Detection of bovine immunodeficiency-like virus infection in experimentally infected calves |
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| Authors: | T Baron D Betemps F Mallet V Cheynet D Levy P Belli |
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| Institution: | (1) Centre National d’Etudes Vétéerinaires et Alimentaires, Lyon, France, FR;(2) UMR 103 CNRS-BioMérieux, Ecole Normale Supéerieure, Lyon, France, FR;(3) URA INRA “Immunopathologie cellulaire et moléeculaire”, Maisons-Alfort, France, FR |
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| Abstract: | Summary. Detection of BIV virus infection by serological means, PCR and virus isolation in experimentally infected calves is described.
Viral sequences were specifically detected by PCR in peripheral blood mononuclear cells (PBMCs), with primer systems located
in the gag, pol and tat regions of the viral genome. An enzyme-linked oligosorbent assay (ELOSA) in microtiter plates is described, for the detection
of PCR products, the sensitivity of which was shown to be comparable to that of membrane hybridization detection. Serological
response of the animals against the BIV p26 protein was shown, using a recombinant fusion protein ((His)6p26) expressed in E. coli and purified by metal affinity chromatography, in ELISA and Western blot studies. The presence of infectious virus was demonstrated
by its rescue, by virus isolation in
cell cultures, from PBMCs during a one year follow-up.
Accepted August 14, 1997 Received November 28, 1996 |
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