Key result
A novel blocking ELISA targeting the conserved 'AEKNPLE' epitope in FMDV NSP 3A successfully differentiated infected from vaccinated animals with specificities of 96.6-100% and sensitivities of 61.4-93.2%.
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The developed 3A-Mab-bELISA can serve as an effective companion diagnostic test for the 3A epitope-deleted FMDV marker vaccine to differentiate infected from vaccinated animals.
Foot-and-mouth disease (FMD) is a devastating animal disease. Strategies for differentiation of infected from vaccinated animals (DIVA) remain very important for controlling disease. Development of an epitope-deleted marker vaccine and accompanying diagnostic method will improve the efficiency of DIVA. Here, a monoclonal antibody (Mab) was found to recognize a conserved "AEKNPLE" epitope spanning amino acids 109-115 of non-structural protein (NSP) 3A of foot-and-mouth disease virus (FMDV; O/Tibet/CHA/99 strain), which could be deleted by a reverse-genetic procedure. In addition, a blocking ELISA was developed based on this Mab against NSP 3A, which could serve as a matching test for a negative-marker vaccine. The criterion of this blocking ELISA was determined by detecting panels of sera from different origins. The serum samples with a percentage inhibition (PI) equal or greater than 50% were considered to be from infected animals, and those with <50% PI were considered to be from non-infected animals. This test showed similar performance when compared with other 2 blocking ELISAs based on an anti-NSP 3B Mab. This is the first report of the DIVA test for an NSP antibody based on an Mab against the conserved and predominant "AEKNPLE" epitope in NSP 3A of FMDV.
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Fu et al. (2017) studied Foot-and-mouth disease (FMD) (n=1,989). 3A-Mab-bELISA vs. 3B-Mab-bELISA and PrioCHECK FMDV NS ELISA was evaluated on Diagnostic sensitivity and specificity at a 50% percentage inhibition (PI) cutoff. A novel blocking ELISA targeting the conserved 'AEKNPLE' epitope in FMDV NSP 3A successfully differentiated infected from vaccinated animals with specificities of 96.6-100% and sensitivities of 61.4-93.2%.
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