Bio-Protocol (Nov 2013)

Immunoplaque Assay (Influenza Virus)

  • Victor Tse,
  • Yueting Zhang,
  • Gary Whittaker

DOI
https://doi.org/10.21769/BioProtoc.959
Journal volume & issue
Vol. 3, no. 21

Abstract

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Despite developed long time ago, plaque assay is still the gold standard for viral titer quantification in modern virology. The standard crystal violet-based plaque assay relies on virus’ ability to induce cytopathic effect (CPE) which limits the assay to lytic viruses. Alternative viral quantification assays such as 50% tissue culture infectious assay (TCID50) and genetic material quantification by Q-PCR provide a different way of viral quantification with their own shortcoming. In here, we modified the fluorescent focus assay and developed an antibody-based immunoplaque assay which provides a reliable and reproducible viral quantification independent of CPE. Our assay not only allows accurate determination of viral titer, but also provides information on viral kinetics, genetic stability and purity of the virus population.