Abstract:

OBJECTIVE: To establish time-resolved immunofluorescence analysis (TRIFA) kit used to rapidly detect fumonisin B1 (FB1) in food. METHODS：Ascites was obtained after intraperitoneal injection of hybridoma cell lines against monoclonal antibody of FB1 into the mice，and the ascites was purified with protein A affinity chromatography so as to get a large amount of monoclonal antibody. Based on this antibody，the TRFIA kit was established and parameters including the detection limit，specificity，stability，recovery，repeatability and reproducibility were optimized. Nineteen corn samples and one blind sample of corn were detected with the established kit and the results were verified with the commercially available ELISA-kit. RESULTS：The detection limit of the established kit was 2 ng/mL，the linear range of detection was 2-512 ng/mL，the linear equation was Y=-0.644X+12.872 (R2=0.998)，and the 50% concentration of inhibition was 10.07 ng/mL. The rate of recovery from corn samples ranged from 78.32% to 116.76%. There was no interaction with deoxynivalenol，aflatoxins A and BSA. At room temperature，the kit could be kept for more than 315 days. CONCLUSION：The fast and sensitive time-resolved immunofluorescence analysis assay was established to detect FB1 in food.

Key words: fumonisin B1, monoclonal antibody, time-resolved immunofluorescence, TRFIA kit