A new, simple enzyme-linked immunosorbent assay (ELISA) is described which is performed directly on infected and fixed cell cultures in microtitre plates. It permits large scale screening of antiviral monoclonal antibodies and differentiation of specific antibodies from those usually responsible for high background reactions in other ELISA techniques. Time consuming purification of antigens is thus avoided. The plaque/focus immunoassay is also applicable to titration of antibodies in patients' sera and antigens in lytically or non-lytically virus-infected cells. It may also be used to localize antigens in different cell compartments. This immunoassay requires no special equipment and results may be evaluated either with the naked eye or using a light microscope.