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Detection of bovine herpesvirus 1 with various types of DNA probes
S Vilcek1, I Deliová, O Forgác
1Laboratory of Genetic Manipulations, Institute of Experimental Veterinary Medicine, Slovakia.
Abstract:
The method of dot-blot hybridization on nitrocellulose filters by various types of DNA probes (ds recombinant plasmids, ss recombinant M13 phages and a 42bp synthetic oligonucleotide) was used for BHV-1 detection. The highest sensitivity was achieved with the 32P-pUR1 probe (1.8 kb random EcoRI-HindIII fragment inserted into pUC9) which detected the BHV-1 genome in 5 x 10(3) infected MDBK cells. Using the pUR1 probe, no cross hybridization was observed with other herpesviruses: BHV-2, 3, 4, and Aujeszky's disease virus. The 32P-pUR1 probe detected BHV-1 in nasal swabs of calves as early as on day 1 after experimental infection. The maximum intensity of BHV-1 detection occurred on day 1-3. The 32P-pUR1 probe also detected BHV-1 in field samples of nasal swabs from cows and calves.