Detection of bovine herpesvirus 1 with various types of DNA probes.
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.
Vilcek
S
S
Laboratory of Genetic Manipulations, Institute of Experimental Veterinary Medicine, Slovakia.
Deliová
I
I
Forgác
O
O
Strojný
L
L
Takácsová
I
I
Harvan
M
M
Benko
G
G
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D015342:DNA Probes / Q000737:chemistry; D004279:DNA, Viral / Q000737:chemistry / Q000302:isolation & purification; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D007242:Herpesvirus 1, Bovine / Q000235:genetics / Q000302:isolation & purification*; D015151:Immunoblotting; D008969:Molecular Sequence Data
D015342:DNA Probes; D004279:DNA, Viral