CRISPR/Cas12a-mediated Enzymatic recombinase amplification for rapid visual quantitative authentication of halal food

文献类型： 外文期刊

第一作者： Wang, Xiaohui

作者： Wang, Xiaohui;Jin, Wenyu;Yang, Yao;Ma, Huizi;Liu, Honghong;Lei, Jiawen;Zhang, Li;Yang, Yao;Wu, Yuhua

作者机构：

关键词： Adulteration; Porcine-derived ingredient; Rapid detection; CRISPR/Cas12a; Enzymatic recombinase amplification

期刊名称：ANALYTICA CHIMICA ACTA （ 影响因子：6.2； 五年影响因子：5.9 ）

ISSN： 0003-2670

年卷期： 2023 年 1255 卷

页码：

收录情况： SCI

摘要： Economically motivated adulteration (EMA) has become a concern in food safety. We propose a CRISPR/Cas12a Mediated Enzymatic Recombinase Amplification detection system (CAMERA) that integrates Enzymatic Recombinase Amplification (ERA) and Cas12a cleavage to detect halal food adulteration. We designed and screened crRNA targeting CLEC, a porcine-specific nuclear single-copy gene, and optimized the reagent concentrations and incubation times for the ERA and Cas12a cleavage steps. CAMERA was highly specific for pork ingredients detection. The DNA concentration and fluorescence signal intensity relationship was linear at DNA concentrations of 20-0.032 ng/mu L. CAMERA detected as few as two CLEC copies and quantified samples with porcine DNA content as low as 5% within 25 min. The system could be operated in a miniaturized working mode that requires no technical expertise or professional equipment, making CAMERA a valuable tool in resource-limited areas for the qualitative and quantitative detection of pork ingredients in halal food.

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