Development of monoclonal antibody-based blocking ELISA for detecting SARS-CoV-2 exposure in animals
Development of monoclonal antibody-based blocking ELISA for detecting SARS-CoV-2 exposure in animals
Blog Article
ABSTRACT The global pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) poses a significant threat to public health.Besides humans, SARS-CoV-2 can infect several animal species.Highly sensitive and specific diagnostic reagents and assays are urgently needed for rapid detection and nitrile gloves in a bucket implementation of strategies for prevention and control of the infection in animals.
In this study, we initially developed a panel of monoclonal antibodies (mAbs) against SARS-CoV-2 nucleocapsid protein.To detect SARS-CoV-2 antibodies in a broad spectrum of animal species, an mAb-based blocking enzyme-linked immunosorbent assay (bELISA) was developed.Test validation using a set of animal serum samples with known infection status obtained an optimal percentage of inhibition cut-off value of 17.
6% with diagnostic sensitivity of 97.8% and diagnostic specificity of 98.9%.
The assay demonstrates high repeatability as determined by a low coefficient of variation (7.23%, 4.89%, and 3.
16%) between-runs, within-run, and within-plate, respectively.Testing of samples collected over time from experimentally infected cats showed that the bELISA was able to detect seroconversion as early as 7 days post-infection.Subsequently, the bELISA was applied for testing pet animals with coronavirus disease 2019 (COVID-19)-like symptoms and specific antibody responses were detected in two dogs.
The panel of mAbs generated in this study provides a valuable tool for SARS-CoV-2 diagnostics and research.The mAb-based bELISA provides a serological test in aid of COVID-19 surveillance in animals.IMPORTANCE Antibody tests are commonly used as a diagnostic tool for detecting host immune response following infection.
Serology (antibody) tests complement nucleic acid assays by providing a history of virus exposure, no matter symptoms developed from infection nacrack.com or the infection was asymptomatic.Serology tests for COVID-19 are in high demand, especially when the vaccines become available.They are important to determine the prevalence of the viral infection in a population and identify individuals who have been infected or vaccinated.
ELISA is a simple and practically reliable serological test, which allows high-throughput implementation in surveillance studies.Several COVID-19 ELISA kits are available.However, they are mostly designed for human samples and species-specific secondary antibody is required for indirect ELISA format.
This paper describes the development of an all species applicable monoclonal antibody (mAb)-based blocking ELISA to facilitate the detection and surveillance of COVID-19 in animals.