期刊
FOOD CHEMISTRY
卷 387, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.foodchem.2022.132919
关键词
Organophosphorus pesticides; Acetylcholinesterase; MnO2 nanosheets; DNAzyme; CRISPR-Cas12a
资金
- National Key Research and Development Program of China [2019YFC1605604]
- China Agriculture Research System [CARS-26]
In this study, a sensitive fluorescent assay for detection of organophosphorus pesticides (OPs) based on activating the CRISPR-Cas12a system is proposed. By utilizing the catalytic properties of acetylcholinesterase (AChE) and DNAzyme, the fluorescence signal is amplified for OPs detection. The method shows low detection limits and holds great potential for application in agricultural products.
Herein, we propose a sensitive fluorescent assay for organophosphorus pesticides (OPs) detection based on a novel strategy of activating the CRISPR-Cas12a system. Specifically, acetylcholinesterase (AChE) hydrolyzes acetylthiocholine into thiocholine (TCh). Subsequently, TCh induces the degradation of MnO2 nanosheets and generates sufficient Mn2+ ions to activate the Mn2+-dependent DNAzyme. Then, as the catalytic product of activated DNAzyme, the short DNA strand activates the CRISPR-Cas12a system to cleave the fluorophorequencher-labeled DNA reporter (FQ) probe effectively; thus, increasing the fluorescence intensity (FI) in the solution. However, in the presence of OPs, the activity of AChE is suppressed, resulting in a decrease in FI. Under optimized conditions, the limits of detection for paraoxon, dichlorvos, and demeton were 270, 406, and 218 pg/mL, respectively. Benefiting from the outstanding MnO2 nanosheets properties and three rounds of enzymatic signal amplification, the proposed fluorescence assay holds great potential for the detection of OPs in agricultural products.
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