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Virus Genes

, Volume 18, Issue 3, pp 211–220 | Cite as

Detection of New DNA Polymerase Genes of Known and Potentially Novel Herpesviruses by PCR with Degenerate and Deoxyinosine-Substituted Primers

  • Bernhard Ehlers
  • Kerstin Borchers
  • Christian Grund
  • Kai Fro¨lich
  • Hanns Ludwig
  • Hans-Jo¨rg Buhk
Article

Abstract

A consensus primer PCR approach was used to (i) investigate the presence of herpesviruses in wild and zoo equids (zebra, wild ass, tapir) and to (ii) study the genetic relationship of the herpesvirus of pigeons (columbid herpesvirus 1) to other herpesvirus species. The PCR assay, based on degenerate primers targeting highly conserved regions of the DNA polymerase gene of herpesviruses, was modified by using a mixture of degenerate and deoxyinosine-substituted primers. The applicability of the modification was validated by amplification of published DNA polymerase genes of 16 herpesvirus species and of the previously uncharacterized DNA polymerase genes of equine herpesvirus 3 (EHV-3) and equine herpesvirus 5 (EHV-5). The modified assay was then used for partial amplification of the polymerase of columbid herpesvirus 1 which is presently classified as a β-herpesvirus based on biological criteria. Sequence analysis of amplicons obtained from four different viral strains revealed a close relationship of columbid herpesvirus 1 to members of the subfamily Alphaherpesvirinae, especially to Marek’s disease herpesvirus. This was confirmed by characterization of additional 1.6 kb of the columbid herpesvirus 1 polymerase. Consensus PCR analysis of blood samples from zebras, a wild ass and a tapir revealed amplicons showing high percentages (>50%) of sequence identity to DNA polymerases of γ-herpesviruses. In particular, the zebra and the wild ass sequence were closely related to each other and to the polymerases of the equine γ-herpesviruses EHV-2 and EHV-5 with sequence identities of >80%. This is a first indication that novel γ-herpesviruses are present in wild and zoo equids.

herpesviridae DNA polymerase consensus primer deoxyinosine equine herpesviruses columbid herpesvirus 1 

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Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Bernhard Ehlers
    • 1
  • Kerstin Borchers
    • 2
  • Christian Grund
    • 3
  • Kai Fro¨lich
    • 4
  • Hanns Ludwig
    • 4
  • Hans-Jo¨rg Buhk
    • 1
  1. 1.Robert Koch-InstitutBerlin
  2. 2.Institut fu¨r Virologie, Fachbereich Veterina¨rmedizinFreie Universita¨t BerlinBerlin
  3. 3.Institut fu¨r Geflu¨gelkrankheitenLudwig-Maximilians-Universita¨tOberschleißheim
  4. 4.Institut fu¨r Zoo und WildtierforschungBerlin

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