title: Novel multiplex assay platforms to detect influenza A hemagglutinin subtype-specific antibody responses for high-throughput and in-field applications.
abstract: Detections of influenza A subtype-specific antibody responses are often complicated by the presence of cross-reactive antibodies. We developed two novel multiplex platforms for antibody detection. The multiplexed magnetic fluorescence microsphere immunoassay (MAGPIX) is a high-throughput laboratory-based assay. Chembio Dual Path Platform (DPP) is a portable and rapid test that could be used in the field.
  
  
  
  Twelve recombinant globular head domain hemagglutinin (GH HA1) antigens from A(H1N1)pdm09 (pH1N1), A(H2N2), A(H3N2), A(H5N1), A(H7N9), A(H9N2), A(H13N9), B/Victoria lineage, B/Yamagata lineage viruses, and protein A control were used. Human sera from U.S. residents either vaccinated (with H5N1 or pH1N1) or infected with pH1N1 influenza viruses and sera from live bird market workers in Bangladesh (BDPW) were evaluated. GH HA1 antigens and serum adsorption using full ectodomain recombinant hemagglutinins from A(pH1N1) and A(H3N2) were introduced into the platforms to reduce cross-reactivity.
  
  
  
  Serum adsorption reduced cross-reactivity to novel subtype HAs. Compared to traditional hemagglutination inhibition or microneutralization assays, when serum adsorption and the highest fold rise in signals were used to determine positivity, the correct subtype-specific responses were identified in 86%-100% of U.S. residents exposed to influenza antigens through vaccination or infection (N=49). For detection of H5N1-specific antibodies in sera collected from BDPW, H5 sensitivity was 100% (six of six) for MAGPIX, 83% (five of six) for DPP, H5 specificity was 100% (15/15), and cross-reactivity against other subtype was 0% (zero of six) for both platforms.
  
  
  
  MAGPIX and DPP platforms can be utilized for high-throughput and in-field detection of novel influenza virus infections.
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Zhu-Nan
authors.identifier: 0000-0002-4556-8551
authors.initials: ZN
authors.lastname: Li
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.|Department of Microbiology and Immunology, Emory University, Atlanta, GA, USA.
authors.forename: Jessica F
authors.initials: JF
authors.lastname: Trost
authors.affiliation: Battelle Memorial Institute, Columbus, OH, USA.
authors.forename: Kimberly M
authors.initials: KM
authors.lastname: Weber
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Elizabeth H
authors.initials: EH
authors.lastname: LeMasters
authors.affiliation: Centre for Communicable Diseases, The International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.
authors.forename: Sharifa
authors.initials: S
authors.lastname: Nasreen
authors.affiliation: Chembio Diagnostic Systems, Inc, Medford, NY, USA.
authors.forename: Javan
authors.initials: J
authors.lastname: Esfandiari
authors.affiliation: Chembio Diagnostic Systems, Inc, Medford, NY, USA.
authors.forename: Angelo H
authors.initials: AH
authors.lastname: Gunasekera
authors.affiliation: Department of Microbiology and Immunology, Emory University, Atlanta, GA, USA.
authors.forename: Megan
authors.initials: M
authors.lastname: McCausland
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.|Centre for Communicable Diseases, The International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b), Dhaka, Bangladesh.
authors.forename: Katharine
authors.initials: K
authors.lastname: Sturm-Ramirez
authors.affiliation: Department of Microbiology and Immunology, Emory University, Atlanta, GA, USA.
authors.forename: Jens
authors.initials: J
authors.lastname: Wrammert
authors.affiliation: Battelle Memorial Institute, Columbus, OH, USA.
authors.forename: Sean
authors.initials: S
authors.lastname: Gregory
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Vic
authors.initials: V
authors.lastname: Veguilla
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: James
authors.initials: J
authors.lastname: Stevens
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Miller
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Jacqueline M
authors.initials: JM
authors.lastname: Katz
authors.affiliation: Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA.
authors.forename: Min Z
authors.initials: MZ
authors.lastname: Levine
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D001459:Bangladesh; D001715:Bird Diseases / Q000097:blood / Q000821:virology; D001717:Birds; D003429:Cross Reactions; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D009980:Influenza A virus / Q000145:classification / Q000276:immunology* / Q000302:isolation & purification; D007251:Influenza, Human / Q000097:blood* / Q000821:virology; D013045:Species Specificity
chemical_list: D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus
keywords: 
  MAGPIX
  ; Chembio Dual Path Platform; antibody; hemagglutinin; influenza