Herazo Mario A M, Dantas Daylana R S, Silva Bruno B, Costa Helen P S, Santos Eduarda N F N, Moura Luiz F W G, Neto João X S, Van Tilburg Maurício F, Florean Eridan O P T, Moura Arlindo A, Guedes Maria I F
Laboratory of Biotechnology and Molecular Biology, Health Sciences Center, State University of Ceara, Fortaleza 60714-903, Brazil.
Federal University of Ceara, Fortaleza 60355-636, Brazil.
ACS Omega. 2025 Jan 8;10(2):2184-2196. doi: 10.1021/acsomega.4c08998. eCollection 2025 Jan 21.
Zika (ZIKV) and Dengue (DENV) viruses are clinically significant due to their severe neurological and hemorrhagic complications. Rapid diagnostics often rely on nonstructural proteins to generate specific antibodies. This study aimed to produce IgG antibodies from the recombinant ZIKV protein and plant-expressed NS2B protein for arbovirus detection in serum and urine samples. The NS2B protein was expressed in and purified chromatographically. Validation of recombinant NS2B as an antigen in indirect immunoassays demonstrated 95% sensitivity and 100% specificity in IgM/IgG ELISA tests, enabling effective detection of ZIKV and DENV. Notably, r-ZIKV-NS2B IgG identified positive ZIKV and DENV cases in urine but failed to detect negatives, suggesting limitations in specificity for urine diagnostics. Using urine as a diagnostic medium offers a less invasive and more practical approach, broadening the test applicability. This study utilized patient-derived positive urine samples and healthy samples spiked with an exogenous virus. Findings highlight the potential of the ZIKV-NS2B protein as a robust antigen for arbovirus diagnosis and demonstrate the viability of plant-based systems for antigen production, advancing diagnostics for neglected tropical diseases.
寨卡病毒（ZIKV）和登革热病毒（DENV）因其严重的神经和出血性并发症而具有临床重要性。快速诊断通常依赖非结构蛋白来产生特异性抗体。本研究旨在从重组寨卡病毒蛋白和植物表达的NS2B蛋白中产生IgG抗体，用于检测血清和尿液样本中的虫媒病毒。NS2B蛋白在[具体表达系统未给出]中表达并通过色谱法纯化。在间接免疫测定中，将重组NS2B作为抗原进行验证，结果显示在IgM/IgG ELISA试验中灵敏度为95%，特异性为100%，能够有效检测寨卡病毒和登革热病毒。值得注意的是，重组寨卡病毒NS2B IgG在尿液中能识别出寨卡病毒和登革热病毒阳性病例，但无法检测出阴性病例，这表明其在尿液诊断的特异性方面存在局限性。将尿液用作诊断介质提供了一种侵入性较小且更实用的方法，拓宽了检测的适用性。本研究使用了患者来源的阳性尿液样本和接种了外源病毒的健康样本。研究结果突出了寨卡病毒NS2B蛋白作为虫媒病毒诊断的强大抗原的潜力，并证明了基于植物的系统生产抗原的可行性，推动了被忽视热带病的诊断。
