"Turn-on" aptamer-immune lateral flow assays for the detection of small molecule targets based on CHA-assisted and CRISPR/Cas12a mediated signal transduction and amplification

文献类型： 外文期刊

第一作者： Dong, Haowei

作者： Dong, Haowei;Zhang, Pengwei;Lv, Liwen;Yu, Chunlei;Jia, Peng;Zhao, Jicheng;Du, Fangling;Guo, Yemin;Sun, Xia;Qin, Yifei;Du, Fangling

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关键词： Aptamer; Lateral flow assays; Small molecule targets; Turn-on; CRISPR/Cas12a

期刊名称：BIOSENSORS & BIOELECTRONICS （ 影响因子：10.5； 五年影响因子：10.1 ）

ISSN： 0956-5663

年卷期： 2025 年 285 卷

页码：

收录情况： SCI

摘要： Lateral flow assays (LFAs) have emerged as crucial tools for on-site food safety detection due to their simple operation and intuitive detection results. Nevertheless, LFAs for small molecule targets such as pesticides often present a "Turn-off" signal output, which leads to their low sensitivity and the risk of false positives. In this study, a CRISPR/Cas12a system-mediated strategy was employed to convert aptamer signals into the signals of immune LFAs, achieving a "Turn-on" signal output for highly sensitive detection of small molecule targets. The binding of aptamers to targets released the trigger sequence to initiate the catalytic hairpin assembly (CHA) reaction, generating double-stranded DNA, which subsequently activated the CRISPR/Cas12a system to cleave the FAMlabeled Reporter. Eventually, the "Turn-on" visual output of the signal was realized through an anti-6-FAM immune LFAs. The experiment optimized the sample pool preparation, CHA reaction conditions, CRISPR/ Cas12a activation parameters, and the assembly process of the LFAs. The limit of detection for procymidone was as low as 0.015 ng/mL, which was 52.67 times more sensitive than those of conventional aptamer-based LFAs without signal amplification strategies. This method exhibits high specificity for procymidone and a recovery rate ranging from 94.00% to 104.20% in vegetable samples, demonstrating excellent stability and practicability.

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