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J Immunol Res. 2014;2014:457932. doi: 10.1155/2014/457932. Epub 2014 Jul 3.

Multiplex evaluation of influenza neutralizing antibodies with potential applicability to in-field serological studies.

Author information

  • 1Viral Pseudotype Unit (Medway), School of Pharmacy, University of Kent, Central Avenue, Chatham Maritime, Kent ME4 4TB, UK.
  • 2Viral Pseudotype Unit (Fitzrovia), Faculty of Science and Technology, University of Westminster, London W1W 6UW, UK.
  • 3FAO, OIE, and National Reference Laboratory for Newcastle Disease and Avian Influenza, Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell'Università 10, 35020 Legnaro, Italy.

Abstract

The increased number of outbreaks of H5 and H7 LPAI and HPAI viruses in poultry has major public and animal health implications. The continuous rapid evolution of these subtypes and the emergence of new variants influence the ability to undertake effective surveillance. Retroviral pseudotypes bearing influenza haemagglutinin (HA) and neuraminidase (NA) envelope glycoproteins represent a flexible platform for sensitive, readily standardized influenza serological assays. We describe a multiplex assay for the study of neutralizing antibodies that are directed against both influenza H5 and H7 HA. This assay permits the measurement of neutralizing antibody responses against two antigenically distinct HAs in the same serum/plasma sample thus increasing the amount and quality of serological data that can be acquired from valuable sera. Sera obtained from chickens vaccinated with a monovalent H5N2 vaccine, chickens vaccinated with a bivalent H7N1/H5N9 vaccine, or turkeys naturally infected with an H7N3 virus were evaluated in this assay and the results correlated strongly with data obtained by HI assay. We show that pseudotypes are highly stable under basic cold-chain storage conditions and following multiple rounds of freeze-thaw. We propose that this robust assay may have practical utility for in-field serosurveillance and vaccine studies in resource-limited regions worldwide.

PMID:
25101305
PMCID:
PMC4101955
DOI:
10.1155/2014/457932
[PubMed - indexed for MEDLINE]
Free PMC Article
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