CRISPR/Cas12a assay for amol level microRNA by combining enzyme-free amplification and single particle analysis

CRISPR/Cas systems are increasingly utilized for sensitive miRNA detection through enzyme-based pre-amplification. To address challenges such as high costs, non-specific amplification, and interference from primer residues in pre-amplification strategies, herein a dual amplification CRISPR miRNA assay was developed by combining enzyme-free HCR with single-particle analysis. Attomolar detection limits, excellent selectivity, and practicability were achieved by applying this method.