CRISPR-on-beads: A simple and sensitive assay for detection of methicillin-resistant staphylococcus aureus

• A novel and simplified CRISPR-on-beads assay for the visual detection of methicillin-resistant Staphylococcus aureus (MRSA) is introduced. • Magnetic bead-based probe binding for enhanced signal amplification is developed, allowing for quick and convenient identification of bacterial DNA. • A detection limit as low as 30 CFU/μL is achieved, demonstrating excellent sensitivity and specificity. 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/μL. This newly developed biosensing approach can easily be integrated with a portable fluorescence microscope for medical diagnostics in resource-limited settings.