Sensitive detection of microRNAs (miRNAs) that serve as a disease marker could advance the diagnosis and treatment of diseases. Many methods used for quantitative detection of miRNAs, such as PCR-based… Click to show full abstract
Sensitive detection of microRNAs (miRNAs) that serve as a disease marker could advance the diagnosis and treatment of diseases. Many methods used for quantitative detection of miRNAs, such as PCR-based approaches or the hybridization chain reaction, have presented challenges due to the complicated and time-consuming-procedures that are required. In this manuscript, a simple triggerable mutually amplified signal (TMAS) probe was designed and enriched within the center of a microfluidic chip and then used for one-step quantitative detection of microRNAs via surface enhanced Raman scattering (SERS) technology. First, many mutually amplified double strands are produced via an enzyme-free target-strand displacement recycling reaction initiated by the target miRNA, that result in the generation of an enhanced SERS signal. Second, microfluidic chips that utilize alternating current (AC) electrokinetic flow technology produce efficient mixing and rapid concentration to improve the DNA hybridization rate and further enhance the SERS signal intensity. This method enables the sensitive and rapid detection of miR-21 in human breast cancer cells within 30 min with a detection limit of 2.33 fM. Compared with traditional methods, this novel method overcomes the shortcomings resulting from complex operations, and has the advantages of high sensitivity, short assay time, and reduced sample usage.
               
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