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DENGUE VIRUS TETRA-EPITOPE PEPTIDE EXPRESSED IN LETTUCE CHLOROPLASTS FOR POTENTIAL USE IN DENGUE DIAGNOSIS
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Universidade de Brasília. Departamento de Patologia Molecular. Brasília, DF, Brasil.
Embrapa Recursos Genéticos e Biotecnologia. Brasília, DF, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivirus. Rio de Janeiro, RJ, Brasil.
Universidade Católica de Brasília. Centro de Análises Proteomicas e Bioquímicas. Pós-Graduação em Ciências Genômicas e Biotecnologia. Brasília, DF, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivirus. Rio de Janeiro, RJ, Brasil.
Universidade Católica de Brasília. Centro de Análises Proteomicas e Bioquímicas. Pós-Graduação em Ciências Genômicas e Biotecnologia. Brasília, DF, Brasil.
Universidade de Brasília. Departamento de Patologia Molecular. Brasília, DF, Brasil.
Embrapa Recursos Genéticos e Biotecnologia. Brasília, DF, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivirus. Rio de Janeiro, RJ, Brasil.
Universidade Católica de Brasília. Centro de Análises Proteomicas e Bioquímicas. Pós-Graduação em Ciências Genômicas e Biotecnologia. Brasília, DF, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Flavivirus. Rio de Janeiro, RJ, Brasil.
Universidade Católica de Brasília. Centro de Análises Proteomicas e Bioquímicas. Pós-Graduação em Ciências Genômicas e Biotecnologia. Brasília, DF, Brasil.
Universidade de Brasília. Departamento de Patologia Molecular. Brasília, DF, Brasil.
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 feverpositive
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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