A highly sensitive and accurate dual-channel fluorescent immunochromatographic assay for simultaneous quantitative detection of influenza A virus and adenovirus antigens

Sensitivity and accuracy are the key performance metrics for evaluating immunochromatographic assay (ICA). In this study, we have developed a quantum dot nanobeads-based dual-channel fluorescent ICA (QBs-DICA) designed for the sensitive and accurate detection of both influenza A virus (IAV) and human adenovirus (ADV) antigens. Following systematic assay optimization, we achieved a remarkable improvement in sensitivity, with gains of up to two orders of magnitude at the best. To enhance signal accuracy for IAV and ADV testing, we implemented a temperature-controlled dual-channel detector during sample incubation and the chromatographic process, resulting in an approximate increase of 34.2% and 36.7%, respectively. The Limit of Detection (LOD) for IAV and ADV testing was robustly established at 165 copies/mL and 805 copies/mL, respectively. Furthermore, in the case of IAV-spiked and ADV-spiked samples, we achieved consistently high recoveries ranging from 94.5% to 104.8% and 94.7% to 106.4%, with impressively low coefficients of variation (CV) of below 7.16% and 6.25%, respectively. Our established QBs-DICA system has exhibited a remarkable enhancement in both sensitivity and accuracy, making it exceptionally well-suited for supporting the rapid advancement of clinical diagnosis and treatment strategies.