Detection of antibodies to ovine lentivirus using recombinant capsid and transmembrane proteins.
The coding sequences of the capsid protein p25 and transmembrane protein of Maedi-Visna virus were amplified using polymerase chain reaction and cloned into the plasmid expression vector pRSET-B. Both DNA constructs expressed proteins tagged with polyhistidine. The recombinant proteins were purified using Ni-NTA agarose and used in immunoblot to detect antibodies against Maedi-Visna virus. A total of 260 ovine serum specimens was analysed. The total number of p25-positive sera was 111 (42.7%). Higher sensitivity was achieved with rTM antigen, which detected antibodies in 118 (45.4%) sera. The combination of both recombinant proteins as antigens resulted in higher sensitivity of serological detection compared to whole virus antigen.
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Department of Microbiology and Immunology, University of Veterinary and Pharmaceutical Sciences Brno, Palackého 1/3, CZ-612 42 Brno, Czech Republic.
Celer
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D000818:Animals; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology; D015153:Blotting, Western; D002213:Capsid / Q000276:immunology*; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D008565:Membrane Proteins / Q000276:immunology; D011021:Pneumonia, Progressive Interstitial, of Sheep / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011237:Predictive Value of Tests; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D012756:Sheep; D014763:Viral Matrix Proteins / Q000276:immunology*; D014790:Visna-maedi virus / Q000276:immunology* / Q000302:isolation & purification
D000914:Antibodies, Viral; D000956:Antigens, Viral; D017931:DNA Primers; D008565:Membrane Proteins; D011994:Recombinant Proteins; D014763:Viral Matrix Proteins