Development and Validation of Serotype-Specific Blocking ELISA for the Detection of Anti-FMDV O/A/Asia1/SAT2 Antibodies

Viruses. 2024 Sep 10;16(9):1438. doi: 10.3390/v16091438.

Abstract

Foot-and-mouth disease (FMD) is one of the most infectious viral transboundary diseases of livestock, which causes devastating global economic losses. Different enzyme-linked immunosorbent assays (ELISAs) are used for sero-surveillance of the foot-and-mouth disease virus (FMDV). However, more sensitive, accurate, and convenient ELISAs are still required to detect antibodies against FMDV serotypes. The primary goal of this study was to establish serotype-specific monoclonal antibody (mAb)-based blocking ELISAs (mAb-bELISAs) that would provide better performance characteristics or be equivalent in performance characteristics compared with a conventional polyclonal antibody (pAb)-based competitive ELISA (pAb-cELISA). Four mAb-bELISAs were developed using FMDV serotype-specific mAbs for the detection of anti-FMDV/O/A/Asia1/SAT2 antibodies. Using a 50% cut-off, all four mAb-bELISAs exhibited species-independent 99.74%, 98.01%, 96.59%, and 98.55% diagnostic specificity (DSp) and 98.93%, 98.25%, 100%, and 87.50% diagnostic sensitivity (DSe) for FMDV serotypes O, A, Asia1, and SAT2, respectively. In addition, a 100% DSe of serotypes O- and SAT2-specific mAb-bELISAs was observed for porcine sera when the cut-off was 30%. All mAb-bELISAs developed in this study displayed high repeatability/reproducibility without cross-reactivity. Finally, the diagnostic performance of mAb-bELISAs was found to be better than or equivalent to compared with pAb-cELISAs, suggesting that mAb-bELISAs can be used to replace existing pAb-ELISAs for the detection of antibodies against these four FMDV serotypes.

Keywords: DSe; DSp; FMDV serotypes; bELISA; monoclonal antibody; polyclonal antibodies.

Publication types

  • Validation Study

MeSH terms

  • Animals
  • Antibodies, Monoclonal* / immunology
  • Antibodies, Viral* / blood
  • Antibodies, Viral* / immunology
  • Cattle
  • Enzyme-Linked Immunosorbent Assay* / methods
  • Foot-and-Mouth Disease Virus* / classification
  • Foot-and-Mouth Disease Virus* / immunology
  • Foot-and-Mouth Disease* / diagnosis
  • Foot-and-Mouth Disease* / immunology
  • Foot-and-Mouth Disease* / virology
  • Mice
  • Reproducibility of Results
  • Sensitivity and Specificity*
  • Serogroup*
  • Swine
  • Swine Diseases / diagnosis
  • Swine Diseases / immunology
  • Swine Diseases / virology

Substances

  • Antibodies, Viral
  • Antibodies, Monoclonal

Grants and funding

This research received no funding.