title: Recombinant nucleocapsid protein based single serum dilution ELISA for the detection of antibodies to infectious bronchitis virus in poultry.
abstract: Avian infectious bronchitis is ubiquitous and highly contagious disease of poultry, with profound effect on commercial poultry production. For effective control of infectious bronchitis virus (IBV), quick and specific diagnosis is of utmost importance. In this study, the virus was isolated from clinical samples from India and the full length nucleocapsid (N) gene was amplified, cloned and expressed in a prokaryotic system. The purified recombinant N protein based single serum dilution enzyme linked immunosorbent assay (ELISA) was developed for IBV to measure specific antibody in the sera of chickens. A total of 310 chicken sera samples were tested using the commercial IDEXX kit along with the assay developed. A linear correlation was obtained between predicted antibody titres at a single working dilution of 1:100 and the corresponding serum titres observed as determined by the standard serial dilution method. Regression analysis was used to construct a standard curve from which an equation was derived which confirmed their correlation. The developed equation was then used to extrapolate predicated ELISA antibody titer from corrected absorbance readings of the single working dilution. The assay proved to be specific (95.8%) and sensitive (96.8%) when compared to the commercial IDEXX ELISA test.
authors.affiliation: Recombinant-DNA Laboratory, Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly 243122, India.
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Pradhan
authors.affiliation: Recombinant-DNA Laboratory, Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly 243122, India.
authors.forename: Nitin M
authors.initials: NM
authors.lastname: Kamble
authors.affiliation: Recombinant-DNA Laboratory, Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly 243122, India.
authors.forename: Aravind S
authors.initials: AS
authors.lastname: Pillai
authors.affiliation: Recombinant-DNA Laboratory, Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly 243122, India.
authors.forename: Satish S
authors.initials: SS
authors.lastname: Gaikwad
authors.affiliation: Recombinant-DNA Laboratory, Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly 243122, India.
authors.forename: Sagar A
authors.initials: SA
authors.lastname: Khulape
authors.affiliation: Project Directorate on Poultry, Hyderabad 500 030, India.
authors.forename: M R
authors.initials: MR
authors.lastname: Reddy
authors.affiliation: Recombinant-DNA Laboratory, Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly 243122, India.
authors.forename: C Madhan
authors.initials: CM
authors.lastname: Mohan
authors.affiliation: Central Avian Research Institute, Izatnagar, Bareilly 243122, India.
authors.forename: Jag Mohan
authors.initials: JM
authors.lastname: Kataria
authors.affiliation: Recombinant-DNA Laboratory, Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, Bareilly 243122, India. Electronic address: sohinimadhan@gmail.com.
authors.forename: Sohini
authors.initials: S
authors.lastname: Dey
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D002645:Chickens; D003001:Cloning, Molecular; D018352:Coronavirus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015870:Gene Expression; D007194:India; D001351:Infectious bronchitis virus / Q000276:immunology*; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000235:genetics; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D011994:Recombinant Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D044967:Serum / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins
keywords: ELISA; Infectious bronchitis virus; Recombinant nucleocapsid protein