Expression and Evaluation of Chikungunya Virus E1 and E2 Envelope Proteins for Serodiagnosis of Chikungunya Virus Infection

Abstract

PurposeChikungunya virus (CHIKV) causes endemic or epidemic outbreaks of CHIKV fever, which is a mosquitoe-transmitted viral disease in Africa, India, South-East Asia, and recently Southern Europe. Currently, serological diagnostic tests such as hemagglutination inhibition test (HI test), in-house IgM capture enzyme-linked immunosorbent assays (ELISA), and indirect immunofluorescence test were used for diagnosis of chikungunya fever, which are based on whole virus antigens.Materials and MethodsCHIKV E1, and E2 envelope proteins for the CHIKV-specific serodiagnostic reagents for chikungunya fever were expressed in baculovirus expression system. The seroreactivity of recombinant CHIKV E1 and E2 envelope proteins were evaluated using sera panels of patients from Laboratoire Marcel Merieux by indirect IgM capture ELISA.ResultsThe recombinant CHIKV E1 and E2 envelope protein showed sensitivity of 77.5% and 90%, respectively. The specificities of both CHIKV E1 and E2 envelope proteins were 100%.ConclusionThe recombinant CHIKV E1 and E2 envelope proteins could be a useful diagnostic reagent for CHIKV infection.

Analysis of antibody response (IgM, IgG, IgG3) to Chikungunya virus using panel of peptides derived from envelope protein for serodiagnosis

“…Immunoassays showed significantly high titer IgM, IgG, IgG3 antibodies and none of the peptides showed cross reactivity with Dengue positive sera. Thus the data suggest that the antibody response during early Chikungunya infection is mainly dominated by E2 peptides, a finding in accordance with other reports [34]. On the basis of present analysis, we can conclude that a combination of highly specific and sensitive peptides derived from E2 antigen can form the basis for designing a peptide-based diagnostic assay.…”

Diagnostic potential of monoclonal antibodies against the capsid protein of chikungunya virus for detection of recent infection

“…The E protein or the E protein gene of CHIKV has always been considered a major target for the diagnosis of CHIKV infection in serological and molecular assays [39, 41,42,51,52], but recently, the C protein has also been explored as a target for CHIKV diagnosis in various assays. Reports indicate 85 % sensitivity and 100 % specificity [39] and 97 % accordance of indirect IgM detection ELISA using recombinant C protein when compared to a commercial kit [40]. Cho et al reported that using recombinant C protein in ICA gave 87.5 % sensitivity with 100 % specificity [38].…”

“…These antigens have been obtained from clinical specimens or cell culture. Recombinant C-, E1-and E2-protein-based indirect IgM ELISA and immunochromatography assays (ICA) exhibited sensitivity in the range of 77.5 %-100 % with 95 %-100 % specificity [38][39][40]. MAbs to E2 protein successfully detected CHIKV derived from mosquito and human cells in antigen-capture ELISA [41].…”

Arbovirus vaccines; opportunities for the baculovirus-insect cell expression system

“…Several studies have shown that baculovirus-mediated expression of alphavirus structural proteins C, E3, E2, 6K and E1, results in correctly processed and mature (glyco)proteins in insect cells (Cho et al, 2008;Favre et al, 1993;Hodgson et al, 1999;Oker-Blom and Summers, 1989). Although the baculovirus expression system serves as an elegant platform for subunit vaccine development and available data are promising, this is still a fairly uncharted area and few studies have focused on the immunogenicity of alphavirus subunits produced in insect cells.…”