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Indirect ELISA using recombinant nonstructural protein 3D to detect foot and mouth disease virus infection associated
Sonalika Mahajan1, Jajati Keshari Mohapatra1, Laxmi Kant Pandey1
1Indian Council of Agricultural Research, IVRI Campus, Mukteswar, Uttarakhand 263138, India.
Abstract:
Foot-and-mouth disease (FMD) is a highly infectious disease of transboundary importance. Routine biannual vaccination along with surveillance activities is seen as the principal to control FMD in India. Non-structural protein (NSP) based immunoassays are the test of choice for the differentiation between infected and vaccinated population. In this study, 3D protein of FMD virus was expressed in Escherichia coli and an indirect ELISA (I-ELISA) was developed to detect 3D-antibodies in the infected bovines. 3D I-ELISA demonstrated comparable diagnostic sensitivity (97.6%) but lower specificity (80.8%) as compared to the in-house r3AB3 I-ELISA. However, the specificity values varied significantly for naïve and vaccinated samples and were observed to be 98.42% and 76.93%, respectively. A moderate degree of concordance (88.5%) was observed between the overall results of two ELISAs. 3D I-ELISA displayed a considerably lower specificity in uninfected vaccinated samples, thereby suggesting against its application for serosurveillance in intensively vaccinated population. However by virtue of its high diagnostic sensitivity and longer duration of persistence of 3D-antibody post-infection, 3D I-ELISA could be adopted for seroepidemiological investigations in regions not practicing vaccination and could be extended to susceptible species which are generally not covered by vaccination.
Insights
A new indirect ELISA (I-ELISA) using the 3D protein of Foot-and-mouth disease virus (FMDV) shows high sensitivity for detecting infected bovines. However, its lower specificity in vaccinated animals limits its use in routine surveillance programs.
Area of Science:
- Veterinary Virology
- Immunodiagnostics
- Disease Surveillance
Background:
- Foot-and-mouth disease (FMD) is a highly contagious transboundary animal disease.
- Control strategies in India rely on vaccination and surveillance.
- Differentiating infected from vaccinated animals (DIVA) is crucial for effective FMD management.
Purpose of the Study:
- To develop and evaluate an indirect ELISA (I-ELISA) for detecting antibodies against the FMD virus 3D protein.
- To assess the diagnostic performance of the 3D I-ELISA in differentiating infected from vaccinated bovines.
Main Methods:
- The FMD virus 3D protein was expressed in Escherichia coli.
- An indirect ELISA (I-ELISA) was developed using the recombinant 3D protein.
- Diagnostic sensitivity and specificity were determined by comparing results with an in-house r3AB3 I-ELISA and known infected/vaccinated samples.
Main Results:
- The 3D I-ELISA demonstrated high diagnostic sensitivity (97.6%).
- Overall specificity was 80.8%, with significant variations: 98.42% for naïve and 76.93% for vaccinated samples.
- A moderate concordance (88.5%) was observed between the 3D I-ELISA and the r3AB3 I-ELISA.
Conclusions:
- The 3D I-ELISA shows promise for seroepidemiological investigations in non-vaccinated regions due to its high sensitivity and persistent antibody detection.
- Its lower specificity in vaccinated populations makes it unsuitable for routine serosurveillance in intensively vaccinated areas.
- The assay could be extended to non-bovine susceptible species not covered by vaccination programs.
