Abstract
It may take several months before goats and sheep infected with lentiviruses develop measurable antiviral antibodies. During this period, virus may be isolated from infected animals but current cell culture techniques for the isolation and identification of goat and sheep lentiviruses are tedious and take several weeks to complete. Using the polymerase chain reaction, we show here that virus can be detected in infected cell cultures with high specificity and sensitivity. To demonstrate both CAE- and Maedi-Visna virus-specific DNA we utilized primers 20 base pairs long and framing 215 nucleotides of the gag gene (p28/p11 region) of Visna virus. DNA of Maedi-Visna and CAE viruses could be detected at 1 day post-infection in cultured sheep plexus and lamb carpal cells, respectively. This was 4 days before CAE viral proteins could be demonstrated by immunoblotting. The PCR was highly sensitive, allowing the detection of approximately one infected cell in a culture containing 10 6 cells. Our experiments indicate that the PCR exceeds the sensitivity of conventional detection methods for Maedi-Visna and CAE viruses. Experiments to adapt the PCR to material obtained from infectes animals are under way.
Published Version
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