The validity of blood spotted on to filter-paper (BSOFP) eluates for the detection of antibodies against hepatitis A virus (HAV) was investigated in 718 individuals (children and adults) during a field study in a small area in Rio de Janeiro State, Brazil. Serum samples were considered the 'gold standard'. BSOFP eluates were analyzed by 2 different techniques: microplate competitive enzyme-linked immunosorbent assay (ELISA) of the whole study group and microparticle enzyme immune assay (MEIA) of a subsample of 59 individuals. For BSOFP eluates by ELISA, sensitivity and specificity were 89.6% (95% CI: 84.7-93.1) and 97.5% (95% CI: 95.6-98.7), respectively. For a seroprevalence of anti-HAV antibodies of 32%, the positive predictive value was 94.5% (95% CI: 90.3-97.0) and the negative predictive value was 95.2% (95% CI: 92.8-96.8). The test efficiency was 95.0% (95% CI: 93.1-96.4). Similar results were found for BSOFP eluates by MEIA. Agreement between the 2 techniques used for BSOFP (ELISA and MEIA) was also high (kappa = 0.93). These results encourage the more widespread application of BSOFP as a means of surveillance for large-scale epidemiological studies for hepatitis A.