Skip to main content
Log in

A comparison of whole virus and recombinant transmembrane ELISA and immunodiffusion for detection of ovine lentivirus antibodies in Italian sheep flocks

  • Virology
  • Review Article
  • Published:
Veterinary Research Communications Aims and scope Submit manuscript

Abstract

Sera from two sheep experimentally infected with ovine lentivirus (OLV) and from 186 sheep selected from flocks with known high or low prevalence of infection or on the basis of virological or histopathological examination were simultaneously tested by whole virus (WV) ELISA, recombinant transmembrane (r-TM) ELISA and AGID assay. Antigens for both the WV ELISA and AGID were prepared from an Italian field isolate; recombinant antigen was derived from the N′-terminal region of the transmembrane envelope protein of strain K1514. The WV ELISA detected the highest number of seropositives, followed by the r-TM ELISA and AGID test. The sensitivity and specificity of the r-TM ELISA relative to the WV ELISA were 0.66 and 0.95, respectively. Immunoblot analysis of 14 WV ELISA-positive and r-TM ELISA-negative sera showed that the major core protein was immunodominant on WV antigen. It is concluded that the r-TM ELISA was more sensitive than the AGID test but less sensitive that the WV ELISA, particularly for detecting antibodies in the early stages of infection.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

AGID:

agar gel immunodiffusion

bp:

base pair

ELISA:

enzyme-linked immunosorbent assay

FBS:

fetal bovine serum

IB:

immunoblot

kD:

kilodalton

OLV:

ovine lentivirus

MEM:

minimal essential medium

p.i.:

post-infection

r:

recombinant

SDS-PAGE:

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

TM:

transmembrane

WV:

whole virus

References

  • Burnet, W.N., 1981. “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.Analytical Biochemistry,112, 195–203

    Google Scholar 

  • Cutlip, R.C., Jackson, T.A. and Lehmkuhl, H.D., 1977. Immunodiffusion test for ovine progressive pneumonia.American Journal of Veterinary Research,38, 1081–1084

    Google Scholar 

  • De Savigny, D. and Voller, A., 1980. The communication of ELISA data from laboratory to clinician.Journal of Immunoassay,1, 105–128

    Google Scholar 

  • Fleiss, J.L., 1981.Statistical Methods for Rates and Proportions, 2nd edn, (J Wiley, New York), 212–236

    Google Scholar 

  • Houwers, D.J. and Nauta, I.M., 1989. Immunoblot analysis of the antibody response to ovine lentivirus infections.Veterinary Microbiology,19, 127–139

    Google Scholar 

  • Houwers, D.J., Gielkens, S.L.J. and Schaake, J., 1982. An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to maedi-visna virus.Veterinary Microbiology,7, 209–219

    Google Scholar 

  • Kwang, J. and Cutlip, R.C., 1992a. Analysis of antibody response to ovine lentivirus by using viral gene products expressed in a prokaryotic system.Biochemical and Biophysical Research Communications,188, 20–27

    Google Scholar 

  • Kwang, J. and Cutlip, R.C., 1992b. Detection of antibodies to ovine lentivirus using a recombinant antigen derived from theenv gene.Biochemical and Biophysical Research Communications,183, 1040–1046

    Google Scholar 

  • Kwang, J., Keen, J., Cutlip, R.C. and Littledike, E.T., 1993. Evaluation of an ELISA for detection of ovine progressive pneumonia antibodies using a recombinant transmembrane envelope protein.Journal of Veterinary Diagnostic Investigation,5, 189–193

    Google Scholar 

  • Kwang, J., Keen, J., Cutlip, R.C., Kim, H.S. and Concha-Bermejillo, A., 1994. Serological diagnosis of caprine lentivirus infection by recombinant immunoassays.Journal of Small Ruminant Research, In press

  • Laemmli, U., 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature,227, 680–685

    Google Scholar 

  • Querat, G., Audoly, G., Sonigo, P., Vigne, R., 1990. Nucleotide sequence analysis of SA-OMVV, a visna related ovine lentivirus: phylogenetic history of lentiviruses.Virology,175, 434–447

    Google Scholar 

  • Rosati, S., Tolari, F. and Bollo, E., 1992. Identification of lentiviruses isolated from outbreaks of ovine maedi and caprine arthritis encephalitis.Proceedings, Annual Meeting of the Italian Society of Veterinary Science,46, 883–886

    Google Scholar 

  • Sargan, D.R., Bennet, I.D., Cousens, C., Roy, D.J., Blacklaws, B.A., Dalziel, R.G., Watt, N.J. and McConnell, I., 1991. Nucleotide sequence of EV1, a British isolate of maedi-visna virus.Journal of General Virology,72, 1893–1903

    Google Scholar 

  • Sonigo, P., Alizon, M., Staskus, K., Klatzmann, D., Cole, S., Danos, O., Retzel, E., Tiollais, P., Haase, A. and Wain-Hobson, S., 1985. Nucleotide sequence of the visna lentivirus: relationship to the AIDS virus.Cell,42, 369–382

    Google Scholar 

  • Tolari, F., Rosati, S., Valenza, F., Braca, G. and Legrottaglie, R., 1992. Ovine maedi: correlation among serological, virological and anatomo-histopathological investigations in sheep from flocks with low prevalence of infection.Proceedings, Annual Meeting of the Italian Society of Veterinary Science,46, 887–891

    Google Scholar 

  • Torfason, G.E., Gudnadottir, M. and Love, A., 1991. Comparison of immunoblots with neutralizing and complement fixing antibodies in experimental and natural cases of visna-maedi.Archives of Virology,123, 47–58

    Google Scholar 

  • Vitu, C., Russo, P., Filippi, P., Vigne, R., Querat, G. and Giauffret, A., 1982. Une Technique Elisa pour la Detection des Anticorps anti-virus Maedi-Visna. Etude comparative avec l'Immunodiffusion en Gelose et la Fixation du Complement.Comparative Immunology, Microbiology and Infectious Diseases,5, 469–481

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rosati, S., Kwang, J., Tolari, F. et al. A comparison of whole virus and recombinant transmembrane ELISA and immunodiffusion for detection of ovine lentivirus antibodies in Italian sheep flocks. Vet Res Commun 18, 73–80 (1994). https://doi.org/10.1007/BF01839262

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01839262

Keywords

Navigation