An amplifying DNA circuit coupled with Mg²⁺-dependent DNAzyme for bisphenol A detection in milk samples

As bisphenol A (BPA) is harmful to human health, it is of great significance to develop a new method for BPA detection. Herein, we designed a BPA biosensor by integrating an amplifying DNA circuit with Mg²⁺-dependent DNAzyme into the sensing system. The BPA-aptamer binding activated a DNA circuit for signal amplification based on toehold-mediated strand displacement. A catalytic Mg²⁺-dependent DNAzyme was formed through synergistically DNA hybridization, which can cleave the dual-labeled substrate DNA into two segments. The separation of the fluorophore and quencher produces a high fluorescence response for BPA detection. This biosensor exhibited a superior sensitivity with a detection limit of 50 fM. The method is selective and robust, which can work even in milk samples with satisfactory accuracy. The biosensor analytical results were also verified by liquid chromatography coupled with mass spectrometry (LC-MS) and no obvious difference existed between the two methods.