Abstract:To realize the visual toxicity screening of T-2 toxin, the fluorescent plasmid pcDNA 3.1-TRE-mCherry was constructed using the key AP-1 target response element TREand red fluorescent protein (mCherry) in the toxicity pathway of T-2 toxin and the sensing screening model of fluorescent HEK293 cells was established. To verify the applicability of the cell sensing model, the toxicity detection of T-2 toxin residue in the actual samples and HT-2 toxin which was the main metabolite of T-2 toxin was conducted. The results showed that the intracellular red fluorescence intensity reached the maximum value and tended to be stable when the T-2 toxin stimulated the model cells for 8 h. When the concentration of T-2 toxin was in the range of 1~25 ng/mL, it was linearly correlated with fluorescence intensity, and the linear equation was y=1.149 38x+64.72, R2，R2= 0.969. According to the dose curve, the EC50 of T-2 toxin was 16.27 ng/mL, and the detection limit was 0.691 ng/mL. The cell sensing model was used in the standard addition test. The average recovery of standard addition was 86.13%~126.20%, and the EC50 of HT-2 toxin was 27.65 ng/mL after toxicity screening.
