Pereira Samuel Santos, Andreata-Santos Robert, de Castro-Amarante Maria Fernanda, Venceslau-Carvalho Aléxia Adrianne, Sales Natiely Silva, Silva Mariângela de Oliveira, Alves Rúbens Prince Dos Santos, Jungmann Patrícia, Ferreira Luís Carlos de Souza
Laboratory of Vaccine Development, Institute of Biomedical Sciences, Department of Microbiology, University of São Paulo, São Paulo 05508-000, Brazil.
Institut Pasteur de São Paulo, São Paulo 05508-020, Brazil.
Viruses. 2023 Sep 16;15(9):1936. doi: 10.3390/v15091936.
Dengue is an infectious disease of global health concern that continues to require surveillance. Serological testing has been used to investigate dengue-infected patients, but specificity is affected by the co-circulation of ZIKA virus (ZIKV), which shares extensive antigen similarities. The goal of this study was the development of a specific dengue virus (DENV) IgG ELISA based on a multi-epitope NS1-based antigen for antibody detection. The multi-epitope protein (T-ΔNS1), derived from a fragment of the NS1-protein of the four DENV serotypes, was expressed in and purified via affinity chromatography. The antigenicity and specificity were evaluated with sera of mice infected with DENV-1-4 or ZIKV or after immunization with the recombinant ΔNS1 proteins. The performance of the T-ΔNS1-based IgG ELISA was also determined with human serum samples. The results demonstrate that the DENV T-ΔNS1 was specifically recognized by the serum IgG of dengue-infected mice or humans but showed no or reduced reactivity with ZIKV-infected subjects. Based on the available set of clinical samples, the ELISA based on the DENV T-ΔNS1 achieved 77.42% sensitivity and 88.57% specificity. The results indicate that the T-ΔNS1 antigen is a promising candidate for the development of specific serological analysis.
登革热是一种全球关注的传染病，仍需进行监测。血清学检测已被用于调查登革热感染患者，但特异性受到寨卡病毒（ZIKV）共同传播的影响，寨卡病毒与登革热病毒有广泛的抗原相似性。本研究的目的是开发一种基于多表位NS1抗原的特异性登革热病毒（DENV）IgG ELISA，用于抗体检测。从四种登革热病毒血清型的NS1蛋白片段衍生而来的多表位蛋白（T-ΔNS1）在大肠杆菌中表达，并通过亲和层析进行纯化。用感染DENV-1-4或ZIKV的小鼠血清或用重组ΔNS1蛋白免疫后的血清评估其抗原性和特异性。还用人类血清样本测定了基于T-ΔNS1的IgG ELISA的性能。结果表明，DENV T-ΔNS1被登革热感染小鼠或人类的血清IgG特异性识别，但与寨卡病毒感染受试者的反应性无或降低。基于现有的临床样本集，基于DENV T-ΔNS1的ELISA灵敏度为77.42%，特异性为88.57%。结果表明，T-ΔNS1抗原是开发特异性血清学分析的有前途的候选物。
