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Type: Journal article
Title: Recombinant M2e protein-based ELISA: A novel and inexpensive approach for differentiating avian influenza infected chickens from vaccinated ones
Author: Hemmatzadeh, F.
Sumarningsih, S.
Tarigan, S.
Indriani, R.
Dharmayanti, N.
Ebrahimie, E.
Igniatovic, J.
Citation: PLoS One, 2013; 8(2):1-9
Publisher: Public Library of Science
Issue Date: 2013
ISSN: 1932-6203
Statement of
Farhid Hemmatzadeh, Sumarningsih Sumarningsih, Simson Tarigan, Risa Indriani, N.L.P. Indi Dharmayanti, Esmaeil Ebrahimie, Jagoda Igniatovic
Abstract: Available avian influenza (AIV) serological diagnostic tests cannot distinguish vaccinated from naturally infected birds. Differentiation of vaccinated from infected animals (DIVA) is currently advocated as a means of achieving the full control of H5N1. In this study, for the first time, recombinant ectodomain of M2 protein (M2e) of avian influenza virus (H5N1 strain) was used for the DIVA serology test. M2e was cloned into pMAL-P4X vector and expressed in E. coli cells. We used Western blot to recognize the expressed M2e-MBP protein by chicken antisera produced against live H5N1 virus. Also, the specificity of M2e-MBP protein was compared to the M2e synthetic peptide via ELISA. In M2e-MBP ELISA, all sera raised against the live avian influenza viruses were positive for M2e antibodies, whereas sera from killed virus vaccination were negative. Furthermore, M2e-MBP ELISA of the field sera obtained from vaccinated and non-vaccinated chickens showed negative results, while challenged vaccinated chickens demonstrated strong positive reactions. H5N1-originated recombinant M2e protein induced broad-spectrum response and successfully reacted with antibodies against other AIV strains such as H5N2, H9N2, H7N7, and H11N6. The application of the recombinant protein instead of synthetic peptide has the advantages of continues access to an inexpensive reagent for performing a large scale screening. Moreover, recombinant proteins provide the possibility of testing the DIVA results with an additional technique such a Western blotting which is not possible in the case of synthetic proteins. All together, the results of the present investigation show that recombinant M2e-MBP can be used as a robust and inexpensive solution for DIVA test.
Keywords: Animals
Recombinant Fusion Proteins
Viral Matrix Proteins
Antibodies, Viral
Antigens, Viral
Enzyme-Linked Immunosorbent Assay
Sensitivity and Specificity
Gene Expression
Influenza A Virus, H5N1 Subtype
Influenza in Birds
Rights: © 2013 Hemmatzadeh 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.0056801
Appears in Collections:Agriculture, Food and Wine publications
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