MicroRNAs (miRNAs), considered as therapeutic targets and biomarkers, play important roles in biological processes. Herein, an enzyme-free surface plasmon resonance imaging (SPRi) biosensing method has been developed for miRNA detection based on catalytic hairpin assembly and spherical nucleic acid. The hairpin H1 tethered on the surface of the sensor chip is unfolded by miRNA, and then the hybridized miRNA is released through the displacement of the hairpin H2 for the successive hybridization and assembly process. The emerging DNA fragments on the sensor chip surface after hairpins assembly are further used to hybridize with spherical nucleic acid, inducing a remarkably amplified SPR signal. This biosensing method is highly sensitive to miRNA with a detection limit of 53.7 fM and a linear range of 4 orders of magnitude. Moreover, the biosensor demonstrates good specificity and has the ability to distinguish members of homologous miRNA family even with single base differences. Thus, the SPRi biosensing method may hold a great promise for further application in early clinical diagnosis.
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