Foot-and-mouth disease (FMD) is a highly infectious disease of cloven-hoofed animals with a significant economic impact. Early diagnosis and effective prevention and control could reduce the spread of the disease which could possibly minimize economic losses. Epitope characterization based on monoclonal antibodies provide essential information for developing diagnostic assays and vaccine designs. In this study, monoclonal antibodies raised against FMD virus (FMDV) were produced. Sixty-six monoclonal antibodies demonstrated strong reactivity and specificity to FMDV. The purified monoclonal antibodies were further used for bio-panning to select phage expressing specific epitopes from phage-displayed 12 mer-peptide library. The phage peptide sequences were analyzed using multiple sequence alignment and evaluated by peptide ELISA. Two hybridoma clones secreted monoclonal antibodies recognizing linear epitopes on VP2 of FMDV serotype O. The non-neutralizing monoclonal antibody 6F4.D11.B6 recognized the residues 67–78 on antigenic site 2 resinding in VP2, while the neutralizing monoclonal antibody 8D6.B9.C3 recognized a novel linear epitope encompassing residues 115–126 on VP2. This information and the FMDV-specific monoclonal antibodies provide valuable sources for further study and application in diagnosis, therapeutics and vaccine designs to strengthen the disease prevention and control measures.

Foot-and-mouth disease (FMD) is one of the most infectious viral diseases of coven-hoofed animals. The crucial strategy for FMD control is the combination of animal quarantine and rapid onsite diagnostic assay capable of differential infected from vaccinated animals (DIVA). Herein, we have developed a lateral flow immunoassay to detect specific antibodies against FMDV non-structural protein (NSP), based on recombinant mu3ABC produced by E. coli as an antibody detector, so called mu3ABC strip test. Performance of mu3ABC strip test was examined using 566 field serum samples from cattle, pigs, and goats and compared with the commercial ELISA kit, resulting in the diagnostic sensitivity (DSn) and specificity (DSp) of 88.18% and 92.56%, respectively. Additionally, efficacies of the mu3ABC strip test and commercial ELISA were determined using 200 reference bovine sera (100 positives, 100 negatives) classified by the Regional Reference Laboratory for Foot and Mouth Disease in South East Asia (RRL). The result reveals that DSn and DSp of mu3ABC strip test were 78% and of 91%, respectively while the commercial ELISA kit had DSn of 69%and DSp of 100%. The developed mu3ABC strip tests are effective and can be used to differentiate the infected animals in parallel with the sero-surveillance ELISA test.

Foot-and-mouth disease (FMD) is one of the most infectious viral diseases of cloven-hoofed animals. The crucial strategy for FMD control is the combination of animal quarantine and a rapid onsite diagnostic assay capable of differentiation of infected from vaccinated animals (DIVA). Herein, we have developed a lateral flow immunoassay to detect specific antibodies against FMDV non-structural protein (NSP), based on recombinant mu3ABC produced by E. coli as an antibody detector, the so-called mu3ABC strip test. The performance of this test was examined using 366 field serum samples from cattle, pigs, and goats and compared with the commercial ELISA kit, resulting in a diagnostic sensitivity (DSn) and specificity (DSp) of 98.05% and 93.16%, respectively. Additionally, efficacies of this test and commercial ELISA were determined using 200 reference bovine serum samples (100 positives, 100 negatives) classified by the Regional Reference Laboratory for Foot-and-Mouth Disease in South East Asia (RRL). The result reveals that the DSn and DSp of the test were 78% and 91%, respectively, while those for the commercial ELISA kit were 69% and 100%, respectively. The developed mu3ABC strip tests are effective and can be used to differentiate the infected animals in parallel with the sero-surveillance ELISA test.