Key Laboratory of Quality and Safety Control of Citrus Fruits, Ministry of Agriculture and Rural Affairs, Southwest University, Chongqing, 400712, P. R. China.
National Citrus Engineering Research Center, Citrus Research Institute, Southwest University, Chongqing, 400712, P. R. China.
Anal Methods. 2024 Aug 15;16(32):5564-5570. doi: 10.1039/d4ay01150c.
The CRISPR/Cas12a system is a powerful signal amplification tool that has been widely used in nucleic acid detection. It has also been applied to the assay of non-nucleic acid targets, mainly relying on strategies for converting target determination into nucleic acid detection. Herein, we describe a CRISPR/Cas12a-based fluorescence method for sensitive detection of the total antioxidant capacity (TAC) by utilizing a strategy of converting TAC determination into Mn detection. Specifically, the reduction of MnO nanosheets by antioxidants produces plenty of Mn, which accelerates the -cleavage activity of CRISPR/Cas12a. Thus, a fluorescence enhanced detection method for TAC was established, with a detection limit as low as 0.04 mg L for a typical antioxidant, ascorbic acid. More importantly, this method has been proven to successfully analyze TAC in beverages. The excellent analytical performance of this method demonstrates the great potential of the CRISPR/Cas12a system in simple and sensitive TAC analysis.
CRISPR/Cas12a 系统是一种强大的信号放大工具，已被广泛应用于核酸检测。它也已被应用于非核酸靶标的测定，主要依赖于将靶标测定转化为核酸检测的策略。在此，我们描述了一种基于 CRISPR/Cas12a 的荧光法，通过将 TAC 测定转化为 Mn 检测来灵敏地检测总抗氧化能力（TAC）。具体而言，抗氧化剂将 MnO 纳米片还原会产生大量 Mn，从而加速 CRISPR/Cas12a 的切割活性。因此，建立了一种用于 TAC 的荧光增强检测方法，对于典型的抗氧化剂抗坏血酸，检测限低至 0.04mg L。更重要的是，该方法已被证明可成功分析饮料中的 TAC。该方法具有出色的分析性能，证明了 CRISPR/Cas12a 系统在简单灵敏的 TAC 分析中具有巨大的潜力。
