Advertisement

Western immunoblotting as a method for the detection of African horse sickness virus protein-specific antibodies: Differentiation between infected and vaccinated horses

  • S. Idrissi Bougrine
  • O. Fassi Fihri
  • M. M. Fassi Fehri
Conference paper

Summary

A Western immunoblotting procedure has been developed for the detection of African horse sickness virus (AHSV) protein-specific antibody responses. This assay readily identifies antibodies specific for at least 4 distinct, AHSV proteins, including VP5, NS1, NS2 and NS3/NS3a. By using the AHSV non-structural proteins as ‘markers’, the Western blotting procedure could be employed to provide a reliable means of discriminating between animals vaccinated with a purified, inactivated AHSV vaccine and those either naturally infected or vaccinated with a live, attenuated AHSV vaccine.

Keywords

Nonstructural Protein Western Immunoblotting Antigen Preparation Bluetongue Virus Western Blotting Procedure 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Adkison MA, Stott JL, Osburn BI (1987) Identification of bluetongue virus protein-specific antibody responses in sheep by immunoblotting. Am J Vet Res 48: 1194–1198PubMedGoogle Scholar
  2. 2.
    Burrage TG, Travejo R, Stone-Marschat M, Laegreid WW (1993) Neutralising epitopes of African horse sickness virus serotype 4 are located on VP2. Virology 196: 799–803PubMedCrossRefGoogle Scholar
  3. 3.
    Burroughs JN, O’Hara RS, Smale CJ, Hamblin C, Walton A, Armstrong R, Mertens PPC (1994) Purification and properties of virus particles, infectious subviral particles, cores and VP7 crystals of African horse sickness virus serotype 9. J Gen Virol 75: 18491857Google Scholar
  4. 4.
    Dubourget P, Preaud JM, Detraz N, Lacoste F, Fabry AC, Erasmus BJ, Lombard M (1992) Development, production and quality control of an industrial inactivated vaccine against African horse sickness virus serotype 4. In: Walton TE, Osburn BI (eds) Bluetongue, African horse sickness and related orbiviruses. Proceedings of the Second International Symposium. Paris 1991. CRC Press, Boca Raton, pp 874–888Google Scholar
  5. 5.
    Grubman MJ, Lewis SA (1992) Identification and characterisation of the structural and non-structural proteins of African horse sickness virus and determination of the genome coding assignments. Virology 186: 444–451PubMedCrossRefGoogle Scholar
  6. 6.
    Herniman KAJ, Gumm ID, Owen L, Taylor WP, Sellers RF (1980) Distribution of bluetongue virus and antibodies in some countries of the eastern hemisphere. Bull Off Int Epiz 92: 581–586Google Scholar
  7. 7.
    Hess WR (1988) African horse sickness. In: Monath TP (ed) The arboviruses: epidemiology and ecology, vol 2. CRC Press, Boca Raton, pp 1–18Google Scholar
  8. 8.
    van Staden V, Smit CC, Stolz M, Maree FF, Huismans H (1998) Characterisation of two African horse sickness virus nonstructural proteins, NS3 and NS 1. In: Mellor PS, Baylis M, Hamblin C, Calisher CH, Mertens PPC (eds) African horse sickness. Springer, Wien New York, pp 251–258 (Arch Virol [Suppl] 14 )Google Scholar
  9. 9.
    House JA, Lombard M, Dubourget P, House C, Mebus CA (1994) Further studies on the efficacy of an inactivated African horse sickness serotype 4 vaccine. Vaccine 12: 142144Google Scholar
  10. 10.
    House JA (1998) Future international management of African horse sickness vaccines. In: Mellor PS, Baylis M, Hamblin C, Calisher CH, Mertens PPC (eds) African horse sickness. Springer, Wien New York, pp 297–304 (Arch Virol [Suppl] 14 )Google Scholar
  11. 11.
    Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685PubMedCrossRefGoogle Scholar
  12. 12.
    Laviada MD, Babin M, Dominguez J, Sanchez-Vizcaino JM (1992) Detection of African horse sickness in infected spleens by sandwich ELISA using two Monoclonal antibodies specific for VP7. J Virol Methods 38: 229–242PubMedCrossRefGoogle Scholar
  13. 13.
    Laviada MD, Arias M, Sanchez-Vizcaino JM (1993) Characterisation of African horse sickness virus serotype 4 induced polypeptides and their reactivity in Western immunoblotting. J Gen Virol 74: 81–87PubMedCrossRefGoogle Scholar
  14. 14.
    Laviada MD, Roy P, Sanchez-Vizcaino JM, Casal JI (1995) The use of African horse sickness virus NS3 protein, expressed in bacteria, as a marker to differentiate infected from vaccinated horses. Virus Res 38: 205–221PubMedCrossRefGoogle Scholar
  15. 15.
    Lhafi A, Tber A, Fikri A, Laghzaoui K (1992) African horse sickness in Morocco: the epizootics of 1989 and 1990. In: Walton TE, Osburn BI (eds) Bluetongue, African horse sickness and related orbiviruses. Proceedings of the Second International Symposium, Paris 1991. CRC Press, Boca Raton, pp 205–216Google Scholar
  16. 16.
    Lubroth J (1988) African horse sickness and the epizootic in Spain 1987. Equine Pract 10: 26–33Google Scholar
  17. 17.
    Mellor PS (1993) African horse sickness: transmission and epidemiology. Vet Res 24: 199–212PubMedGoogle Scholar
  18. 18.
    O’Hara RS (1994) Identification of the genome segments and proteins controlling the virulence of African horse sickness. PhD Thesis, Reading UniversityGoogle Scholar
  19. 19.
    O’Hara RS, Meyer AJ, Pullen L, Martin LA, Mertens PPC (1998) Development of a mouse model system and identification of the individual genome segments of African horse sickness virus serotypes 3 and 8 involved in determination of virulence. In: Mellor PS, Baylis M, Hamblin C, Calisher CH, Mertens PPC (eds) African horse sickness. Springer, Wien New York, pp 259–279 (Arch Virol [Suppl] 14 )Google Scholar
  20. 20.
    Ranz AI, Miguet JG, Anaya C, Venteo A, Cortes E, Vela C, Sanz A (1992) Diagnostic methods for African horse sickness virus using monoclonal antibodies to structural and non-structural proteins. Vet Microbiol 33: 143–153PubMedCrossRefGoogle Scholar
  21. 21.
    Roy P, Chuma T, Le Bois H, Hirasawa T (1992) Molecular cloning of RNA segment 7 and synthesis of the major core antigen VP7 of African horse sickness virus by recombinant Baculovirus: its use as a group-specific diagnostic reagent. In: Walton TE, Osburn BI (eds) Bluetongue, African horse sickness and related orbiviruses. Proceedings of the Second International Symposium, Paris 1991. CRC Press, Boca Raton, pp 461–470Google Scholar
  22. 22.
    Roy P (1998) New generation of African horse sickness vaccines based on structural and molecular studies of the virus particles. In: Mellor PS, Baylis M, Hamblin C, Calisher CH, Mertens PPC (eds) African horse sickness. Springer, Wien New York, pp 177–202 (Arch Virol [Suppl] 14 )Google Scholar
  23. 23.
    Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedures and some applications. Proc Natl Acad Sci USA 76: 4350–4354PubMedCrossRefGoogle Scholar
  24. 24.
    Van Staden V, Huismans H (1991) A comparison of the genes which encoded nonstructural protein NS3 of different Orbiviruses. J Gen Virol 72: 1073–1090PubMedCrossRefGoogle Scholar
  25. 25.
    Wade-Evans AM, Pullen L, Hamblin C, O’Hara RS, Burroughs JN, Mertens PPC (1998) VP7, from African horse sickness virus serotype 9, protects mice against a lethal heterologous serotype challenge. In: Mellor PS, Baylis M, Hamblin C, Calisher CH, Mertens PPC (eds) African horse sickness. Springer, Wien New York, pp 211–219 (Arch Virol [Suppl] 14 )Google Scholar

Copyright information

© Springer-Verlag/Wien 1998

Authors and Affiliations

  • S. Idrissi Bougrine
    • 1
  • O. Fassi Fihri
    • 2
  • M. M. Fassi Fehri
    • 2
  1. 1.Départment de Microbiologie Immunologie Maladies ContagieusesInstitut Agronomique et Vétérinaire Hassan IIRabatMorocco
  2. 2.Département de Microbiologie Immunologie Maladies ContagieusesInstitut Agronomique et Vétérinaire Hassan IIRabatMorocco

Personalised recommendations