A CRISPR/Cas12a-based fluorescence method for the amplified detection of total antioxidant capacity†

IF 2.7 3区化学 Q2 CHEMISTRY, ANALYTICAL

Analytical Methods Pub Date : 2024-07-29 DOI:10.1039/D4AY01150C

Qi Wu, Longyingzi Xie, Lanrui Ma, Xinqi Long, Lei Liu, Aihua Chen, Yongliang Cui, Yaohai Zhang and Yue He

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Abstract

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 Mn²⁺ detection. Specifically, the reduction of MnO₂ nanosheets by antioxidants produces plenty of Mn²⁺, which accelerates the trans-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⁻¹ 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.

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基于 CRISPR/Cas12a 的荧光方法，用于扩增检测总抗氧化能力。

CRISPR/Cas12a 系统是一种强大的信号放大工具，已广泛应用于核酸检测。它也被应用于非核酸靶标的检测，主要依赖于将靶标确定转化为核酸检测的策略。在此，我们介绍了一种基于CRISPR/Cas12a的荧光方法，利用将总抗氧化能力（TAC）测定转化为Mn2+检测的策略，灵敏地检测总抗氧化能力（TAC）。具体来说，抗氧化剂还原 MnO2 纳米片会产生大量 Mn2+，从而加速 CRISPR/Cas12a 的反式裂解活性。因此，我们建立了一种荧光增强的 TAC 检测方法，对典型的抗氧化剂抗坏血酸的检测限低至 0.04 mg L-1。更重要的是，该方法已被证明可成功分析饮料中的 TAC。该方法的卓越分析性能证明了 CRISPR/Cas12a 系统在简单灵敏的 TAC 分析中的巨大潜力。

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