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

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

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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