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Antigen production using heterologous expression of dengue virus-2 non-structural protein 1 (NS1) in Nicotiana tabacum (Havana) for immunodiagnostic purposes

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An Erratum to this article was published on 29 August 2016

Abstract

Key message

Expression of dengue-2 virus NS1 protein in Nicotiana tabacum plants for development of dengue immunodiagnostic kits.

Abstract

Dengue is one of the most important diseases caused by arboviruses in the world. A significant increase in its geographical distribution has been noticed over the last 20 years, with continuous transmission of several serotypes and emergence of the hemorrhagic fever in areas where the disease was previously not prevalent. Although the methodological processes for dengue diagnosis are in deep development and improvement, a limitation for the realization of dengue diagnostic tests is the difficulty of large-scale production of the antigen to be used in diagnostic tests. Due to this demand, the purpose of this study was to obtain the non-structural protein 1 (NS1) from dengue-2 serotype by heterologous expression in Nicotiana tabacum (Havana). After confirmation of the NS1 protein gene integration in the plant genome, the heterologous protein was characterized using SDS-PAGE and immunoblotting. In an immunoenzymatic test, the recombinant NS1 protein presents an antigen potential for development of dengue immunodiagnostic kits.

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Abbreviations

C:

Capsid protein

cDNA:

Complementar DNA

DENV:

Dengue virus

E:

Envelope protein

ELISA:

Enzyme Linked Immuno Sorbent Assay

IgG:

Immunoglobulin G

IgM:

Immunoglobulin M

kDa:

Kilodalton

NS1:

Non structural protein 1

NS2a:

Non-structural protein 2a

NS2b:

Non-structural protein 2b

NS3:

Non-structural protein 3

NS4a:

Non-structural protein 4a

NS4b:

Non-structural protein 4b

NS5:

Non-structural protein 5

PCR:

Polymerase chain reaction

PFU:

Plaque formation unit

prM:

Pre-membrane protein

ROC:

Receiver operating characteristic

RT-PCR:

Reverse transcription polymerase chain reaction

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Acknowledgments

This work was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brazil (Pronex Grant # 550165/2010-0 and INCT Dengue Grant # 471221/2008-2) and by Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG).

Conflict of interest

The authors declare that they have no competing interests.

Author information

Authors and Affiliations

  1. Laboratory of Molecular Immunovirology, Department of General Biology, Federal University of Viçosa, Av. PH Rolfs, s/n Campus Universitário, Viçosa, MG, CEP 36570-000, Brazil

    Marilane O. F. Amaro, Mariana F. Xisto, Ana Carolina F. Dias, Alice F. Versiani, Silvia A. Cardoso & Sérgio O. De Paula

  2. Department of Vegetal Biology, Federal University of Viçosa, Viçosa, MG, Brazil

    Alice F. Versiani & Wagner C. Otoni

  3. Department of Microbiology, Federal University of Viçosa, Viçosa, MG, Brazil

    Alice F. Versiani & Cynthia C. da Silva

Authors
  1. Marilane O. F. Amaro
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  2. Mariana F. Xisto
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  3. Ana Carolina F. Dias
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  4. Alice F. Versiani
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  5. Silvia A. Cardoso
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  6. Wagner C. Otoni
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  7. Cynthia C. da Silva
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  8. Sérgio O. De Paula
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Corresponding author

Correspondence to Sérgio O. De Paula.

Additional information

Communicated by B. Li.

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Amaro, M.O.F., Xisto, M.F., Dias, A.C.F. et al. Antigen production using heterologous expression of dengue virus-2 non-structural protein 1 (NS1) in Nicotiana tabacum (Havana) for immunodiagnostic purposes. Plant Cell Rep 34, 919–928 (2015). https://doi.org/10.1007/s00299-015-1753-5

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  • DOI: https://doi.org/10.1007/s00299-015-1753-5

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