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Influenza Other Respir Viruses. 2017 Feb 16. doi: 10.1111/irv.12449. [Epub ahead of print]
Novel multiplex assay platforms to detect influenza A hemagglutinin subtype specific antibody responses for high-throughput and in-field applications.
Li ZN1, Trost JF1,2, Weber KM3, LeMasters EH1, Nasreen S4, Esfandiari J5, Gunasekera AH5, McCausland M2, Sturm-Ramirez K1,4, Wrammert J2, Gregory S3, Veguilla V1, Stevens J1, Miller JD1, Katz JM1, Levine MZ1.
Author information
Abstract
BACKGROUND:
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.
METHODS:
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(H1N1) and A(H3N2) were introduced into the platforms to reduce cross-reactivity.
RESULTS:
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% to 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% (6/6) for MAGPIX, 83% (5/6) for DPP; H5 specificity was 100% (15/15) and cross-reactivity against other subtype was 0% (0/6) for both platforms.
CONCLUSION:
MAGPIX and DPP platforms can be utilized for high-throughput and in-field detection of novel influenza virus infections. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
KEYWORDS:
MAGPIX ; Chembio DPP; antibody; hemagglutinin; influenza
PMID: 28207986 DOI: 10.1111/irv.12449
[PubMed - as supplied by publisher] Free full text
Novel multiplex assay platforms to detect influenza A hemagglutinin subtype specific antibody responses for high-throughput and in-field applications.
Li ZN1, Trost JF1,2, Weber KM3, LeMasters EH1, Nasreen S4, Esfandiari J5, Gunasekera AH5, McCausland M2, Sturm-Ramirez K1,4, Wrammert J2, Gregory S3, Veguilla V1, Stevens J1, Miller JD1, Katz JM1, Levine MZ1.
Author information
Abstract
BACKGROUND:
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.
METHODS:
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(H1N1) and A(H3N2) were introduced into the platforms to reduce cross-reactivity.
RESULTS:
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% to 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% (6/6) for MAGPIX, 83% (5/6) for DPP; H5 specificity was 100% (15/15) and cross-reactivity against other subtype was 0% (0/6) for both platforms.
CONCLUSION:
MAGPIX and DPP platforms can be utilized for high-throughput and in-field detection of novel influenza virus infections. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
KEYWORDS:
MAGPIX ; Chembio DPP; antibody; hemagglutinin; influenza
PMID: 28207986 DOI: 10.1111/irv.12449
[PubMed - as supplied by publisher] Free full text