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https://hdl.handle.net/2440/91169
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Type: | Journal article |
Title: | Multimeric recombinant M2e protein-based ELISA: A significant improvement in differentiating avian influenza infected chickens from vaccinated ones |
Author: | Hadifar, F. Ignjatovic, J. Tarigan, S. Indriani, R. Ebrahimie, E. Haliza Hasan, N. McWhorter, A. Putland, S. Ownagh, A. Hemmatzadeh, F. |
Citation: | PLoS One, 2014; 9(10):e108420-1-e108420-8 |
Publisher: | Public Library of Science |
Issue Date: | 2014 |
ISSN: | 1932-6203 1932-6203 |
Editor: | Tompkins, S.M. |
Statement of Responsibility: | Farshid Hadifar, Jagoda Ignjatovic, Simson Tarigan, Risa Indriani, Esmaeil Ebrahimie, Noor Haliza Hasan, Andrea McWhorter, Sophie Putland, Abdulghaffar Ownagh, Farhid Hemmatzadeh |
Abstract: | Killed avian influenza virus (AIV) vaccines have been used to control H5N1 infections in countries where the virus is endemic. Distinguishing vaccinated from naturally infected birds (DIVA) in such situations however, has become a major challenge. Recently, we introduced the recombinant ectodomain of the M2 protein (M2e) of H5N1 subtype as a novel tool for an ELISA based DIVA test. Despite being antigenic in natural infection the monomer form of the M2e used in ELISA had limited antigenicity and consequently poor diagnostic capability. To address this shortcoming, we evaluated the use of four tandem copies of M2e (tM2e) for increased efficiency of M2e antibody detection. The tM2e gene of H5N1 strain from Indonesia (A/Indonesia/CDC540/2006) was cloned into a pMAL- p4x expression vector and expressed in E.coli as a recombinant tM2e-MBP or M2e-MBP proteins. Both of these, M2e and tM2e antigens reacted with sera obtained from chickens following live H5N1 infection but not with sera from vaccinated birds. A significantly stronger M2e antibody reaction was observed with the tM2e compared to M2e antigen. Western blotting also supported the superiority of tM2e over M2e in detection of specific M2e antibodies against live H5N1 infection. Results from this study demonstrate that M2e tetramer is a better antigen than single M2e and could be more suitable for an ELISA based DIVA test. |
Keywords: | Animals Chickens Recombinant Fusion Proteins Viral Matrix Proteins Influenza Vaccines Antigens, Viral Blotting, Western Enzyme-Linked Immunosorbent Assay Cloning, Molecular Amino Acid Sequence Protein Structure, Quaternary Genetic Vectors Molecular Sequence Data Influenza A Virus, H5N1 Subtype Influenza in Birds Protein Multimerization |
Rights: | © 2014 Hadifar et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
DOI: | 10.1371/journal.pone.0108420 |
Published version: | http://dx.doi.org/10.1371/journal.pone.0108420 |
Appears in Collections: | Animal and Veterinary Sciences publications Aurora harvest 2 |
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hdl_91169.pdf | Published version | 1.11 MB | Adobe PDF | View/Open |
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