Transient Expression of Zika NS2B Protein Antigen in Nicotiana benthamiana and Use for Arboviruses Diagnosis.

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 Nicotiana benthamiana 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.