Archives of Virology

, Volume 71, Issue 2, pp 177–183 | Cite as

A modified plaque assay method for accurate analysis of infectivity of influenza viruses with uncleaved hemagglutinin

  • O. P. Zhirnov
  • A. V. Ovcharenko
  • A. G. Bukrinskaya
Original Papers


A modified plaque assay method for evaluation of infectivity of influenza viruses is described. The modification consists of a double agar overlay, trypsin being included in the second layer added 24 hours after infection. The modified method allows one to determine accuratelly the infectivity titer of influenza viruses possessing uncleaved hemagglutinin, and to compare the infectivity of viruses possessing cleaved or uncleaved forms of hemagglutinin. The standard method in which trypsin is included into agar overlay just after infection artifically increases the infectivity titer of viruses with uncleaved hemagglutinin and therefore is not suitable for these purposes.


Agar Infectious Disease Influenza Trypsin Standard Method 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Appleyard, G., Maber, H. B.: Plaque formation by Influenza Viruses in the presence of trypsin. J. gen. Virol.25, 351–357 (1974).Google Scholar
  2. 2.
    Came, P. E., Pascale, E., Shimonaski, G.: Effect of pancreatin on plaque formation by Influenza Viruses. Arch. ges. Virusforsch.23, 346–352 (1968).Google Scholar
  3. 3.
    Homma, M., Ohuchi, M.: Trypsin action on the growth on Sendai virus in tissue culture cells. III. Structural difference of Sendai viruses grown in eggs and tissue culture cells. J. Virol.12, 1457–1465 (1973).Google Scholar
  4. 4.
    Huang, R. T. C., Wahn, K., Klenk, H.-D., Rott, R.: Fusion between cell membrane and liposomes containing the glycoproteins of influenza virus. Virology104, 294–302 (1980).Google Scholar
  5. 5.
    Huang, R. T. C., Rott, R., Wahn, K., Klenk, H.-D., Kohama, T.: The function of the neuraminidase in membrane fusion induced by myxoviruses. Virology107, 313–319 (1980).Google Scholar
  6. 6.
    Klenk, H.-D., Rott, R., Orlich, M., Blödorn, J.: Activation of influenza viruses by trypsin treatment. Virology68, 426–439 (1975).Google Scholar
  7. 7.
    Kurrle, R., Wagner, H., Röllinghoff, M., Rott, R.: Influenza virus-specific T cell-mediated cytotoxicity: Integration of the virus antigen into the target cell membrane is essential for target cell formation. Eur. J. Immunol.9, 107–111 (1979).Google Scholar
  8. 8.
    Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature (London)227, 680–685 (1970).Google Scholar
  9. 9.
    Laver, W. G.: Separation of two polypeptides chains from the hemagglutinin subunit of influenza virus. Virology45, 275–278 (1971).Google Scholar
  10. 10.
    Lazarowitz, S. G., Goldberg, A. R., Choppin, P. W.: Proteolytic cleavage by plasmin of the HA polypeptide of influenza virus. Host cell activation of serum plasminogen. Virology56, 172–180 (1973).Google Scholar
  11. 11.
    Lazarowitz, S. G., Choppin, P. W.: Enhancement of the infectivity of influenza A and B viruses by proteolytic cleavage of the hemagglutinin polypeptides. Virology68, 440–454 (1975).Google Scholar
  12. 12.
    Nagai, Y., Klenk, H.-D., Rott, R.: Proteolytic cleavage of the viral glycoproteins and its significance for the virulence of Newcastle disease virus. Virology72, 494–508 (1976).Google Scholar
  13. 13.
    Nicolau, C., Klenk, H.-D., Reiman, A., Hildebrand, K., Bauer, H.: Molecular events during the interaction of envelopes of myxo- and RNA tumor viruses with cell membranes. A 270 MHz1H nuclear magnetic resonance study. Biochem. Biophys. Acta511, 83–92 (1979).Google Scholar
  14. 14.
    Scheid, A., Choppin, P. W.: Identification of biological activities of paramyxovirus glycoproteins. Activation of cell fusion, hemolysis and infectivity by proteolytic cleavage of an inactive precursor protein of Sendai virus. Virology57, 475 to 490 (1974).Google Scholar
  15. 15.
    Skehel, J. J., Waterfield, M. D.: Studies on primary structure of the influenza virus hemagglutinin. Proc. Nat. Acad. Sci. U.S.A.72, 93–97 (1975).Google Scholar
  16. 16.
    Tobita, K., Kilbourne, E. D.: Genetic recombination for antigenic markers of antigenically different strains of influenza B virus. J. Virol.13, 347–352 (1974).Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • O. P. Zhirnov
    • 1
  • A. V. Ovcharenko
    • 1
  • A. G. Bukrinskaya
    • 1
  1. 1.The D. I. Ivanovsky Institute of VirologyThe U.S.S.R. Academy of Medical SciencesMoscowU.S.S.R.

Personalised recommendations