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<p>The realization of sensitive detection of ofloxacin (OFL) residues is a key measure to prevent environmental risks caused by improper use of antibiotics, ensure food safety and promote public health security. Utilizing split crRNA with CRISPR/Cas12a (SCas12a)-powered cascade strand displacement amplification (C-SDA) and using phosphorothioate-modified G-quadruplex/triplex (psG43) concatemer fluorescence probe, a strategy named SCCA was devised for OFL detection. Target OFL triggers the SCas12a-driven C-SDA, and then activates SCas12a’s <i>trans</i>-cleavage activity. The activated SCas12a system cleaves reporter probe and then releases the psG43 sequence, which then combines with thioflavin T (ThT) to emit a fluorescence signal. The SCCA strategy not only avoids the design of PAM required by traditional C-SDA, but also can specifically detect OFL as low as 0.1&#xa0;pM. The anti-interference advantage of psG43 enables the utilization of the SCCA strategy for OFL detection in milk samples.</p>

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A Sensitive Aptasensor for Ofloxacin Detection Based on Split crRNA with CRISPR/Cas12a-Powered Cascade Strand Displacement and Phosphorothioate-Modified G-quadruplex/triplex Concatemer Fluorescence Probe

  • 期刊论文

  • 发表日期：

    2026年8月3日

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  • Hao Su,
  • Jiaye Wei,
  • Zhixiang Zhang,
  • Yu Ling,
  • Jiaxuan Chen,
  • Jiali Chang,
  • Kai Shi,
  • Tao Jiang

摘要

The realization of sensitive detection of ofloxacin (OFL) residues is a key measure to prevent environmental risks caused by improper use of antibiotics, ensure food safety and promote public health security. Utilizing split crRNA with CRISPR/Cas12a (SCas12a)-powered cascade strand displacement amplification (C-SDA) and using phosphorothioate-modified G-quadruplex/triplex (psG43) concatemer fluorescence probe, a strategy named SCCA was devised for OFL detection. Target OFL triggers the SCas12a-driven C-SDA, and then activates SCas12a’s trans-cleavage activity. The activated SCas12a system cleaves reporter probe and then releases the psG43 sequence, which then combines with thioflavin T (ThT) to emit a fluorescence signal. The SCCA strategy not only avoids the design of PAM required by traditional C-SDA, but also can specifically detect OFL as low as 0.1 pM. The anti-interference advantage of psG43 enables the utilization of the SCCA strategy for OFL detection in milk samples.