Xiaoyan Li,Yang Qi,Changxu Zhao,Fei Wang,Yanlei Ding,Guangzheng Hou,Pengcheng Cao,Zhe Zhou,Qiqi Liu
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.
Zongwen Wang,Xiating Li,Feng Zhang,Yu Gao,Jintian Cheng,FengFu Fu
It is an urgent need to develop simple and high-throughput methods for simultaneously screening and detecting multiple or groups of sulfonamides (SAs) in animal-derived foods since various SAs were alternately used in animal husbandry to avoid generating drug resistance. We herein developed a novel HCl-reduced nicotinamide adenine dinucleotide I (NADH)-ascorbic acid (AA)-mediated gold nanobipyramids (AuNBPs) growth system, which can precisely regulate the growth rate of AuNBPs, to generate two colorful and stable AA-corresponding multicolor signal channels with different sensitivities. Based on the HCl-NADH-AA-mediated AuNBP growth system, we further developed a dual-channel multicolor immunoassay for simultaneously realizing rapid screening and detection of 5 SAs (sulfamethazine, sulfamethoxydiazine, sulfisomidine, sulfamerazine, and sulfamonomethoxine) by using a paper-based analytical device for sensitively and stably reading out the signal and a broad-specificity anti-SAs antibody as a bio-receptor. The developed immunoassay has more color changes, a wider linear range, excellent specificity and stability, and two multicolor signal channels (L-channel and H-channel) with different sensitivities. The H-channel exhibited 7–8 SAs-corresponding color changes and can be used to detect 5 target SAs with a visual detection limit of 0.1–0.5 ng/mL and a spectrometry detection limit of 0.05–0.16 ng/mL. The L-channel exhibited 7–9 SAs-corresponding color changes and can be used to detect 5 target SAs with a visual detection limit of 2.0–6.0 ng/mL and a spectrometry detection limit of 0.40–1.47 ng/mL. The developed immunoassay was successfully used to simultaneously screen and detect low-concentration and high-concentration of target SAs in milk and fish muscle samples with a recovery of 85–110% and an RSD (n = 5) < 8%. The visual detection limit of our immunoassay is much lower than the maximum residue limit of total SAs in edible tissue. All above features make our immunoassay a promising assay for simultaneously realizing the rapid screening and quantitative determination of multiple SA residues in food by bare eye observation. It must be mentioned that our immunoassay may be expended as a general method for the simultaneous visual screening and detection of other drugs using the corresponding antibody as a recognition probe.
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