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<p>The development of a detection technique with superior accuracy, high sensitivity, and visual applicability simultaneously for economically motivated adulteration (EMA) of fruit juice remains a formidable challenge. Herein, we developed a CRISPR/Cas12a-regulated fluorescence detection platform capable of converting trace level of target into detectable fluorescence signals. This strategy was inspired by the extraordinary <i>trans-</i>cleavage capacity of CRISPR/Cas12a and significant fluorescence response of the fluorescence probe [Ru(phen)<sub>2</sub>(dppz)]<sup>2+</sup>. After the target DNA had been identified, activated CRISPR/Cas12a cleaved the block ssDNA (bDNA), preventing the formation of short dsDNA and initiating a hybridization chain reaction (HCR). Consequently, large amounts of [Ru(phen)<sub>2</sub>(dppz)]<sup>2+</sup> intercalated into the resulting long dsDNA nanowires, enabling cascade signal amplification and a&#xa0;highly sensitive signal output. The output could be observed as a strong orange fluorescence only using a blue transilluminator. Then, we used the strategy to detect adulteration in blueberry juice, which displayed excellent specificity and satisfactory sensitivity with the limit of detection (LOD) as low as 19.5 fM. Our study demonstrated a novel method utilizing the fluorescence switch regulated by CRISPR/Cas12a, showing extremely high specificity, excellent sensitivity, and capability for visual detection of trace target substances in complex food matrices.</p> Graphical Abstract <p></p>

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CRISPR/Cas12a-Based Biosensor for Visual and Highly Sensitive Detection of Juice Adulteration via Structurally Defined Fluorescence Probe

  • 期刊论文

  • 发表日期：

    2026年6月15日

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  • Yue Yang,
  • Jianghua Liu,
  • Fulu Liu,
  • Jianing Chen,
  • Qianyu Li,
  • Bufan Xu,
  • Qinfeng Xu,
  • Guoliang Li

摘要

The development of a detection technique with superior accuracy, high sensitivity, and visual applicability simultaneously for economically motivated adulteration (EMA) of fruit juice remains a formidable challenge. Herein, we developed a CRISPR/Cas12a-regulated fluorescence detection platform capable of converting trace level of target into detectable fluorescence signals. This strategy was inspired by the extraordinary trans-cleavage capacity of CRISPR/Cas12a and significant fluorescence response of the fluorescence probe [Ru(phen)2(dppz)]2+. After the target DNA had been identified, activated CRISPR/Cas12a cleaved the block ssDNA (bDNA), preventing the formation of short dsDNA and initiating a hybridization chain reaction (HCR). Consequently, large amounts of [Ru(phen)2(dppz)]2+ intercalated into the resulting long dsDNA nanowires, enabling cascade signal amplification and a highly sensitive signal output. The output could be observed as a strong orange fluorescence only using a blue transilluminator. Then, we used the strategy to detect adulteration in blueberry juice, which displayed excellent specificity and satisfactory sensitivity with the limit of detection (LOD) as low as 19.5 fM. Our study demonstrated a novel method utilizing the fluorescence switch regulated by CRISPR/Cas12a, showing extremely high specificity, excellent sensitivity, and capability for visual detection of trace target substances in complex food matrices.

Graphical Abstract