Neutralization assays for differential henipavirus serology using Bio-Plex protein array systems.
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.
Bossart
Katharine N
KN
CSIRO Livestock Industries, Australian Animal Health Laboratory, 5 Portarlington Road, Geelong, Vic. 3220, Australia. Katharine.Bossart@csiro.au
McEachern
Jennifer A
JA
Hickey
Andrew C
AC
Choudhry
Vidita
V
Dimitrov
Dimiter S
DS
Eaton
Bryan T
BT
Wang
Lin-Fa
LF
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
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036388:Ephrin-B2; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C116204:attachment protein G