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Specific subtyping of influenza A virus using a recombinant hemagglutinin protein expressed in baculovirus

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Abstract

Influenza A viruses are subtyped according to antigen characterization of hemagglutinin (HA) and neuraminidase surface glycoproteins. The hemagglutination inhibition (HI) assay using reference antiserum is currently applied to serologic screening of subtype-specific antibodies in sera. The reference antiserum is made by injecting chickens with live or inactivated whole virus preparations. Nonspecific inhibitors of antisera prepared by the conventional method may affect the specificity of HI assay. In this study, highly pure recombinant proteins generated using baculovirus expression vector system based on full-length of HA (HAF) and antigenic region of HA1 genes of H9 subtype, and also inactivated whole virus were used to immunization of chickens. Measurable antibody titers were present for treated birds after 3 weeks and generally increased after each boost. The performance of the prepared antisera was evaluated by testing a panel of known standard strains of influenza virus representing five HA subtypes. Relative to the conventional method using whole virus immunization and recombinant HAF protein, the antiserum prepared by recombinant HA1 had a specificity of 100% for all tested subtypes. The antiserum prepared by expression of HA1 protein in baculovirus has the potential for rapid and specific HA subtyping of influenza viruses without producing antibodies specific to other viral proteins.

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Authors and Affiliations

  1. National Institute of Genetic Engineering and Biotechnology, P.O. Box 14155-6343, Tehran, Iran

    Shahla Shahsavandi, Ali-Hatef Salmanian & Seyed Ali Ghorashi

  2. Razi Vaccine and Serum Research Institute, P.O. Box 31975-148, Karaj, Iran

    Shahla Shahsavandi, Shahin Masoudi & Mohammad Majid Ebrahimi

  3. Pasteur Institute of Iran, P.O. Box 1316-943551, Tehran, Iran

    Fatemeh Fotouhi

Authors
  1. Shahla Shahsavandi
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  2. Ali-Hatef Salmanian
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  3. Seyed Ali Ghorashi
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  4. Shahin Masoudi
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  5. Fatemeh Fotouhi
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  6. Mohammad Majid Ebrahimi
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Correspondence to Ali-Hatef Salmanian.

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Shahsavandi, S., Salmanian, AH., Ghorashi, S.A. et al. Specific subtyping of influenza A virus using a recombinant hemagglutinin protein expressed in baculovirus. Mol Biol Rep 38, 3293–3298 (2011). https://doi.org/10.1007/s11033-010-0434-2

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  • DOI: https://doi.org/10.1007/s11033-010-0434-2

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