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


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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  1. 1.
    Borchers K. and FroÈlich K., J Wildlife Dis 33, 812-817, 1997Google Scholar
  2. 2.
    Messana M., Kuesters J., and Grund C., Avian Pathology 26, 859-864, 1997Google Scholar
  3. 3.
    Ziemann K., Mettenleiter T.C., and Fuchs W., J Virol 72, 847-852, 1998Google Scholar
  4. 4.
    Buckmaster A.E., Scott S.D., Sanderson M.J., Boursnell M.E.G., Ross N.L.J., and Binns M.M., J Gen Virol 69, 2033-2042, 1988Google Scholar
  5. 5.
    Kaleta E.F., Avian Pathology 26, 305-316, 1997Google Scholar
  6. 6.
    VanDevanter D.R., Warrener P., Bennett L., Schultz E.R., Coulter S., Garber R.L., and Rose T.M., J Clin Microbiol 34, 1666-1671, 1996Google Scholar
  7. 7.
    Ehlers B., Buhk H.-J., and Ludwig H., J Gen Virol 66, 55-68, 1985Google Scholar
  8. 8.
    Kebelmann-Betzing C., Seeger K., Dragon S., Schmitt G., MoÈricke A., Schild T.A., Henze G., and Beyermann B., BioTechniques 24, 154-158, 1998Google Scholar
  9. 9.
    Rossolini G.M., Cresti S., Ingianni A., Cattani P., Riccio M.L., and Satta S., Mol Cell Probes 8, 91-98, 1994Google Scholar
  10. 10.
    Knoth K., Roberds S., Poteet C., and Tamkun M., Nucl Acids Res 16, 10932, 1988Google Scholar
  11. 11.
    Cassol S., Salas T., Lapointe N., Arella M., Rudnik J., and O'Shaughnessy M., Mol Cell Probes 5, 157-160, 1991Google Scholar
  12. 12.
    McGeoch D.J., Cook S., Dolan A., Jamieson F.E., and Telford E.A.R., J Mol Biol 247, 443-458, 1995Google Scholar
  13. 13.
    Rovnak J., Quackenbush S.L., Reyes R.A., Baines J.D., Parrish, C.R., and Casey J.W., J Virol, 72, 4237-4242, 1998Google Scholar
  14. 14.
    Cornwell H.J.C. and Wright N.G., J Comp Path 80, 221-226, 1970Google Scholar
  15. 15.
    Sallmann M., (Diss. vet. med.), University of Giessen, 1991Google Scholar
  16. 16.
    Knopf C.W., Virus Genes 16, 47-58, 1998Google Scholar
  17. 17.
    Desrosiers R.C., Sasseville V.G., Czajak S.C., Zhang X., Mans®eld K.G., Kaur A., Johnson P., Lackner A.A., and Jung J.U., J Virol 71, 9764-9769, 1998Google Scholar
  18. 18.
    Borchers K., Wol®nger U., Goltz M., Broll H., and Ludwig H., Arch Virol, 142, 917-928, 1997Google Scholar
  19. 19.
    Browning C.F. and Studdert M.J., Vet Bull 58, 775-790, 1988Google Scholar
  20. 20.
    Telford E.A.R., Studdert M.J., Agius C.T., Watson M.S., Aird H.C., and Davison A.J., Virology 195, 492-499, 1993Google Scholar

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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