A highly sensitive immunofluorescence sensor based on bicolor upconversion and magnetic separation for simultaneous...

Mycotoxins cause significant harm to human health, so it is imperative to develop a highly sensitive and easy-to-operate method for the detection of mycotoxins. Herein, a fluorescence-based magnetic separation immunoassay for simultaneous detection of mycotoxins fumonisin B1 and zearalenone is established. The method employed high fluorescent upconversion-nanoparticles(UCNPs) conjugated with biotinylated antigens as upconversion fluoroscent probes. Magnetic nanoparticles(MNPs) immobilized with monoclonal antibodies are used as immune-capture probes. Highly sensitive detection of FB1 and ZEN was achieved based on the luminescence properties of UCNPs and the separation effects of MNPs. The results showed a robust linear correlation between the enhanced fluorescence emission intensity and the logarithmic concentrations of FB1 and ZEN under the optimal conditions (R²(FB1) = 0.9965, R²(ZEN) = 0.9976), and the linear ranges were 0.05-5 ng mL^-1. Furthermore, the limits of detection (LOD) were 0.016 ng mL^-1 for FB1 and 0.012 ng mL^-1 for ZEN. The standard addition method was used to determine the content of FB1 and ZEN in the samples to evaluate the accuracy of the process. The average recoveries were 89.48% to 113.69% and 85.97% to 113.82%, respectively. Compared with the other five mycotoxins, this method had high selectivity. It is expected that the multi-component simultaneous detection can be further realized by using the multicolor labeling characteristics of UCNPs.