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

Young Chan Kim,Martha Eva Viveros-Sandoval,Carlos Alonso Domínguez-Alemán,Maria Antonieta Mar,Nallely Garcia-Larragoiti,César López-Camacho,Arturo Reyes-Sandoval,Alan Cano-Mendez,Héctor Vivanco-Cid,Karina Guadalupe Hernandez-Flores

doi:10.3390/v11050407

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.

Highlights

Chikungunya virus (CHIKV) is a re-emerging arbovirus that belongs to the Alphavirus genus of the Togaviridae family [1,2,3]
Seven out samples (10.3%) tested positive on reverse transcription-polymerase chain reaction (RT-PCR) confirming the acute CHIKV infection
407confirmed CHIKV patients have high mean titres of 3.4 and 2.7 on IgG and IgM ELISA, showed that respectively (Figure 3A right and Figure 3B right). These results indicate that our purified CHIKV E2 recombinant protein protein can can reliably reliably detect detect anti-CHIKV

Summary

Introduction

Chikungunya virus (CHIKV) is a re-emerging arbovirus that belongs to the Alphavirus genus of the Togaviridae family [1,2,3]. CHIKV infections are characterized by an acute onset of fever associated with myalgia, headache, and severe debilitating arthropathy [3]. Vaccines or specific anti-viral therapies for CHIKV are not yet available; rapid and simple methodologies to detect CHIKV infections are important for effective patient management and the control of future epidemics [12,14]. The primary laboratory test to diagnose CHIKV infection relies on the specific detection of CHIKV genomic sequences in serum collected up to five to six days after the onset of fever [15,16,17]. Several commercial and in-house CHIKV serological assays that became available are based on whole virus antigens, and those reports indicate a variation of sensitivities and specificities [18,19,20,21,22]

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