Initial experiences with consensus primer PCR for detection of animal herpesviruses (abstract)

Authors: Squires RA
Publication: New Zealand Veterinary Journal, Volume 53, Issue 1, pp 94, Feb 2005
Publisher: Taylor and Francis

Animal type: General
Subject Terms: DNA/RNA, Diagnostic procedures, Viral, Disease/defect, Infectious disease
Article class: Abstract

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Abstract: Consensus primer polymerase chain reaction (CP-PCR) is a useful technique for the detection and partial characterisation of novel viruses of veterinary importance. CP-PCR differs from conventional PCR, in that the upstream and downstream primers each consist of a mixture of slightly different oligonucleotides, rather than a single, defined sequence. As used in virus discovery, CP-PCR amplifies a segment of the genome of all members of a particular virus group. Conserved virus genes, e.g. those encoding enzymes, are the most suitable targets when designing these strategies. We used a previously validated CP-PCR method to amplify herpesviral DNA from the left conjunctival sac of a farmed red deer (Cervus elaphus) with keratitis. Sequencing of the CP-PCR product revealed 97% homology to a 174-base pair segment of a novel rhadinovirus of elk (Alces alces) (GenBank AY237365.1). An attempt to isolate a herpesvirus from the deer’s conjunctival sac in tissue culture was unsuccessful, as was PCR using conventional primers specific for Cervid herpesvirus 1. This case exemplifies the potential value of CP-PCR for detection of novel herpesviruses.

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