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. 2024 Dec;96(12):e70100.
doi: 10.1002/jmv.70100.

Serotype-Specific Dengue Virus IgG Assay Using EDIII-Based Recombinant Proteins and Its Application in an Endemic Population in Northeast Brazil

Samuel Santos Pereira  1 , Rafael Rahal Guaragna Machado  2 , Jéssica Pires Farias  1 , Robert Adreata-Santos  1 , Mônica Josiane Rodrigues-Jesus  1 , Mayanna Moreira Costa Fogaça  3 , Milena Silva Souza  3 , Eduardo Gimenes Martins  1 , Camila Malta Romano  4 , Edison Luiz Durigon  2 , Jaime Henrique Amorim  3 , Luís Carlos de Souza Ferreira  1
Affiliations

Serotype-Specific Dengue Virus IgG Assay Using EDIII-Based Recombinant Proteins and Its Application in an Endemic Population in Northeast Brazil

Samuel Santos Pereira et al. J Med Virol. 2024 Dec.

Abstract

Dengue is the most prevalent arboviral disease globally, with Brazil currently experiencing a significant rise in cases. Dengue virus (DENV) typically co-circulates with other clinically and antigenically similar flaviviruses, such as Zika virus (ZIKV). The clinical diagnosis is difficult and accurate serological analysis represents an unmet challenge. Traditionally, serological analysis of DENV infection focuses on the acute phase via detection of NS1 or IgM. Developing IgG DENV-specific assays has been defiant due to the co-circulation of four antigenically distinct serotypes and a strong cross-reactivity with other arboviruses, particularly with ZIKV. The goal of this study was to produce recombinant domain III of the Envelope (EDIII) proteins of each DENV serotype and ZIKV and evaluate the ability to detect specific IgG antibodies. The antigens were tested on the ELISA platform to measure DENV-specific IgG in mice and patients infected with ZIKV and DENV. The assay differentiated serotype-specific IgG responses in susceptible mice (AG129) experimentally infected with DENV or ZIKV. In addition, the test demonstrated a robust performance achieving 87.8% sensitivity and 91.4% specificity when tested against 648 well-characterized sera collected from humans infected with DENV and/or ZIKV. The test was further applied to serum samples collected from 318 healthy individuals from an endemic region in Northwest/Central Brazil, without a previous diagnosis of DENV, and revealed that 65% of the samples reacted with at least one DENV serotype antigen, including 123 monotypic samples (88 for DENV-1) and 90 samples reacting with multiple antigens. Collectively, these results indicate that the IgG DENV-EDIII ELISA is a valuable tool for assessing the serological status of populations in endemic areas, particularly in regions where other flaviviruses, particularly ZIKV, co-circulate and offer support to establish public health policies against the disease.

Keywords: DENV; EDIII; ELISA; ZIKV; serological tests.

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