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Development of A recombinant nucleocapsid based indirect ELISA for the detection of antibodies to avian metapneumovirus subtypes, A, B, and C
- Source :
- Veterinary immunology and immunopathology. 231
- Publication Year :
- 2020
-
Abstract
- Nucleocapsid (N) protein is the most highly expressed of all avian metapneumovirus (aMPV) viral proteins and stimulates a substantial immune response in infected animals. Codon optimized recombinant N (rec-N) protein from aMPV subtypes A, B, and C were expressed using the baculoviral expression system in Trichoplusia ni (Tni) insect cells. A mixture of purified rec-N antigens from each subtype was used as a coating antigen and was evaluated in indirect ELISA (iELISA) to assess antibody response in serum samples collected from experimentally infected chickens and turkeys with different aMPV subtypes. Also, archived field serum samples that were collected from different poultry submissions were used. Receiver operating characteristic (ROC) analysis was performed using chicken and turkey serum samples that were confirmed by indirect fluorescent antibody (IFA) test for serostatus (positive n = 270, negative n = 610). The ROC analysis showed sensitivity and specificity of 97 % at a cut-off value of 0.25. The rec-N iELISA was compared with a commercial whole virus-based APV kit. The rec-N iELISA showed comparable results in detecting antibody response in aMPV infected chicken sera but was more sensitive in detecting early antibody response in aMPV infected turkey serum samples. Our results further confirm the presence of aMPV antibodies in Canadian domestic poultry populations. The developed aMPV-rec N iELISA offers a safe and valuable alternative to whole virus-based iELISA for serodiagnosis and seroepidemiological surveillance of the disease in domestic poultry.
- Subjects :
- Indirect elisa
Turkeys
040301 veterinary sciences
Immunology
Enzyme-Linked Immunosorbent Assay
Antibodies, Viral
Virus
law.invention
0403 veterinary science
03 medical and health sciences
Immune system
Antigen
law
Chlorocebus aethiops
Trichoplusia
Animals
Metapneumovirus
Nucleocapsid
Vero Cells
Poultry Diseases
030304 developmental biology
0303 health sciences
Paramyxoviridae Infections
General Veterinary
biology
04 agricultural and veterinary sciences
biology.organism_classification
Virology
biology.protein
Recombinant DNA
Antibody
Chickens
Subjects
Details
- ISSN :
- 18732534
- Volume :
- 231
- Database :
- OpenAIRE
- Journal :
- Veterinary immunology and immunopathology
- Accession number :
- edsair.doi.dedup.....f035054bcd40ead44fe4eaa2fc990a20