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  • Title: A rapid method for immunotitration of influenza viruses using flow cytometry.
    Author: Lonsdale R, Pau MG, Oerlemans M, Ophorst C, Vooys A, Havenga M, Goudsmit J, UytdeHaag F, Marzio G.
    Journal: J Virol Methods; 2003 Jun 09; 110(1):67-71. PubMed ID: 12757922.
    Abstract:
    Reliable assays for accurate titration of influenza virus in infectious samples are pivotal to both influenza research and vaccine development. A titration assay adopted commonly for this purpose is the plaque assay on Madin-Darby canine kidney (MDCK) cells, despite it being time and labour consuming. A novel assay is described for titration of influenza viruses based on the detection of intracellular viral nucleoprotein (NP) by fluorescence-activated cell sorting (FACS). By using a panel of viruses of different type, subtype and origin, it is demonstrated that there is a mathematical correlation between titres measured by immunotitration and by classical plaque assay on MDCK cells. Moreover, the availability of NP antibodies specific for type A or type B influenza virus ensures the specificity of the assay. Based on speed, accuracy and specificity, it is concluded that the FACS-based immunotitration of influenza virus represents a valid and efficient alternative to the classical plaque assay.
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