HOME JOURNALS CONTACT

International Journal of Poultry Science

Year: 2012 | Volume: 11 | Issue: 5 | Page No.: 349-360
DOI: 10.3923/ijps.2012.349.360
Confirmation of B-Cell Epitope of Nucleoprotein of Newcastle Disease Viruses by Dot-ELISA and Peptide ELISA
R.N. Ramani Pushpa, J. John Kirubaharan and A. Koteeswaran

Abstract: A conserved B-cell epitope of nucleoprotein with the amino acid sequence “FLDLMRA” was found to be located in the conserved region between 443GETQFLDLMRAVANS457 of nucleoprotein of all four vaccine viruses used in South India. The conserved region incorporating the epitope 443GETQFLDLMRAVANS457 has been synthesized as a custom peptide. The immunodominance of the epitope was confirmed by dot-ELISA and peptide ELISA. Peptide ELISA developed using the entire conserved region was found to produce a consistent result with HI and Whole Virus Protein (WVP) ELISA. The immunodominance and immunogenicity was identified and confirmed by significant positive correlation (p<0.01) between peptide ELISA with HI and with WVP ELISA and also by Probability Plot (PP). Based on PP it could be appreciated that all points cluttered towards the expected line confirming the closeness of the results which is further strengthened by observation from ROC curves. The empirical ROC values for HI Vs Peptide ELISA and WVP ELISA VS Peptide ELISA were 0.97 and 1.00 respectively which are more than the standard value 0.8 confirming the closeness of the results proving immunodominance of the epitope and utility of the epitope to serve as a marker in DIVA vaccine.

Fulltext PDF

How to cite this article
R.N. Ramani Pushpa, J. John Kirubaharan and A. Koteeswaran, 2012. Confirmation of B-Cell Epitope of Nucleoprotein of Newcastle Disease Viruses by Dot-ELISA and Peptide ELISA. International Journal of Poultry Science, 11: 349-360.

Keywords: Newcastle disease virus, NP gene, B cell epitopes, immunodominance, HI- ELISA and marker

REFERENCES
  • Aldous, E.W. and D.J. Alexander, 2001. Detection and differentiation of Newcastle disease virus (avian paramyxovirus type 1). Avian Pathol., 30: 117-128.
    CrossRef    PubMed    

  • Alexander, D.J., 1997. Newcastle Disease and other Avain Paramyxoviridae Infections. In: Diseases of Poultry, Calnek, B.W., H.J. Barnes, C.W. Beard, L.R. McDougald and Y.M. Saif (Eds.). 10th Edn., Iowa State University Press, Ames, pp: 541-570

  • Alexander, D.J., 2001. Newcastle disease. Br. J. Poult. Sci., 42: 5-22.
    CrossRef    Direct Link    

  • Alexander, D.J., 2003. Newcastle Disease, Other Paramyxo Viruses and Pneumovirus Infections. In: Diseases of Poultry, Saif, Y.M., H.J. Barnes, J.R. Ghison, A.M. Fadly, Mc Douglad and D.E. Sweyne (Eds.). 11th Edn., Iowa State Press, Amcs IA, pp: 63-100

  • Babiuk, L.A., 1999. Broadening the approaches to developing more effective vaccines. Vaccine, 17: 1587-1595.
    CrossRef    Direct Link    

  • Bates, P.A. and M.J. Sternberg, 1999. Model building by comparison at CASP3: Using expert knowledge and computer automation. Proteins, 3: 47-54.
    Direct Link    

  • Bates, P.A., L.A. Kelley, R.M. MacCallum and M.J.E. Sternberg, 2001. Enhancement of protein modeling by human intervention in applying the automatic programs 3D-JIGSAW and 3D-PSSM. Proteins, 5: 39-46.
    CrossRef    PubMed    

  • Contreras-Moreira, B. and P.A. Bates, 2002. Domain fishing: A first step in protein comparative modelling. Bioinformatics, 18: 1141-1142.
    Direct Link    

  • Dechamma, H.J., V. Dighe, C. Ashok Kumar, R.P. Singh, M. Jagadish and Satish Kumar, 2006. Identification of T-helper and linear B epitope in the hypervariable region of nucleocapsid protein of PPRV and its use in the development of specific antibodies to detect viral antigen. Vet. Microbiol., 118: 201-211.
    CrossRef    Direct Link    

  • Errington, W., M. Steward and P.T. Emmerson, 1995. A diagnostic immunoassay for Newcastle disease virus based on the nucleocapsid protein expressed by a recombinant baculovirus. J. Virol. Methods, 55: 357-365.
    CrossRef    Direct Link    

  • Hemmatzadeh, F. and A. Zadeh, 2006. A specific dot-blot immuno assay for Newcastle disease serodiganosis. Int. J. Poult. Sci., 5: 1079-1081.

  • Folitse, R., D.A. Halvonson and V. Sivanandan, 1999. A dot immuno assay (dot blot- ELISA) for early detection of Newcastle disease antibodies in chickens. Avian Dis., 42: 14-19.

  • Fooks, A.R., J.R. Stephenson, A. Warnes, A.B. Dowsett, B.K. Rima and G.W.G. Wilkinson, 1993. Measles virus nucleocapsid protein expressed in insect cells assembles into nucleocapsid-like structures. J. Gen. Virol., 74: 1439-1444.
    Direct Link    

  • Hummel, K.B., D.D. Erdman, J. Heath and W.J. Bellini, 1992. Baculovirus expression of the nucleoprotein gene of measles virus and utility of the recombinant protein in diagnostic enzyme immunoassays. J. Chin. Phicobiol., 30: 2874-2880.
    Direct Link    

  • Jacobson, R.H., 1998. Validation of serological assays for diagnosis of infectious diseases. Revue Sci. Technique (Int. Office Epizootics), 17: 469-526.
    Direct Link    

  • John Kirubaharan, J., R. Velmurugan, K. Kumanan and A. Albert, 2000. Inactivated vaccines against Newcastle diseases virus 2 Humoral immune response. Int. Vet. J., 77: 563-563.

  • Kho, C.L., W.S. Tan, B.T. Tey and K. Yusoff, 2003. Newcastle disease virus nucleocapsid protein: Self assembly and length determination domains. J. Gen. Virol., 84: 2163-2168.
    Direct Link    

  • Linde, G.A., M. Granstrom and C. Orvell, 1987. Immunoglobulin class and immunoglobulin G subclass enzyme-linked immunosorbent assays compared with microneutralization assay for serodiagnosis of mumps infection and determination of immunity. J. Clin. Microbiol., 25: 1653-1658.
    Direct Link    

  • Makkay, A.M., P.J. Krell and E. Nagy, 1999. Antibody detection based differential ELISA for NDV-infected or vaccinated chickens versus NDV HN subunit vaccinated chickens. Vet. Microbiol., 66: 209-222.
    Direct Link    

  • Manoharan, S., M. Parthiban, T.G. Prabhakar, G. Ravikumar, A. Koteeswaran, N.D.J. Chandran and G. Rajavelu, 2004. Rapid serological profiling by an Immunocomb-based Dot-enzyme-linked immunosorbent test for three major poultry diseases. Vet. Res. Commun., 28: 339-346.
    CrossRef    Direct Link    

  • Mebatsion, T., M.J.M. Koolen, L.T.C. de Vaan, N. de Haas and M. Braber et al., 2002. Newcastle Disease Virus (NDV) marker vaccine: An immunodominant epitope on the nucleoprotein gene of NDV can be deleted or replaced by a foreign epitope. J. Virol., 76: 10138-10146.
    CrossRef    Direct Link    

  • Reynolds, D.L. and A.D. Maraqa, 2000. Protective immunity against Newcastle disease: The role of cell mediated immunity. Avian Dis., 44: 145-154.

  • De MacArio, E.C., J.D. Folds, R.M. Nakamura and M.N. Robert, 1997. Manual of Clinical Laboratory Immunology. 5th Edn., Amer Society for Microbiology, Washington, DC., Pages: 1255

  • Seal, B.S., D.J. King and J.D. Bennett, 1995. Characterization of Newcastle disease virus isolates by reverse transcription PCR coupled to direct nucleotide sequencing and development of sequence database for pathotype prediction and molecular epidemiological analysis. J. Clin. Microbiol., 33: 2624-2630.
    Direct Link    

  • Snyder, D.B., W.W. Marquardt, E.T. Mallinson and E. Russek, 1983. Rapid serological profiling by Enzyme-linked immunosorbent assay. I. Measurement of antibody activity titer against Newcastle disease virus in a single serum dilution. Avian Dis., 27: 161-170.
    Direct Link    

  • OIE, 2004. Manual of diagnostic tests and vaccines for terrestrial animals (mammals, birds and bees). Office International des Epizooties, 5th Edn., Paris.

  • Varalakshmi, S., 2005. Assessment of cell mediated and mucosal immune response in chickens vaccinated with live Newcastle disease vaccines. M.V.Sc. Thesis, Tamil Nadu Vety. and Animal Sciences University Chennai.

  • Ward, M.D.W., F.J. Fuller, Y. Mehrotra and E.V. De Buysscher, 2000. Nucleotide sequence and vaccinia expression of the nucleoprotein of a highly virulent, neurotropic strain of Newcastle disease virus. Avian Dis., 44: 34-44.
    Direct Link    

  • Wilson, K. and J. Walker, 2006. Principles and Techniques of Biochemistry and Molecular Biology. 6th Edn., Cambridge University Press, Cambridge, UK., pp: 498-500

    • © Science Alert. All Rights Reserved