Development of a competitive enzyme-linked immunosorbent assay for detection of turkey coronavirus antibodies
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Published on: September 6, 2016
James S Guy1, Lynda G Smith, Jamie J Breslin
1Department of Microbiology, Pathology, and Parasitology, College of Veterinary Medicine, North Carolina State University, Raleigh 27606, USA.
Insights
A new competitive enzyme-linked immunosorbent assay (cELISA) accurately detects turkey coronavirus (TCV) antibodies. This rapid, sensitive, and specific test aids in diagnosing TCV infections in turkeys.
Area of Science:
- Veterinary Immunology
- Virology
- Diagnostic Assays
Background:
- Turkey coronavirus (TCV) poses a significant threat to poultry health.
- Accurate and reliable diagnostic tools are crucial for controlling TCV outbreaks.
- Existing serological methods may have limitations in sensitivity or specificity.
Purpose of the Study:
- To develop and validate a competitive enzyme-linked immunosorbent assay (cELISA) for detecting TCV antibodies.
- To evaluate the sensitivity and specificity of the developed cELISA compared to established methods.
Main Methods:
- A cELISA was designed using recombinant TCV nucleocapsid (N) protein and a specific monoclonal antibody.
- The assay's performance was assessed using 1269 reference, experimental, and field-origin turkey sera.
- Comparison was made against the indirect fluorescent antibody test (IFAT), with discordant results further analyzed by western immunoblot.
Main Results:
- The cELISA demonstrated high concordance with IFAT, showing 92.9% sensitivity and 96.2% specificity.
- The assay specifically detected antibodies against TCV and infectious bronchitis virus but not other avian viruses.
- Western immunoblot analysis corroborated the cELISA's specificity.
Conclusions:
- The developed cELISA is a rapid, sensitive, and specific serological tool for TCV antibody detection in turkeys.
- This assay offers a reliable method for diagnosing TCV infections and monitoring flock immunity.
- The cELISA can aid in the effective management and control of TCV in poultry populations.
Abstract:
A competitive enzyme-linked immunosorbent assay (cELISA) was developed for detection of turkey coronavirus (TCV) antibodies. The cELISA utilized a recombinant baculovirus (Autographa californica nuclear polyhedrosis virus)-expressed TCV nucleocapsid (N) protein and biotin-labeled TCV N protein-specific monoclonal antibody. Sensitivity and specificity of the cELISA for detection of TCV antibodies were determined by comparison with the indirect fluorescent antibody test (IFAT) with 1269 reference, experimentally derived, and field-origin sera. Sera with discordant cELISA and IFAT results were further evaluated by western immunoblot analyses. The cELISA detected antibodies specific for TCV and infectious bronchitis virus, a closely related coronavirus, but did not detect antibodies specific for other avian viruses. A high degree of concordance was observed between the cELISA and IFAT; sensitivity and specificity of the cELISA relative to IFAT were 92.9% and 96.2%, respectively. Western immunoblot analyses provided additional evidence of cELISA specificity. The findings indicate that the cELISA is a rapid, sensitive, and specific serologic test for detection of TCV antibodies in turkeys.
