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bioRxiv · 10.1101/2025.05.02.651981

CRISPR-On-Beads: A Simple and Sensitive Approach for Bacterial DNA Detection

Ye, J.·Liu, L.·Peng, R.·Xu, F.·Men, Y.·Du, K.
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Abstract

CRISPR/Cas12a, recognized for its high efficiency and specificity in nucleic acid recognition, has found extensive applications in infectious disease sensing. Nonetheless, a significant challenge persists in devising a simple method for the rapid, convenient, and accurate visualization of detection results. In this study, we demonstrate an innovative and simple CRISPR-on-beads assay for visual detection of methicillin-resistant Staphylococcus aureus (MRSA). This approach combines isothermal target amplification, CRISPR/LbCas12a-mediated cleavage, and magnetic bead-based probe binding for signal amplification, enabling rapid and convenient identification of bacterial DNA. Our results show that this detection method exhibits both high sensitivity and specificity, with a detection limit as low as 30 CFU/L. This newly developed biosensing approach can easily be integrated with a portable fluorescence microscope for medical diagnostics in resource-limited settings.

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Ye, J., Liu, L., Peng, R., Xu, F., Men, Y., Du, K.. 2025-05-02. CRISPR-On-Beads: A Simple and Sensitive Approach for Bacterial DNA Detection. https://doi.org/10.1101/2025.05.02.651981

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