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<p>Conventional detection methods for <i>Listeria monocytogenes</i> suffer from limitations including lengthy procedures, equipment dependency, and operational complexity, making them unsuitable for rapid on-site detection. Therefore, this study developed an LM-RPA-Cas12a-LFA detection method. Target bacteria were enriched using aptamer magnetic beads, followed by RPA amplification of a 196 bp fragment of the <i>hly</i> gene to activate the Cas12a system, with results displayed on lateral flow strips. The method including bacterial enrichment, DNA extraction, LM-RPA-Cas12a-LFA procedure required 2 h with detection limit of 1 × 10<sup>−10</sup> ng/µL and 1.35 CFU/mL in complex food matrices, demonstrating excellent reproducibility and stability. Detection of 16 real food samples showed complete concordance with qPCR validation, accurately identifying 4 positive and 12 negative samples. This method provides a reliable, rapid, and sensitive tool for on-site qualitative detection of <i>Listeria monocytogenes</i>.</p>

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Listeria monocytogenes detection assay via aptamer-functionalized magnetic bead enrichment coupled with RPA-CRISPR/Cas12a lateral flow strips

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年2月2日

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  • Han Wang,
  • Haosong Li,
  • Dexin Zeng,
  • Yansong Li,
  • Xiaofeng Yu,
  • Chunyi Shangguan,
  • Xilin Liu,
  • Xi Yang,
  • Honglin Ren,
  • Pan Hu,
  • Qiang Lu,
  • Shiying Lu

摘要

Conventional detection methods for Listeria monocytogenes suffer from limitations including lengthy procedures, equipment dependency, and operational complexity, making them unsuitable for rapid on-site detection. Therefore, this study developed an LM-RPA-Cas12a-LFA detection method. Target bacteria were enriched using aptamer magnetic beads, followed by RPA amplification of a 196 bp fragment of the hly gene to activate the Cas12a system, with results displayed on lateral flow strips. The method including bacterial enrichment, DNA extraction, LM-RPA-Cas12a-LFA procedure required 2 h with detection limit of 1 × 10−10 ng/µL and 1.35 CFU/mL in complex food matrices, demonstrating excellent reproducibility and stability. Detection of 16 real food samples showed complete concordance with qPCR validation, accurately identifying 4 positive and 12 negative samples. This method provides a reliable, rapid, and sensitive tool for on-site qualitative detection of Listeria monocytogenes.