title: Evaluation of a nucleoprotein-based enzyme-linked immunosorbent assay for the detection of antibodies against infectious bronchitis virus.
abstract: As an immunogen of the coronavirus, the nucleoprotein (N) is a potential antigen for the serological monitoring of infectious bronchitis virus (IBV). In this report, recombinant N protein from the Beaudette strain of IBV was produced and purified from Escherichia coli as well as Sf9 (insect) cells, and used for the coating of enzyme-linked immunosorbent assay (ELISA) plates. The N protein produced in Sf9 cells was phosphorylated whereas N protein from E. coli was not. Our data indicated that N protein purified from E. coli was more sensitive to anti-IBV serum than the protein from Sf9 cells. The recombinant N protein did not react with the antisera to other avian pathogens, implying that it was specific in the recognition of IBV antibodies. In addition, the data from the detection of field samples and IBV strains indicated that using the recombinant protein as coating antigen could achieve an equivalent performance to an ELISA kit based on infected material extracts as a source of antigen(s). ELISAs based on recombinant proteins are safe (no live virus), clean (only virus antigens are present), specific (single proteins can be used) and rapid (to respond to new viral strains and strains that cannot necessarily be easily cultured).
authors.affiliation: School of Animal and Microbial Sciences, University of Reading, UK.
authors.forename: Hongying
authors.initials: H
authors.lastname: Chen
authors.forename: Bruce
authors.initials: B
authors.lastname: Coote
authors.forename: Simon
authors.initials: S
authors.lastname: Attree
authors.forename: Julian A
authors.initials: JA
authors.lastname: Hiscox
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000302:isolation & purification*; D000956:Antigens, Viral / Q000276:immunology; D002645:Chickens; D018352:Coronavirus Infections / Q000175:diagnosis / Q000662:veterinary; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D004926:Escherichia coli / Q000235:genetics; D006168:Guinea Pigs; D001351:Infectious bronchitis virus / Q000276:immunology* / Q000302:isolation & purification; D009698:Nucleoproteins / Q000145:classification / Q000276:immunology*; D011201:Poultry Diseases / Q000175:diagnosis; D011994:Recombinant Proteins / Q000276:immunology; D012449:Safety; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms; D013997:Time Factors
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D009698:Nucleoproteins; D011994:Recombinant Proteins