and staff randomly selected from 20 nursing homes (NH) geographically distributed in Flanders, Belgium. Sociodemographic and medical data [including coronavirus disease 2019 (COVID-19) symptoms and results of RT-PCR tests] were retrieved using questionnaires. The overall seroprevalence was 17.1% [95% confidence interval (CI) 14.9–19.5], with 18.9% (95% CI 15.9–22.2) of the residents and 14.9% (95% CI 11.9–18.4) of the staff having antibodies, which was higher than the seroprevalence in blood donors. The seroprevalence in the 20 NH varied between 0.0% and 45.0%. Fourteen per cent of the staff with antibodies, reported no typical COVID-19 symptoms, while in residents, 51.0% of those with antibodies had no symptoms. The generalised mixed effect model showed a positive association between COVID-19 symptoms and positive serology, but this relation was weaker in residents compared to staff. This study shows that NH are more affected by SARS-CoV-2 than the general population. The large variation between NH, suggests that some risk factors for the spread among residents and staff may be related to the NH. Further, the results suggest that infected people, without the typical COVID-19 symptoms, might play a role in outbreaks.