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J Virol Methods. 2009 Apr 27. [Epub ahead of print]
A Novel, Colorimetric Neutralization Assay for Measuring Antibodies to Influenza Viruses.
Lehtoranta L, Villberg A, Santanen R, Ziegler T. - National Influenza Center, Department of Vaccination and Immune Protection, National Institute for Health and Welfare, Mannerheimintie 166, 00300 Helsinki, Finland.
A colorimetric cell proliferation assay for measuring neutralizing antibodies to influenza viruses in human sera is described.
Following a 90-minute incubation, the serum-virus mixture was transferred to Madin-Darby canine kidney cells cultured in 96-well plates. After further incubation for three days, a tetrazolium salt was added to the wells. Cellular mitochondrial dehydrogenases cleave the tetrazolium salt to formazan, and the resulting color change is read by a spectrophotometer. The absorbance values correlate directly to the number of viable cells in the assay well and thus also to the neutralizing activity of influenza-specific antibodies present in the serum.
With the few hands-on manipulations required, this assay allows simultaneous testing of a considerable number of sera, offers opportunities for automation, and is suitable for use under biosafety level-3 conditions. The test was used to study the antibody response after administration of seasonal, inactivated, trivalent influenza vaccine.
Antibody titers determined by the neutralization test in pre- and post-vaccination serum pairs were compared with those obtained by the hemagglutination inhibition assay.
The neutralization test yielded higher pre- and post-vaccination titers and a larger number of significant increases in post-vaccination antibody titer than the hemagglutination inhibition test.
This new test format could serve as a valuable laboratory tool for influenza vaccine studies.
PMID: 19406159 [PubMed - as supplied by publisher]
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A Novel, Colorimetric Neutralization Assay for Measuring Antibodies to Influenza Viruses.
Lehtoranta L, Villberg A, Santanen R, Ziegler T. - National Influenza Center, Department of Vaccination and Immune Protection, National Institute for Health and Welfare, Mannerheimintie 166, 00300 Helsinki, Finland.
A colorimetric cell proliferation assay for measuring neutralizing antibodies to influenza viruses in human sera is described.
Following a 90-minute incubation, the serum-virus mixture was transferred to Madin-Darby canine kidney cells cultured in 96-well plates. After further incubation for three days, a tetrazolium salt was added to the wells. Cellular mitochondrial dehydrogenases cleave the tetrazolium salt to formazan, and the resulting color change is read by a spectrophotometer. The absorbance values correlate directly to the number of viable cells in the assay well and thus also to the neutralizing activity of influenza-specific antibodies present in the serum.
With the few hands-on manipulations required, this assay allows simultaneous testing of a considerable number of sera, offers opportunities for automation, and is suitable for use under biosafety level-3 conditions. The test was used to study the antibody response after administration of seasonal, inactivated, trivalent influenza vaccine.
Antibody titers determined by the neutralization test in pre- and post-vaccination serum pairs were compared with those obtained by the hemagglutination inhibition assay.
The neutralization test yielded higher pre- and post-vaccination titers and a larger number of significant increases in post-vaccination antibody titer than the hemagglutination inhibition test.
This new test format could serve as a valuable laboratory tool for influenza vaccine studies.
PMID: 19406159 [PubMed - as supplied by publisher]
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