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Formation of subviral particles of the capsid protein VP2 of infectious bursal disease virus and its application in serological diagnosis
- Source :
- Journal of virological methods. 157(1)
- Publication Year :
- 2008
-
Abstract
- Infectious bursal disease virus (IBDV) is an immunosuppressive disease of young chicken characterized by severe depletion of B-lymphocytes in the bursa of Fabricius. To provide antigen for diagnostic tests, its major structural protein VP2 was expressed in the yeast Saccharomyces cerevisiae. Electron microscopy of purified VP2 protein demonstrated that when expressed from yeast cells VP2 protein forms subviral particles (SVPs) of approximately 20 nm in diameter. A recombinant VP2 antigen-based single serum dilution enzyme linked immunosorbent assay (ELISA) using the SVPs detected IBDV specific antibodies in chickens. A linear relationship was found between the predicted antibody titres at a single working dilution of 1:1000 and the corresponding observed serum titres when 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 equation was then used to convert the corrected absorbance readings of the single working dilution directly into the predicted ELISA antibody titres. The assay proved to be sensitive, specific and accurate as compared to the serum neutralization test and agar gel immunodiffusion test. The recombinant VP2 antigen is a suitable alternative to whole viral antigen.
- Subjects :
- animal structures
Birnaviridae
Serial dilution
Virosomes
Gene Expression
Enzyme-Linked Immunosorbent Assay
Chick Embryo
Saccharomyces cerevisiae
Biology
Antibodies, Viral
Sensitivity and Specificity
Virus
Infectious bursal disease
Antigen
Virology
medicine
Animals
Bursa of Fabricius
Cloning, Molecular
Viral Structural Proteins
medicine.disease
Ouchterlony double immunodiffusion
biology.organism_classification
Birnaviridae Infections
Recombinant Proteins
Microscopy, Electron
biology.protein
Antibody
Chickens
Subjects
Details
- ISSN :
- 01660934
- Volume :
- 157
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Journal of virological methods
- Accession number :
- edsair.doi.dedup.....0587a121c41ece97c0fe3ba252612adf