We have developed a quadratic amplification strategy based on polymerase-aided strand-displacement polymerization (1-2) and exonuclease-assisted template recycling, which achieves rapid, isothermal, and highly sensitive detection of miRNAs extracted from cancer cell lines and clinical samples. Aside from sensitivity, our assay requires only one step to realize quadratic amplification for ultra-high sensitive detection of miRNAs, there is no need for multiple self-assembly steps required in nanoparticle-based amplification assay (3-4), complicated operation required in rolling circle amplification method (5-6). The proposed approach would be a promising tool for miRNAs research. For example, the early diagnosis of disease could be realized via quantitative studies of miRNA.

Lab in a tube: Ultra-sensitive detection of microRNAs in single cell and breast cancer patients using quadratic isothermal amplification

245th National Meeting (2013)

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