The Kallestad Pathfinder enzyme immunoassay (EIA) for the rapid detection of respiratory syncytial virus (RSV) antigen was compared with virus culture and direct fluorescent antibody (DFA) to determine the reliability of the EIA. During two consecutive winter respiratory seasons, 270 nasopharyngeal wash specimens were tested. RSV was detected in culture by the presence of cytopathic effect and/or an indirect immunofluorescence assay. The sensitivity of the Pathfinder EIA in comparison with isolation in tube culture was 72% (73 of 101) and the specificity was 99% (167 of 169). During the second year of the evaluation period, DFA was performed on all specimens. The sensitivity of the DFA compared with isolation in tube culture was 94%. This study indicates that the Pathfinder EIA is a very specific test for diagnosis of RSV infections, but lacks sensitivity in comparison with tube culture or direct immunofluorescence.