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February 5, 2026npj Science of Food0 citationsOpen Access

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

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HWHan WangHLHaosong LiDZDexin Zeng

Key Points

  • The aim is to create a quick and efficient method for detecting Listeria monocytogenes in food samples.
  • Enrichment of target bacteria using aptamer-functionalized magnetic beads.
  • RPA amplification of a specific fragment of the hly gene.
  • Activation of the Cas12a system for detection.
  • Results displayed on lateral flow strips.
  • The method requires only 2 hours for completion.
  • Achieved a detection limit of 1 × 10^-10 ng/µL and 1.35 CFU/mL.
  • Complete concordance with qPCR validation in 16 real food samples.
  • Accurately identified 4 positive and 12 negative samples.

Abstract

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.

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Cite This Study

Wang et al. (2026) studied this question.

synapsesocial.com/papers/698433c8f1d9ada3c1fb13e7https://doi.org/10.1038/s41538-026-00728-4
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