Abstract
Hendra virus (HeV) and Nipah virus (NiV) are related emerging paramyxoviruses classified in the genus Henipavirus. Both cause fatal disease in animals and humans and are classified as biosafety level 4 pathogens. Here we detail two new multiplexed microsphere assays, one for antibody detection and differentiation and another designed as a surrogate for virus neutralization. Both assays utilize recombinant soluble attachment glycoproteins (sG) whereas the latter incorporates the cellular receptor, recombinant ephrin-B2. Spectrally distinct sG(HeV)- and sG(NiV)-coupled microspheres preferentially bound antibodies from HeV- and NiV-seropositive animals, demonstrating a simple procedure to differentiate antibodies to these closely related viruses. Soluble ephrin-B2 bound sG-coupled microspheres in a dose-dependent fashion. Specificity of binding was further evaluated with henipavirus G-specific sera and MAbs. Sera from henipavirus-seropositive animals differentially blocked ephrin-B2 binding, suggesting that detection and differentiation of HeV and NiV neutralizing antibodies can be done simultaneously in the absence of live virus.
Publication types
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Evaluation Study
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Research Support, N.I.H., Intramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal / immunology
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Antibodies, Viral / blood*
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Antibodies, Viral / immunology
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Cats
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Ephrin-B2 / genetics
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Ephrin-B2 / immunology
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Ephrin-B2 / metabolism
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Hendra Virus / genetics
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Hendra Virus / immunology*
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Hendra Virus / metabolism
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Henipavirus / genetics
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Henipavirus / immunology
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Henipavirus / metabolism
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Henipavirus Infections / epidemiology
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Henipavirus Infections / virology
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Humans
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Mice
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Microspheres
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Neutralization Tests*
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Nipah Virus / genetics
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Nipah Virus / immunology*
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Nipah Virus / metabolism
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Protein Array Analysis* / instrumentation
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Protein Array Analysis* / methods
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Rabbits
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Reagent Kits, Diagnostic*
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Recombinant Proteins / genetics
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Recombinant Proteins / immunology
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Recombinant Proteins / metabolism
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Sensitivity and Specificity
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Serologic Tests
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Viral Envelope Proteins / genetics
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Viral Envelope Proteins / immunology
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Viral Envelope Proteins / metabolism
Substances
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Antibodies, Monoclonal
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Antibodies, Viral
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Ephrin-B2
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Reagent Kits, Diagnostic
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Recombinant Proteins
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Viral Envelope Proteins
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attachment protein G