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Development of a foot-and-mouth disease NSP ELISA and its comparison with differential diagnostic methods
Chang Hee Kweon1, Young Joon Ko, Won I Kim
1Virology Research Division, 480, Anyang, National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry, Republic of Korea. kweonch@mail.nvrqs.go.kr
Abstract:
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.
Insights
Researchers developed a novel indirect-trapping ELISA (I-ELISA) using baculovirus-expressed nonstructural protein (NSP) to differentiate foot-and-mouth disease virus (FMDV) infected from vaccinated cattle.
Area of Science:
- Veterinary Virology
- Immunodiagnostics
- Molecular Biology
Background:
- Foot-and-mouth disease virus (FMDV) poses a significant threat to livestock globally.
- Distinguishing between FMDV infection and vaccination is crucial for effective disease control and trade.
- Current diagnostic methods may require improvement for differential diagnosis.
Purpose of the Study:
- To develop and evaluate a novel diagnostic antigen for FMDV.
- To establish an indirect-trapping ELISA (I-ELISA) capable of differentiating infected from vaccinated animals.
- To assess the diagnostic efficiency of the developed I-ELISA compared to existing methods.
Main Methods:
- The gene encoding the nonstructural protein (NSP) of FMDV was cloned and expressed using a baculovirus system.
- Expression of NSP was confirmed via indirect immunofluorescence assay (IFA) and Western blotting.
- An I-ELISA was developed using monoclonal antibody (Mab) against FMDV 3A protein as the trapping antibody, with baculovirus-expressed NSP as the antigen.
Main Results:
- Baculovirus-expressed FMDV NSP was successfully produced and confirmed.
- The developed Mab (3A) linked I-ELISA demonstrated comparable sensitivity and specificity to other established I-ELISA methods.
- Retrospective sero-surveillance validated the diagnostic performance of the new method.
Conclusions:
- Baculovirus-expressed NSP is a suitable antigen for developing differential diagnostic tools for FMDV.
- The Mab (3A) linked I-ELISA using baculovirus-expressed NSP is an effective method for differentiating FMDV-infected from vaccinated cattle.
- This diagnostic approach can aid in FMDV surveillance and control strategies.