A multiplex serological assay for quantification of arbovirus antibody kinetics and neutralizing antibodies
17 affiliations
- 1Infectious Disease Epidemiology and Analytics Unit, Department of Global Health, Institut Pasteur, Université Paris-Cité, INSERM U1347, Paris, France
- 2Environment and Infectious Risks Unit, Institut Pasteur, Paris, France
- 3Division of Infectious Diseases, Department of Medicine Solna, Karolinska Institutet, Stockholm, Sweden
- 4Department of Infectious Diseases, Södersjukhuset, Stockholm, Sweden
- 5Processus Infectieux en Milieu Insulaire Tropical (PIMIT), Université de La Réunion, INSERM U1187, CNRS 9192, IRD 249, Plateforme Technologique CYROI, Sainte-Clotilde, La Réunion, France
- 6Institut Pasteur, Université Paris Cité, Production and Purification of recombinant Proteins Platform, CNRS UMR3528, Paris, France
- 7International Vaccine Institute, Seoul, Republic of Korea
- 8Department of Virology, Walter Reed Army Institute of Research-Armed Forces Research Institute of Medical Sciences (AFRIMS), Bangkok, Thailand
- 9Philippines-AFRIMS Virology Research Unit, Cebu City, Philippines
- 10Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, United States of America
- 11International Centre for Diarrhoeal Disease Research, Bangladesh (icddr, b), Dhaka, Bangladesh
- 12Department of Pathology and Center for Vector-Borne and Zoonotic Diseases, University of Texas, Medical Branch, Galveston, Texas, United States of America
- 13Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas, United States of America
- 14MRC Centre for Global Infectious Disease Analysis, School of Public Health, Imperial College London, London, United Kingdom
- 15Department of Infection Biology, Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom
- 16Institut Pasteur de Dakar, Dakar, Senegal
- 17Department of Genetics, University of Cambridge, Cambridge, United Kingdom
Abstract
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. Author summary: Arboviruses such as dengue and chikungunya are transmitted by mosquitoes and affect millions of people worldwide. In many regions, several viruses circulate at the same time. Detecting past infections using blood tests is essential for understanding how widely these viruses spread, but this is complicated because antibodies against related viruses often cross-react. In this study, we developed a high-throughput blood test that can measure antibodies against many mosquito-borne viruses simultaneously using a very small amount of blood. We applied this approach to samples collected in four countries with different patterns of arbovirus circulation: Senegal, Bangladesh, the Philippines, and France. By analysing antibody responses after confirmed dengue and chikungunya infections, we identified specific viral components that help distinguish true infections from cross-reactive responses. We used machine-learning methods to test whether antibody measurements could predict how antibodies in blood can neutralize viruses. This approach worked well for chikungunya but was less accurate for dengue, reflecting the complex immunity to closely related viruses. Overall, our results show that multiplex antibody testing can improve arbovirus surveillance, especially where several viruses are circulating at the same time.
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BibTeX
@article{garcia2026multiplex,
title = {{A multiplex serological assay for quantification of arbovirus antibody kinetics and neutralizing antibodies}},
author = {Laura Garcia and Victor Yman and Marie-Fabrice Gasasira and Françoise Donnadieu and Jason Rosado and Lucie Cappuccio and Damien Hoinard and Philippe Desprès and Stéphane Petres and Jean-Claude Manuguerra and In-Kyu Yoon and Taweewan Hunsawong and Maria Theresa P. Alera and Emily S. Gurley and Nikos Vasilakis and Scott C. Weaver and Ziaur Rahman and Ilaria Dorigatti and Chris Drakeley and Inès Vigan-Womas and Makhtar Niang and Henrik Salje and Jessica Vanhomwegen and Michael White},
journal = {PLOS Neglected Tropical Diseases},
year = {2026},
month = sep,
doi = {10.1371/journal.pntd.0013704},
url = {https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0013704},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - A multiplex serological assay for quantification of arbovirus antibody kinetics and neutralizing antibodies
AU - Laura Garcia
AU - Victor Yman
AU - Marie-Fabrice Gasasira
AU - Françoise Donnadieu
AU - Jason Rosado
AU - Lucie Cappuccio
AU - Damien Hoinard
AU - Philippe Desprès
AU - Stéphane Petres
AU - Jean-Claude Manuguerra
AU - In-Kyu Yoon
AU - Taweewan Hunsawong
AU - Maria Theresa P. Alera
AU - Emily S. Gurley
AU - Nikos Vasilakis
AU - Scott C. Weaver
AU - Ziaur Rahman
AU - Ilaria Dorigatti
AU - Chris Drakeley
AU - Inès Vigan-Womas
AU - Makhtar Niang
AU - Henrik Salje
AU - Jessica Vanhomwegen
AU - Michael White
PY - 2026
DA - 2026/09/23
JO - PLOS Neglected Tropical Diseases
DO - 10.1371/journal.pntd.0013704
UR - https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0013704
AB - 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. Author summary: Arboviruses such as dengue and chikungunya are transmitted by mosquitoes and affect millions of people worldwide. In many regions, several viruses circulate at the same time. Detecting past infections using blood tests is essential for understanding how widely these viruses spread, but this is complicated because antibodies against related viruses often cross-react. In this study, we developed a high-throughput blood test that can measure antibodies against many mosquito-borne viruses simultaneously using a very small amount of blood. We applied this approach to samples collected in four countries with different patterns of arbovirus circulation: Senegal, Bangladesh, the Philippines, and France. By analysing antibody responses after confirmed dengue and chikungunya infections, we identified specific viral components that help distinguish true infections from cross-reactive responses. We used machine-learning methods to test whether antibody measurements could predict how antibodies in blood can neutralize viruses. This approach worked well for chikungunya but was less accurate for dengue, reflecting the complex immunity to closely related viruses. Overall, our results show that multiplex antibody testing can improve arbovirus surveillance, especially where several viruses are circulating at the same time.
SN - 1935-2735
ER -