In this work, based on a dual signal amplification strategy of enzyme-assisted signal amplification (EASA) and catalytic hairpin assembly (CHA), combined with the magnetic attraction effect, a capillary pump-driven surface-enhanced Raman scattering (SERS) microfluidic chip (LoC-SERS) platform was developed for the sensitive detection of colorectal cancer-associated (CRC) microRNA (miRNA). During the detection process, the miRNA first undergoes an EASA reaction with hairpin DNA1 (hpDNA1) under the action of endonuclease, which generates a large amount of DNA2 cyclically. After that, DNA2 triggers the CHA reaction to proceed, which leads to the ligation of the SERS nanoprobes and the capture nanoprobes (hpDNA2-hpDNA3 complexes). Finally, as the reactant solution flows through the collection zone, the end products are magnetically attracted by the micro-magnets, generating many "hot spots" and leading to a triple amplification of the SERS signal. By quantitative analysis, the platform achieved ultra-low detection limits of miR-122 (4.26 aM) and miR-192 (4.71 aM) within a linear range of 10 aM-10 pM. In addition, the platform's results for clinical samples are highly consistent with those measured by qRT-PCR methods. Overall, the proposed LoC-SERS platform is expected to be an important tool for the early screening of CRC.
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