Abstract
Aflatoxin B1 (AFB1) is one of the most harmful mycotoxins. Convenient and reliable determination of trace AFB1 is highly desired. Herein, dual hemin labeled DNA hairpin was designed as a chemiluminescence (CL) switch and combined with aptamer of AFB1 and its partial complementary strand for homogenous assay of AFB1. Due to the judicious design of three DNA sequences and the help of nicking enzyme, the AFB1 recognition, signal transduction and recycling amplification can be implemented during one-step reaction. The whole protocol only involved the simple mixing of samples with homogeneous solution, avoiding the preparation of sophisticated solid-phase sensor and eliminating the multi-step washing and resulting errors, which provided a unique tool for on-site rapid testing. Importantly, when the operation process was simplified, the detection sensitivity can be enhanced facilely due to the enzyme recycling. Under optimal conditions, the intelligent CL system for sensing AFB1 exhibited a wide dynamic range (1.0 × 10-2 ∼ 100 ng mL−1) with a low detection limit (down to 1.5 pg mL−1), suitable for detecting trace AFB1 in complex matrix. The methodology was validated in coix seed and decoction pieces. The superiority including free of washing and separation, high sensitivity, acceptable accuracy, and greenness, showed good applicability of the proposed CL system in safety evaluation of food and medicine.
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