A rapid method for immunotitration of influenza viruses using flow cytometry

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 consumin...

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Bibliographic Details
Published in:Journal of virological methods 2003-06, Vol.110 (1), p.67-71
Main Authors: Lonsdale, R., Pau, M.G., Oerlemans, M., Ophorst, C., Vooys, A., Havenga, M., Goudsmit, J., UytdeHaag, F., Marzio, G.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cell Line
Dogs
Flow Cytometry - methods
Fundamental and applied biological sciences. Psychology
Humans
Influenza A virus - isolation & purification
Influenza A virus - metabolism
Influenza B virus - isolation & purification
Influenza B virus - metabolism
Influenza virus
MDCK
Method
Microbiology
Nucleocapsid Proteins
Nucleoproteins - metabolism
PER.C6
Plaque assay
Techniques used in virology
Time Factors
Titre
Viral Core Proteins - metabolism
Viral Plaque Assay
Virology
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Summary: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.
ISSN:0166-0934
1879-0984
DOI:10.1016/S0166-0934(03)00102-2