Chaochao Chen,Liang Luo,Jianzhong Shen,Zhanhui Wang,Yantong Pan
The enzyme-linked immunosorbent assay (ELISA) stands as one of the most frequently employed rapid detection techniques for both chemical and biological contaminants. Horseradish peroxidase (HRP) and hydrogen peroxide (H2O2), serving as signal generators, play indispensable roles throughout the entire ELISA process. However, HRP and H2O2 exhibit high sensitivity to elevated temperatures, hindering the broader utilization and transportation of ELISA. Hereby, being inspired by the cascade catalysis of glucose by glucose oxidase (GOx) and HRP as well as the advantageous properties of metal–organic frameworks (MOFs), we developed a cascade catalysis-based competitive fluorescent ELISA (CCF-ELISA) for mycotoxin detection in maize using zearalenone (ZEN) as a model. The results revealed the remarkable protective effects of MOFs on enzymatic activities. Based on upon characteristics, the built CCF-ELISA enabled the detection of ZEN with a half-maximal inhibitory concentration of 0.54 ng/mL and allowed a detection range from 0.19 to 1.51 ng/mL. When applied to naturally contaminated maize samples, the outcomes obtained from the CCF-ELISA showed a high level of comparability with high-performance liquid chromatography–tandem mass spectrometry, demonstrating the promising potential of the developed CCF-ELISA in the rapid detection for food security.
Dibutyl phthalate (DBP) is considered as one of the most widely used phthalate esters (PAEs), which has attracted worldwide concerns because of its potential threats to eco-environments and human health. Systematic investigations of DBP environmental occurrence contribute to the further risk assessment, which depends on effective and available analytical methods. In this study, an amplified ratiometric fluorescence ELISA was established for sensitive and high-throughput detection of DBP in the aquatic system based on a novel tetrahedral DNA nanostructure (TDN)-scaffolded-DNAzyme (Tetrazyme). Wherein, Tetrazyme was prepared by the precise folding of G-quadruplex sequence on three vertex angles of the TDN, together with hemin as the horseradish peroxidase (HRP)-mimicking enzyme. The rigid TDN avoided the local overcrowding effect to provide a reasonable spatial spacing on the interface for G-quadruplex sequence, increasing the collision chance between DNAzyme and substrates, improving the catalytic ability of DNAzyme effectively. Besides, streptavidin (SA) and biotin (bio) were used to anchor TDN and antibody, in which the specific binding of SA/bio could make more Tetrazyme conjugate on each signal element, resulting in the dual signal amplification. Meanwhile, the accuracy and precision were enhanced owing to the inherent built-in rectification to the environment from the dual output ratiometric fluorescence assay. Under the optimized conditions, the detection limit of this proposed method was 0.17 ng/mL (16 times lower than that of conventional ELISA using the same antibody) with a satisfactory accuracy (recoveries, 79.0%- 116.2%; CV, 2.1–6.5%). Overall, this platform provides a promising way for accurate, sensitive and rapid determination of DBP from environmental waters.
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