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Recombinant nucleocapsid protein based single serum dilution ELISA for the detection of antibodies to infectious bronchitis virus in poultry.
- Source :
-
Journal of virological methods [J Virol Methods] 2014 Dec; Vol. 209, pp. 1-6. Date of Electronic Publication: 2014 Aug 27. - Publication Year :
- 2014
-
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.<br /> (Copyright © 2014 Elsevier B.V. All rights reserved.)
- Subjects :
- Animals
Chickens
Cloning, Molecular
Coronavirus Infections diagnosis
Coronavirus Infections virology
Enzyme-Linked Immunosorbent Assay methods
Gene Expression
India
Molecular Sequence Data
Poultry Diseases virology
Recombinant Proteins genetics
Sensitivity and Specificity
Sequence Analysis, DNA
Serum immunology
Antibodies, Viral blood
Coronavirus Infections veterinary
Infectious bronchitis virus immunology
Nucleocapsid Proteins genetics
Poultry Diseases diagnosis
Subjects
Details
- Language :
- English
- ISSN :
- 1879-0984
- Volume :
- 209
- Database :
- MEDLINE
- Journal :
- Journal of virological methods
- Publication Type :
- Academic Journal
- Accession number :
- 25173423
- Full Text :
- https://doi.org/10.1016/j.jviromet.2014.08.015