Abstract
Trace antibiotic residues in animal foodstuff may lead to resistance and hazard human health, so simple and rapid detection methods have aroused multiple attention. Here, a novel bioluminescence pseudo-ELISA for the trace detection of multiple tetracyclines (TCs) in milk was designed and prepared based on a new fusion protein NLuc-TetR. The N-terminal of the bio-recognition motif TetR was linked to the C-terminal of the reporting motif nano-luciferase (NLuc) through SOE-PCR. Biolumescence signal showed that NLuc-TetR was specificity to nine TCs. And the acting pattern of substrate, NLuc-TetR, and TCs ternary complex was analyzed by molecular docking. With the optimal parameters, the half inhibition concentrations (IC50) of the developed bioluminescence pseudo-ELISA for nine TCs ranged from 0.39 to 1.44 ng mL−1, and the detection limits (LODs) ranged from 2 to 11 pg mL−1. The results demonstrated that the bioluminescence method based on NLuc-TetR is a promising method for routine screening measures for the residues of TCs in milk.
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