One-Step Immunochromatographic Assay Enabling Differential Diagnosis of Dengue and Chikungunya

Dengue (DEN) and Chikungunya (CHIK) are both acute infectious diseases transmitted by Aedes albopictus, with similar clinical symptoms at the initial stage of infection. DEN virus (DENV) and CHIK virus (CHIKV) are the main causative pathogens of DEN and CHIK, respectively. This study aims to develop a rapid diagnostic immunochromatographic strip for differential diagnosis of DENV and CHIKV infections. A dual T line colloidal gold immunochromatographic assay (dual-T ICA) based on the double-antibody sandwich format was optimized and developed. Subsequently, a dual-T ICA strip (dual-T ICAS) was assembled and evaluated. A dual-T ICAS for the simultaneous detection of DENV NS1 protein and CHIKV E2 protein was successfully prepared. The assay achieved limits of detection of 0.2 ng/mL for DENV NS1 protein and 0.8 ng/mL for CHIKV E2 protein, demonstrating no cross-reactivity with tested heterologous viral pathogens as well as high precision and reproducibility. Results were clearly visible within 12 min. Comparative evaluation of 150 clinical samples revealed no statistically significant difference in diagnostic performance between the dual-T ICAS and commercial reference kits. The dual-T ICAS demonstrated good analytical sensitivity, procedural simplicity, and rapid detection capability. This ICAS can be used for early diagnosis and differentiation of DENV and CHIKV infections in medical institutions and individuals.