A Multiplex Serological Assay for the Detection of Antibody Responses to Arboviruses

Arthropod borne viruses (arboviruses) are a group of mosquito-borne pathogens that are globally distributed. Since the 2016 outbreak of Zika virus (ZIKV) in the Americas that brought these infections to public awareness, arboviruses continue to impose a considerable public health burden. Their emergence in previously non-endemic regions is a cause of public health concern. The surveillance and control of arboviral infections is a current priority for the World Health Organization (WHO), which launched a global strategic plan to fight arboviral diseases in 2024. The multiplex® technology has revolutionized the field of serological surveillance, allowing simultaneous measurement of multiple antibodies while requiring minimum sample volume. Building on expertise in the development of multiplex serological tests, a bead-based multiplex assay for the detection of antibodies to arbovirus antigens was developed. The assay comprises multiple antigens specific to common arboviruses, including dengue (DENV), yellow fever (YFV), Zika, and chikungunya (CHIKV) viruses. A semi-automated method was used to covalently bind antigens to microsphere beads. This new protocol is highly effective, enabling the coupling of up to 96 antigens in a single experiment while ensuring reproducible results. Median Fluorescence Intensity (MFI) data were acquired using a user-friendly fluorescence imaging system.When applied to human serum samples from endemic countries, we anticipate that this assay will constitute a powerful tool for sero-surveillance efforts.