A multiplex enzyme-linked immunosorbent assay (ELISA) to detect antibodies to Bovine viral diarrhea virus (BVDV), Bovine herpesvirus 1 (BHV-1), Bovine parainfluenza virus 3 (BPV-3), and Bovine respiratory syncytial virus (BRSV) was developed on the commercially available Meso Scale Discovery (MSD) platform. The multiplex ELISA used the same antigens (relatively crude lysates of cultured virus) that are used in the current in-house separate ELISAs. Testing a panel of field samples demonstrated the multiplex assay could simultaneously detect the presence of several antibodies in a single assay and that there was good agreement between the multiplex ELISA and the in-house ELISAs. The use of a multiplex ELISA could offer significant benefits for routine serological testing through reduced staff time, reduced reagent cost, and streamlined laboratory operation.