Dengue virus tetra-epitope peptide expressed in lettuce chloroplasts for potential use in dengue diagnosis
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BIOTECHNOLOGICAL PRODUCTS AND PROCESS ENGINEERING
Dengue virus tetra-epitope peptide expressed in lettuce chloroplasts for potential use in dengue diagnosis Franciele Roberta Maldaner & Francisco José Lima Aragão & Flávia Barreto dos Santos & Octavio Luiz Franco & Monique da Rocha Queiroz Lima & Renato de Oliveira Resende & Raquel Medeiros Vasques & Tatsuya Nagata
Received: 24 January 2013 / Revised: 4 April 2013 / Accepted: 8 April 2013 / Published online: 25 April 2013 # Springer-Verlag Berlin Heidelberg 2013
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, Electronic supplementary material The online version of this article (doi:10.1007/s00253-013-4918-6) contains supplementary material, which is available to authorized users. F. R. Maldaner : T. Nagata Departamento de Patologia Molecular, Universidade de Brasília, 70910-900 Brasília, Federal District, Brazil F. J. L. Aragão (*) Embrapa Recursos Genéticos e Biotecnologia, PqEB W5 Norte, 70770-900 Brasília, Federal District, Brazil e-mail: [email protected] F. B. dos Santos : M. da Rocha Queiroz Lima Laboratório de Flavivirus, Instituto Oswaldo Cruz, 21040-360 Rio de Janeiro, Rio de Janeiro, Brazil O. L. Franco : R. M. Vasques 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 R. de Oliveira Resende : T. Nagata (*) Departamento de Biologia Celular, Universidade de Brasília, 70910-970 Brasília, Federal District, Brazil e-mail: [email protected]
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. Keywords Dengue vir
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