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Neutralization assays for differential henipavirus serology using Bio-Plex protein array systems.

Katharine N Bossart1, Jennifer A McEachern, Andrew C Hickey

  • 1CSIRO Livestock Industries, Australian Animal Health Laboratory, 5 Portarlington Road, Geelong, Vic. 3220, Australia. Katharine.Bossart@csiro.au

Journal of Virological Methods
|February 13, 2007
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Summary

New assays can differentiate between Hendra virus (HeV) and Nipah virus (NiV) antibodies and detect neutralizing antibodies. These methods offer a safer alternative to live virus testing for henipaviruses.

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Area of Science:

  • Virology
  • Immunology
  • Biotechnology

Background:

  • Hendra virus (HeV) and Nipah virus (NiV) are emerging paramyxoviruses in the genus Henipavirus.
  • Both HeV and NiV cause severe, often fatal, diseases in humans and animals.
  • These viruses are classified as biosafety level 4 (BSL-4) pathogens, necessitating stringent safety measures for research.

Purpose of the Study:

  • To develop novel multiplexed microsphere assays for detecting and differentiating antibodies against HeV and NiV.
  • To create a surrogate assay for detecting virus neutralization without using live BSL-4 viruses.
  • To enable simultaneous detection and differentiation of HeV and NiV neutralizing antibodies.

Main Methods:

  • Development of two multiplexed microsphere assays utilizing recombinant soluble attachment glycoproteins (sG) of HeV and NiV.
  • One assay focused on antibody detection and differentiation; the second incorporated recombinant ephrin-B2 for neutralization surrogate testing.
  • Evaluation of antibody binding specificity and differential blocking of ephrin-B2 binding by henipavirus-seropositive sera and monoclonal antibodies (MAbs).

Main Results:

  • Spectrally distinct sG(HeV)- and sG(NiV)-coupled microspheres showed preferential binding to antibodies from HeV- and NiV-seropositive animals, enabling differentiation.
  • Recombinant ephrin-B2 binding to sG-coupled microspheres was dose-dependent and specificity was confirmed with henipavirus G-specific reagents.
  • Sera from seropositive animals differentially blocked ephrin-B2 binding, indicating potential for simultaneous detection of neutralizing antibodies.

Conclusions:

  • The developed multiplexed microsphere assays can effectively differentiate antibodies to the closely related HeV and NiV.
  • The ephrin-B2-based assay serves as a viable surrogate for detecting henipavirus-neutralizing antibodies.
  • These assays provide a safer, high-throughput method for diagnosing and monitoring henipavirus infections without live virus.