Development Of Test Strip For Quantitative Detection Of Mycotoxins Based On Fluorescence Immunochromatographic Assay

In this study,fluorescent microsphere materials with characteristic emission peak at525 nm were used as signal markers to couple antibodies of ZEN,AFB₁,FB₁,and OTA to prepare fluorescent probes.Chicken egg albumin（OVA）was conjugated with four toxin haptens by activated ester method to prepare the corresponding coating antigens.Based on the specific recognition of antigen and antibody,fluorescence immunochromatographic assay were established,which realized the rapid quantitative detection of four mycotoxins in grains.For the establishment of fluorescence immunochromatographic assay for ZEN,the antibody dosage during coupling of fluorescent microspheres with antibody,the type of NC membrane,the kind of loading buffer,the dilution ratio of goat anti mouse Ig G₁and coating antigen,and the amount of FMs-Ab were optimized.Under the optimal detection conditions,the detection linear range was from 0.14 to 1.93μg·L^-1and the detection limit was 0.072μg·L^-1.For the establishment of fluorescence immunochromatographic assay for AFB₁,the antibody dosage during coupling of fluorescent microspheres with antibody,the type of NC membrane,the kind of loading buffer,the dilution ratio of goat anti mouse Ig G₁and coating antigen,and the amount of FMs-Ab were optimized.Under the optimal detection conditions,the detection linear range was from 0.2 to 3.69μg·L^-1and the detection limit was 0.093μg·L^-1.For the establishment of fluorescence immunochromatographic assay for FB₁,the antibody dosage during coupling of fluorescent microspheres with antibody,the type of NC membrane,the kind of loading buffer,the dilution ratio of goat anti mouse Ig A and coating antigen,and the amount of FMs-Ab were optimized.Under the optimal detection conditions,the detection linear range was from 0.66 to 11.66μg·L^-1and the detection limit was 0.32μg·L^-1.For the establishment of fluorescence immunochromatographic assay for OTA,the antibody dosage during coupling of fluorescent microspheres with antibody,the type of NC membrane,the kind of loading buffer,the dilution ratio of goat anti mouse Ig G₂and coating antigen,and the amount of FMs-Ab were optimized.Under the optimal detection conditions,the detection linear range was from 0.47 to 15.89μg·L^-1and the detection limit was 0.19μg·L^-1.The fluorescence immunochromatographic assay has good specificity,and UPLC-MS/MS was used to verify the accuracy of the detection method established in this study,which achieved good consistency.This method is suitable for the rapid quantitative detection of mycotoxins in grains.