Development of an enzyme-linked immunosorbent assay using recombinant protein antigen for the diagnosis of Chikungunya virus
Flávia Fonseca Bagno,
Lara Carvalho Godoi,
Natalia Salazar,
Glauco de Carvalho Pereira,
Maria Marta Figueiredo,
Flávio Guimarães da Fonseca
Affiliations
Flávia Fonseca Bagno
Departamento de Microbiologia, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil; Centro de Tecnologia de Vacinas/UFMG, Belo Horizonte, Brazil
Lara Carvalho Godoi
Centro de Tecnologia de Vacinas/UFMG, Belo Horizonte, Brazil; Colégio Técnico da Universidade Federal de Minas Gerais, Belo Horizonte, Brazil
Natalia Salazar
Centro de Tecnologia de Vacinas/UFMG, Belo Horizonte, Brazil
Glauco de Carvalho Pereira
Fundação Ezequiel Dias (FUNED), Belo Horizonte, Brazil
Maria Marta Figueiredo
Centro de Tecnologia de Vacinas/UFMG, Belo Horizonte, Brazil
Flávio Guimarães da Fonseca
Departamento de Microbiologia, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil; Centro de Tecnologia de Vacinas/UFMG, Belo Horizonte, Brazil; Corresponding author. Departamento de Microbiologia, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
We describe here the development of an in-house enzyme linked immunosorbent assay (ELISA) for the diagnostic of Chikungunya virus (CHIKV) infections using a recombinant protein from CHIKV. The recombinant protein gene was designed based on 154 sequences and we used computational methods to predict its structure and antigenic potential. To confirm predictions, the gene coding for the recombinant CHIKV protein (rCHIKVp) was synthetized and expressed in prokaryotic system. Subsequently, the protein was purified by affinity chromatography and used as antigen in an indirect ELISA. We present data regarding the optimization of the recombinant antigen production and preparation of the ELISA to detect IgG against CHIKV in human sera. Keywords: Chikungunya virus, Diagnostics, ELISA, E2 Envelope protein, Recombinant protein
