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Detection of antibodies against foot-and-mouth disease virus using a liquid-phase blocking sandwich ELISA (LPBE) with
V K O'Donnell1, D B Boyle, K Sproat
1Instituto de Virologia, Centro de Investigación en Ciencias Veterinarias, INTA, Morón, Provincia de Buenos Aires, Argentina.
Abstract:
A liquid-phase blocking sandwich enzyme-linked immunosorbent assay (ELISA-3D) was developed to detect specific antibodies to the 3D protein in sera from foot-and-mouth disease (FMD) virus (FMDV)-infected animals. The assay uses a nonstructural 3D recombinant protein and two polyclonal antisera, one for capture (bovine) and the other for detector (guinea pig). The specificity of the assay was demonstrated by negative results with 101 sera of cattle from the FMD-free zone in Argentina and with bovine and porcine sera raised against various RNA and DNA viruses. The ELISA-3D was able to detect antibodies in cattle after natural or experimental infection with FMDV of A, O, or C types as early as 5 days postinfection and at later stages in persistently infected animals. Comparison of the results with those obtained with the routinely used agar gel immunodiffusion test and a previously described ELISA, both employing a partially purified virus-infection-associated antigen, shows that the ELISA-3D is highly sensitive and specific and gives reproducible results. Its use as a tool for monitoring viral activity and for certification of FMDV-free animals is recommended.
Insights
A new enzyme-linked immunosorbent assay (ELISA-3D) accurately detects foot-and-mouth disease virus (FMDV) antibodies in cattle. This highly sensitive and specific assay aids in monitoring viral activity and certifying FMDV-free animals.
Area of Science:
- Veterinary Virology
- Immunodiagnostics
- Animal Health
Background:
- Foot-and-mouth disease virus (FMDV) poses a significant threat to livestock globally.
- Accurate diagnostic tools are crucial for FMDV surveillance and control.
- Existing diagnostic methods may have limitations in sensitivity or specificity.
Purpose of the Study:
- To develop and validate a novel liquid-phase blocking sandwich enzyme-linked immunosorbent assay (ELISA-3D).
- To detect specific antibodies against the FMDV 3D protein in infected animals.
- To evaluate the assay's performance compared to established diagnostic techniques.
Main Methods:
- Development of a sandwich ELISA utilizing a recombinant FMDV 3D protein.
- Inclusion of bovine and guinea pig polyclonal antisera for antibody capture and detection.
- Validation of specificity using sera from FMDV-free cattle and animals infected with other viruses.
- Assessment of sensitivity in experimentally and naturally infected cattle.
Main Results:
- The ELISA-3D demonstrated high specificity, yielding negative results with FMDV-free cattle sera and sera from animals infected with other viruses.
- Antibodies were detected as early as 5 days post-infection in cattle infected with FMDV types A, O, and C.
- The assay successfully identified antibodies in persistently infected animals.
- ELISA-3D showed superior sensitivity and specificity compared to agar gel immunodiffusion and a previously described ELISA.
Conclusions:
- The developed ELISA-3D is a highly sensitive, specific, and reproducible diagnostic tool for FMDV antibody detection.
- This assay is suitable for monitoring FMDV circulation in animal populations.
- ELISA-3D can be effectively used for the certification of FMDV-free animals and herds.