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Abstract
Chikungunya (CHIKV), an arbovirus that belongs to the Alphavirus genus of the Togaviridae family, causes a disease characterized by acute onset of fever accompanied by arthralgia. CHIKV also has been associated with cases of meningoencephalitis (primarily in neonates), Guillain–Barré syndrome and hemorrhagic disease. The clinical similarities, cross-reactivity and cocirculation of arboviruses in Brazil have complicated their differentiation, highlighting the need for new diagnostic tools. A serologic test can be useful for acute detections as well as for surveillance and epidemiological studies. Point of care tests currently available to detect CHIKV infections have been associated with low accuracy. The aim of this work was to design a synthetic gene and generate a recombinant protein to be used as antigens in diagnostic assays. Computational methods were used to predict its structure and antigenic potential. To confirm predictions, the gene coding for the recombinant CHIKV protein (rCHIKV) was synthetized and the protein was purified by affinity chromatography. Antigenicity of the protein was initially confirmed by western-blot using sera from CHIKV infected mice. Additionally, the seroreactivity of r-CHIKV protein was evaluated using a panel of sera samples from human patients, CHIKV seropositive or not, by indirect IgG ELISA. The r-CHIKV protein showed sensitivity of 95 % and specificity of 96 %. No cross-reactivity was found against sera of Zika and Dengue positive patients. These results indicate that this proteins maybe useful antigen to detect CHIKV infections in ELISA assays. Our next step includes the development of a rapid test kit.
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