A trapping enzyme-linked immunosorbent assay (ELISA) has been evaluated for the differentiation of foot-and-mouth disease virus (FMDV) strains using a panel of seven anti-serotype O monoclonal antibodies (MAbs). The variation of results within and between tests performed on the same day and on different days was examined using three strains of FMDV. Criteria for establishing antigenic differences between the strains as defined by the individual MAbs are proposed based on the variability measured, which can be used as standards by workers performing this test with other MAbs and FMDV strains.