Bovine herpesvirus 1: rapid diagnosis of infection by direct filter hybridization.
Direct filter hybridization (DFH) was applied as a simple method of nucleic acid hybridization to diagnose bovine herpesvirus 1 (BHV-1) infection without previous purification of nucleic acids from the specimens. The DNA of BHV-1 was cleaved with the restriction endonuclease Pst I and randomly cloned into pKH47 plasmids. The clones were labelled with 32P or biotin and selected on uninfected and infected cells for the highest specific activity to detect BHV-1 infection. Two clones, which detected about 10 infected cells, were selected for the diagnosis of BHV-1 in cattle. On specimens collected during experimental and natural disease, the DFH showed to be in concordance with the standard method of virus isolation. This simple hybridization technique proved to be a sensitive and rapid alternative to virus isolation. Specific diagnosis of BHV-1 infection can be made even in simply equipped laboratories within 10 h.
Belák
S
S
Department of Virology, National Veterinary Institute, Uppsala, Sweden.
Linné
T
T
Magyar
G
G
Harrach
B
B
Benkö
M
M
Klingeborn
B
B
Klintevall
K
K
Bartha
A
A
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis; D007242:Herpesvirus 1, Bovine; D007241:Infectious Bovine Rhinotracheitis / Q000175:diagnosis*; D009693:Nucleic Acid Hybridization
D015342:DNA Probes; D004279:DNA, Viral