Development of a foot-and-mouth disease NSP ELISA and its comparison with differential diagnostic methods.

The gene encoding the nonstructural protein (NSP) of O/SKR/2000 foot-and-mouth disease virus (FMDV) was constructed to express under the polyhedron promoter of baculovirus. The expression of NSP was confirmed by indirect immunofluorescence assay (IFA) and Western blotting. The expressed NSP was applied as a diagnostic antigen for indirect-trapping ELISA (I-ELISA). An I-ELISA using monoclonal antibody (Mab) against 3A as trapping antibody was developed to differentiate infected from vaccinated cattle. The diagnostic efficiency of Mab linked I-ELISA was compared and evaluated with baculovirus expressed 3ABC I-ELISA from USDA and Mab (3A) linked E. coli expressed 3ABC I-ELISA from IZSLE through retrospective sero-surveillance. Compared with the two different I-ELISA methods, Mab (3A) linked I-ELISA using baculovirus expressed NSP showed the same level of sensitivity and specificity, indicating that this method is suitable for a differential diagnostic method in cattle.