Development of an E2 ELISA Methodology to Assess Chikungunya Seroprevalence in Patients from an Endemic Region of Mexico

Abstract

Chikungunya fever is a debilitating disease caused by Chikungunya virus (CHIKV) that can result in long-lasting arthralgias. The early diagnosis of CHIKV relies on PCR during the acute infection phase to allow differential diagnosis with other co-circulating arboviruses such as dengue and Zika. Alternatively, serology can support diagnosis and provide epidemiological information on current and past outbreaks. Many commercial serological ELISA assays are based on the inactivated whole CHIKV, but their sensitivity and specificity show great variability. We produced recombinant CHIKV E2 that is suitable for ELISA assays, which was used for the serodiagnosis of CHIKV infections occurring in an arbovirus endemic Mexican region within Michoacán state. A cross-sectional study was conducted in 2016–2017; sera was obtained from 15 healthy donors and 68 patients presenting undifferentiated febrile illness. Serum samples were screened by RT-PCR and by our in-house ELISA assay. Our results indicate that IgM and IgG anti-CHIKV E2 antibodies were detected with our ELISA assay with higher sensitivity than a commercially available CHIKV ELISA kit. Our simple and sensitive ELISA assay for the serodiagnosis of CHIKV infections can be applied to population-based seroprevalence surveys and has potential for monitoring vaccine immunogenicity in CHIKV vaccine clinical trials.

Sero-prevalence of arthropod-borne viral infections among Lukanga swamp residents in Zambia

“…Biological samples were collected prior to vaccination and at specified time points, that is baseline, day 28,6,12,18,24,30,36,42, and 48 months. In order to test for antibodies against circulating arboviruses, we developed an enzyme-linked immunosorbent assay (ELISA) based on antigens specific to CHIKV and MAYV (E2), and DENV and ZIKV (NS1) arboviruses, which have been used to study patients from endemic regions of Mexico previously [34]. All serum samples collected at baseline and saliva samples from the same individuals collected a year later during follow up were subjected to the ELISA assay.…”

“…The ELISA assay on both baseline serum samples and saliva colleted at 1 year was conducted as described previously [34,35]. Briefly, plates were coated with 2 μg/mL of antigen diluted in PBS (coating buffer).…”

Chikungunya E2 Protein Produced in E. coli and HEK293-T Cells—Comparison of Their Performances in ELISA

“…Conversely, mammalian cells have the ability to express complex recombinant proteins with proper folding and glycosylation. A drawback, however, is the introduction of the exogenous gene to cells may be time-consuming and more complex, increasing the overall cost of protein production in comparison to prokaryotic expression systems [ 23 ].…”

Chikungunya Virus and (Re-) Emerging Alphaviruses

“…Thus, phylogenetic and epidemiological studies, clinical diagnostics, vaccine/antiviral drug development, basic research including virus-vector interaction, transmission, viral immunity, and pathogenesis are still necessary for alphaviral disease prevention. In this special issue of “Chikungunya Virus and (Re-) Emerging Alphaviruses”, we solicit 10 research articles and six review articles covering the development of vaccines and antivirals [1,2,3,4], pathogenesis/immunity [5,6,7,8], viral evolution [9], development of research/diagnostic tools [10,11], vector-virus interaction [12,13], as well as mechanisms of transmission [14].…”

“…Accurate diagnosis of viral infection is essential for appropriate medical treatment. Kim et al have developed a simple and sensitive enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of CHIKV infections [11]. This assay could be applied to population-based seroprevalence survey and evaluation of vaccination efficacy in clinical trials.…”