A multiplex serological assay for quantification of arbovirus antibody kinetics and neutralizing antibodies

Arboviruses such as dengue virus (DENV) and chikungunya virus (CHIKV) pose major global health threats. To support serological surveillance and study cross-reactivity, an in-house multiplex bead-based immunoassay was developed to measure IgG responses against a wide panel of orthoflavivirus and alphavirus antigens. Samples from Senegal, France, the Philippines, and Bangladesh enabled analyses to be carried out in endemic and non-endemic regions. IgG kinetics following DENV1 and CHIKV infections revealed virus-specific temporal profiles and identified antigens, such as CHIKV E2 and VLP, as well as DENV1 EDIII and SNAP-tagged EDIII, as promising markers for serosurveillance. A two-step Random Forest model was used to classify PRNT-positive samples and predict PRNT titers. CHIKV showed a strong correlation between IgG and PRNT titers (R² = 0.71), while DENV1 showed weaker performance (R² = 0.42). These results support the use of multiplex serology for arbovirus surveillance and highlight the limitations of using binding antibodies to predict neutralisation, particularly for orthoflaviviruses.