title: Neutralization assays for differential henipavirus serology using Bio-Plex protein array systems.
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.
authors.affiliation: CSIRO Livestock Industries, Australian Animal Health Laboratory, 5 Portarlington Road, Geelong, Vic. 3220, Australia. Katharine.Bossart@csiro.au
authors.forename: Katharine N
authors.initials: KN
authors.lastname: Bossart
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: McEachern
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Hickey
authors.forename: Vidita
authors.initials: V
authors.lastname: Choudhry
authors.forename: Dimiter S
authors.initials: DS
authors.lastname: Dimitrov
authors.forename: Bryan T
authors.initials: BT
authors.lastname: Eaton
authors.forename: Lin-Fa
authors.initials: LF
authors.lastname: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D002415:Cats; D036388:Ephrin-B2 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D045404:Hendra Virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D045403:Henipavirus / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D045464:Henipavirus Infections / Q000453:epidemiology / Q000821:virology; D006801:Humans; D051379:Mice; D008863:Microspheres; D009500:Neutralization Tests; D045405:Nipah Virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods; D011817:Rabbits; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012680:Sensitivity and Specificity; D012698:Serologic Tests; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036388:Ephrin-B2; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C116204:attachment protein G