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Dengue virus tetra-epitope peptide expressed in lettuce chloroplasts for potential use in dengue diagnosis

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Abstract

Dengue virus causes about 100 million cases of dengue disease per year in the world. Laboratory diagnosis is done mainly by serological techniques, which in many cases use crude virus extracts that may cause cross-reactions to other flaviviruses. These undesirable cross-reactions can be reduced or eliminated by using recombinant proteins based on restricted epitopes. Aiming to decrease flaviviral cross-reactions and non-specific interactions in dengue serological assays, a plant expression system was chosen for recombinant antigen production as a reliable and inexpensive dengue diagnostic tool. In the present report, the lettuce plastid transformation system was applied to achieve efficient and stable tetra-epitope peptide antigen production, and its reactivity was evaluated. For this purpose, one putative epitope at positions 34 to 57 of E protein within the junction site of domains I and II of dengue virus (DENV) 1 to 4 serotypes linked by glycine linkers was expressed in lettuce chloroplasts. The potential immunoreactivity for the four DENV serotypes was evaluated using sera from patients of positive and negative dengue cases. Results indicated an overall sensitivity of 71.7 % and specificity of 100 %. No cross-reactions with the sera of yellow fever-positive or healthy individuals vaccinated against yellow fever were observed. This novel approach may provide an alternative system for the large-scale production of dengue recombinant antigens useful for serodiagnosis.

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Acknowledgments

We thank Dr. Hirosuke Kanamoto (Research Institute of Innovative Technology for the Earth, Japan) for providing plasmid pRL1001. We also thank to Dr. Rita Maria Nogueira (Flavivirus Laboratory, IOC/Fiocruz, Brazil) for providing the samples. The work was funded by the Fundação de Apoio à Pesquisa do Distrito Federal (Brazil) for a research grant and by the “Conselho Nacional de Desenvolvimento Científico e Tecnológico” (CNPq, Brazil) for the DSc scholarship to the first author.

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Authors and Affiliations

  1. Departamento de Patologia Molecular, Universidade de Brasília, 70910-900, Brasília, Federal District, Brazil

    Franciele Roberta Maldaner & Tatsuya Nagata

  2. Embrapa Recursos Genéticos e Biotecnologia, PqEB W5 Norte, 70770-900, Brasília, Federal District, Brazil

    Francisco José Lima Aragão

  3. Laboratório de Flavivirus, Instituto Oswaldo Cruz, 21040-360, Rio de Janeiro, Rio de Janeiro, Brazil

    Flávia Barreto dos Santos & Monique da Rocha Queiroz Lima

  4. Pós-Graduação em Ciências Genômicas e Biotecnologia, Centro de Análises Proteomicas e Bioquímicas, Universidade Católica de Brasília, SGAN Quadra 916, Módulo B, Av. W5 Norte, 70790-160, Brasília, Federal District, Brazil

    Octavio Luiz Franco & Raquel Medeiros Vasques

  5. Departamento de Biologia Celular, Universidade de Brasília, 70910-970, Brasília, Federal District, Brazil

    Renato de Oliveira Resende & Tatsuya Nagata

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  1. Franciele Roberta Maldaner
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  2. Francisco José Lima Aragão
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  8. Tatsuya Nagata
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Correspondence to Francisco José Lima Aragão or Tatsuya Nagata.

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Maldaner, F.R., Aragão, F.J.L., dos Santos, F.B. et al. Dengue virus tetra-epitope peptide expressed in lettuce chloroplasts for potential use in dengue diagnosis. Appl Microbiol Biotechnol 97, 5721–5729 (2013). https://doi.org/10.1007/s00253-013-4918-6

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  • DOI: https://doi.org/10.1007/s00253-013-4918-6

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