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2007 | 63 | 06 |

Tytuł artykułu

Zastosowanie testu immunoperoksydazowego posredniego do potwierdzania tozsamosci izolatow EAV

Autorzy

Warianty tytułu

EN
Application of the indirect immunoperoxidase test to confirm the specificity EAV isolates

Języki publikacji

PL

Abstrakty

EN
The purpose of the study was the elaboration and introduction of an effective method to confirm the identity of EAV isolates obtained in cell culture from the serum of persistently infected stallions. In view of the low cost, simplicity of preparation, and its high specificity, an indirect immunoperoxidase test based on rabbit polyclonal sera was introduced. After immunization of the rabbits with the EAV reference strain Bucyrus, purified by ultracentrifugation, a specific polyclonal serum in the titer of SN antibodies from 1:32 to 1:128 was obtained. Peroxidase-conjugated goat IgG antibodies against horse immunoglobulins were used as a secondary antibody and DAB as a substrate. After the optimalization of the test conditions, its sensitivity and specificity was defined. The sensitivity estimated in reference to the Bucyrus strain was equal to 1 TCID₅₀. The specificity was defined at 100%. After the preliminary validation, the indirect immunoperoxidase test based on the obtained polyclonal sera was affirmed as a method to identify EAV isolates. All 68 EAV field isolates reacted positively in the immunoperoxidase test.

Wydawca

-

Rocznik

Tom

63

Numer

06

Opis fizyczny

s.712-716,rys.,fot.,tab.,bibliogr.

Twórcy

autor
  • Panstwowy Instytut Weterynaryjny, Al.Partyzantow 57, 24-100 Pulawy
autor

Bibliografia

  • 1. Balasuriya U. B. R., Evermann J. F., Hedges J. F., McKeirnan A. J., Mitten J. Q., Beyer J. C., McCollum W. H., Timoney P. J., MacLachlan N. J.: Serologic and molecular characterization of an abortigenic strain of equine arteritis virus isolated from infective frozen semen and an aborted equine fetus. J. Am. Vet. Med. Assoc. 1998, 213, 1586-1589.
  • 2. Balasuriya U. B. R., Snijder E. J., van Dinten L. C., Heidner H. W., Wilson W. D., Hedges J. F., Hullinger P. J., MacLachlan N. J.: Equine arteritis virus derived from an infectious cDNA clone is attenuated and genetically stable in infected stallions. Virology 1999, 260, 201-208.
  • 3. Balasuriya U. B. R., Timoney P. J., McCollum W. H., MacLachlan N. J.: Phylogenetic analysis of open reading frame 5 of field isolates of equine arteritis virus and identification of conserved and nonconserved regions in the GL envelope glycoprotein. Virology 1995, 214, 690-697.
  • 4. Deregt D., de Vries A. A., Raamsman M. J., Elmgren L. D., Rottier P. J.: Monoclonal antibodies to equine arteritis virus proteins identify the GL protein as a target for virus neutralization. J. Gen. Virol. 1994, 75, 2439-2444.
  • 5. Doll E. R., Knappenberger R. E., Bryans J. T.: An outbreak of abortion caused by the equine arteritis virus. Cornell Vet. 1957, 47, 69-75.
  • 6. Golnik W., Michalak T.: Przypadki wirusowego zapalenia têtnic koni (arteritis equorum) w Polsce. Medycyna Wet. 1978, 35, 605-606.
  • 7. Golnik W., Moraillon A., Golnik J.: Identification and antigenic comparison of equine arteritis virus isolated from an outbreak of epidemic abortion of mares. J. Vet. Med. Ser. B 1986, 33, 413-417.
  • 8. Hornyak A., Denes B., Szeredi L., Dencso L., Rusvai M.: Diagnostic application of immunoperoxidase monolayer assay using monoclonal antibodies produced against equine arteritis virus 14-kDa nucleocapsid protein. Hybrid Hybridomics 2004, 23, 368-372.
  • 9. Little T. V., Holyoak G. R., McCollum W. H., Timoney P. J.: Output of equine arteritis virus from persistently infected stallions is testosterone-dependent. 1992, 225, Plowright W., Rossdale P. D., Wada J. F. (red.), Proceedings of the 6th International Conference on Equine Infectious Diseases, Cambridge 1991, R&W Publications Ltd., Newmarket.
  • 10. Murphy T. W., McCollum W. H., Timoney P. J., Klingeborn B. W., Hyllseth B., Golnik W., Erasmus B.: Genomic variability among globally distributed isolates of equine arteritis virus. Vet. Microbiol. 1992, 32, 101-115.
  • 11. Raś A., Siedlanowski T.: Badać czy nie badać? Koń Polski 2001, 2, 50-51.
  • 12. Starick E.: Rapid and sensitive detection of equine arteritis virus in semen and tissue samples by reverse transcription-polymerase chain reaction, dot blot hybridisation and nested polymerase chain reaction. Acta Virol. 1998, 42, 333-339.
  • 13. Timoney P. J.: OIE Manual of diagnostic tests and vaccines for terrestrial animals: Equine arteritis virus. 2004, rozdz. 2.5.10, 725-736.
  • 14. Timoney P. J., McCollum W. H., Murphy T. W., Roberts A. W., Willard A. W., Carswell J. G.: The carrier state in equine arteritis virus infection in the stallion with specific emphasis on the venereal mode of transmission. J. Reprod. Fertil. Suppl. 1987, 35, 95-102.
  • 15. Timoney P. J., McCollum W. H., Roberts A. W., Murphy T. W.: Demonstration of the carrier state in natural acquired equine arteritis virus infection in the stallion. Res. Vet. Sci. 1986, 41, 279-280.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

bwmeta1.element.agro-article-53b86280-4b7e-4ad8-9468-c205109baefe
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