Development and assessment of a multiepitope synthetic antigen for the diagnosis of Dengue virus infection.

Autor:	da Silva IBN; Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica Leopoldo de Meis, Rio de Janeiro, RJ, Brazil., de Moraes Rodrigues J; Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica Leopoldo de Meis, Rio de Janeiro, RJ, Brazil., Batista RCG; Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica Leopoldo de Meis, Rio de Janeiro, RJ, Brazil., Gomes VDS; Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica Leopoldo de Meis, Rio de Janeiro, RJ, Brazil., Chacon CS; Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica Leopoldo de Meis, Rio de Janeiro, RJ, Brazil., Almeida MDS; Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica Leopoldo de Meis, Rio de Janeiro, RJ, Brazil; Universidade Federal do Rio de Janeiro, Centro Nacional de Biologia Estrutural e Bioimagem, Plataforma Avançada de Biomoléculas, Rio de Janeiro, RJ, Brazil., de Araujo TS; Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica Leopoldo de Meis, Rio de Janeiro, RJ, Brazil; Universidade Federal do Rio de Janeiro, Centro Nacional de Biologia Estrutural e Bioimagem, Plataforma Avançada de Biomoléculas, Rio de Janeiro, RJ, Brazil., Ortiz da Silva B; Universidade Federal do Rio de Janeiro, Núcleo de Enfrentamento e Estudos de Doenças Infecciosas Emergentes e Reemergentes (NEEDIER), Rio de Janeiro, RJ, Brazil., Castiñeiras TMPP; Universidade Federal do Rio de Janeiro, Núcleo de Enfrentamento e Estudos de Doenças Infecciosas Emergentes e Reemergentes (NEEDIER), Rio de Janeiro, RJ, Brazil; Universidade Federal do Rio de Janeiro, Faculdade de Medicina, Departamento de Doenças Infecciosas e Parasitárias, Rio de Janeiro, RJ, Brazil., Ferreira Junior ODC; Universidade Federal do Rio de Janeiro, Núcleo de Enfrentamento e Estudos de Doenças Infecciosas Emergentes e Reemergentes (NEEDIER), Rio de Janeiro, RJ, Brazil; Universidade Federal do Rio de Janeiro, Instituto de Biologia, Laboratório de Virologia Molecular, Rio de Janeiro, RJ, Brazil., Carneiro FA; Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica Leopoldo de Meis, Rio de Janeiro, RJ, Brazil; Universidade Federal do Rio de Janeiro, Núcleo de Pesquisa (Numpex-Bio), Campus Duque de Caxias Professor Geraldo Cidade, Duque de Caxias, RJ, Brazil., Montero-Lomeli M; Universidade Federal do Rio de Janeiro, Instituto de Bioquímica Médica Leopoldo de Meis, Rio de Janeiro, RJ, Brazil. Electronic address: montero@bioqmed.ufrj.br.
Jazyk:	angličtina
Zdroj:	The Brazilian journal of infectious diseases : an official publication of the Brazilian Society of Infectious Diseases [Braz J Infect Dis] 2024 May-Jun; Vol. 28 (3), pp. 103746. Date of Electronic Publication: 2024 May 01.
DOI:	10.1016/j.bjid.2024.103746
Abstrakt:	Immunodiagnostic tests for detecting dengue virus infections encounter challenges related to cross-reactivity with other related flaviviruses. Our research focuses on the development of a synthetic multiepitope antigen tailored for dengue immunodiagnostics. Selected dengue epitopes involved structural linearity and dissimilarity from the proteomes of Zika and Yellow fever viruses which served for computationally modeling the three-dimensional protein structure, resulting in the design of two proteins: rDME-C and rDME-BR. Both proteins consist of seven epitopes, separated by the GPGPG linker, and a carboxy-terminal 6 × -histidine tag. The molecular weights of the final proteins rDME-C and rDME-BR are 16.83 kDa and 16.80 kDa, respectively, both with an isoelectric point of 6.35. The distinguishing factor between the two proteins lies in the origin of their epitope sequences, where rDME-C is based on the reference dengue proteome, while rDME-BR utilizes sequences from prevalent Dengue genotypes in Brazil from 2008 to 2019. PyMol analysis revealed exposure of epitopes in the secondary structure. Successful expression of the antigens was achieved in soluble form and fluorescence experiments indicated a disordered structure. In subsequent testing, rDME-BR and rDME-C antigens were assessed using an indirect Elisa protocol against Dengue infected serum, previously examined with a commercial diagnostic test. Optimal concentrations for antigens were determined at 10 µg/mL for rDME-BR and 30 µg/mL for rDME-C, with serum dilutions ranging from 1:50 to 1:100. Both antigens effectively detected IgM and IgG antibodies in Dengue fever patients, with rDME-BR exhibiting higher sensitivity. Our in-house test showed a sensitivity of 77.3 % and 82.6 % and a specificity of 89.4 % and 71.4 % for rDME-C and rDEM-BR antigens. No cross-reactivity was observed with serum from Zika-infected mice but with COVID-19 serum samples. Our findings underscore the utility of synthetic biology in crafting Dengue-specific multiepitope proteins and hold promise for precise clinical diagnosis and monitoring responses to emerging Dengue vaccines. Competing Interests: Conflicts of interest The authors declare no conflicts of interest. (Copyright © 2024 Sociedade Brasileira de Infectologia. Published by Elsevier España, S.L.U. All rights reserved.)
Databáze:	MEDLINE
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