Since the discovery of the Norwalk virus (NV) by immune electron microscopy (IEM) in 1972, serologic studies with this virus have relied on particle-positive fecal material from infected volunteers as the source of antigen because it has not been possible to propagate this virus in cell culture. However, the recent cloning of the W (strain 8FIIa) genome and expression of the capsid protein in a baculovirus system to form ''virus-like particles'' has provided a consistent source of antigen (designated rNV). The purpose of the present study was to compare the antigenicities of these rW particles with those of native W antigen derived from human fecal material by using well-characterized sera obtained from earlier studies. In IEM studies, the rNV antigen reacted with NV-specific antibodies in a manner similar to that observed previously when particle-positive fecal material was used as antigen. In addition, a direct enzyme-linked immunosorbent assay, in which the rW antigen was used as antigen, proved efficient and specific for the detection of serologic responses to W compared with the previously established techniques of IEM and blocking antibody immunoassays in which particle-positive fecal material was used as the antigen. The availability of an unlimited source of antigen will enable serologic studies that will greatly increase our understanding of the epidemiology of W and its role in human enteric illness.