Bovine herpesvirus 1: rapid diagnosis of infection by direct filter hybridization

S. Belák, T. Linné, G. Magyar, B. Harrach, Mária Benkö, B. Klingeborn, Kristina Klintevall, A. Bartha

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Abstract

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.

Original languageEnglish
Pages (from-to)147-156
Number of pages10
JournalMolecular and Cellular Probes
Volume2
Issue number2
DOIs
Publication statusPublished - Jun 1988

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Keywords

  • bovine herpesvirus 1
  • direct filter hybridization
  • infectious bovine rhinotracheitis
  • nucleic acid hybridization

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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