Pereira Samuel Santos, Machado Rafael Rahal Guaragna, Farias Jéssica Pires, Adreata-Santos Robert, Rodrigues-Jesus Mônica Josiane, Fogaça Mayanna Moreira Costa, Souza Milena Silva, Martins Eduardo Gimenes, Romano Camila Malta, Durigon Edison Luiz, Amorim Jaime Henrique, Ferreira Luís Carlos de Souza
Laboratory of Vaccine Development, Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
Laboratory of Clinical and Molecular Virology, Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
J Med Virol. 2024 Dec;96(12):e70100. doi: 10.1002/jmv.70100.
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.
登革热是全球最普遍的虫媒病毒病，巴西目前的病例数正在显著上升。登革热病毒（DENV）通常与其他临床和抗原相似的黄病毒共同传播，例如寨卡病毒（ZIKV）。临床诊断困难，准确的血清学分析是一项尚未解决的挑战。传统上，DENV感染的血清学分析通过检测NS1或IgM来关注急性期。由于四种抗原性不同的血清型共同传播以及与其他虫媒病毒，特别是与ZIKV有很强的交叉反应性，开发针对DENV的IgG特异性检测方法一直具有挑战性。本研究的目的是生产每种DENV血清型和ZIKV的包膜蛋白（EDIII）的重组结构域III，并评估检测特异性IgG抗体的能力。在ELISA平台上对这些抗原进行测试，以测量感染ZIKV和DENV的小鼠和患者体内的DENV特异性IgG。该检测方法能够区分实验感染DENV或ZIKV的易感小鼠（AG129）中的血清型特异性IgG反应。此外，在针对从感染DENV和/或ZIKV的人类收集的648份特征明确的血清进行测试时，该检测方法表现出色，灵敏度达到87.8%，特异性达到91.4%。该检测方法进一步应用于从巴西西北部/中部一个流行地区收集的318名健康个体的血清样本，这些个体之前未被诊断为DENV感染，结果显示65%的样本与至少一种DENV血清型抗原发生反应，包括123份单型样本（88份针对DENV-1）和90份与多种抗原发生反应的样本。总体而言，这些结果表明，IgG DENV-EDIII ELISA是评估流行地区人群血清学状况的有价值工具，特别是在其他黄病毒，尤其是ZIKV共同传播的地区，有助于制定针对该疾病的公共卫生政策。
