Hairpin/DNA Ring Ternary Complex Initiated Rolling Circle Amplification for an Elevated Accuracy and Its Application...

Rolling circle amplification (RCA) is an attractive isothermal nucleic acid amplification approach and has been widely applied in constructing a variety of biosensors. However, the inevitable drawbacks of lacking enough selectivity greatly hindered further applications of RCA-based approaches. Here, we develop a novel RCA-based approach by integrating the specific target recognition capability of the hairpin/DNA ring ternary complex and multiple signal amplification and successfully applied it for let-7a detection. In this method, let-7a specifically unfolds the hairpin probe (Hp probe) in the ternary complex to induce target recycle and RCA- and DNAzyme-based signal generation. Based on this, the established approach exhibits a high selectivity to let-7a, and the response of the approach to one base pair mismatched sequences was 24.9%, indicating a significantly improved specificity. Meanwhile, the limit of detection is as low as 342 aM, which can meet the high requirement for a trace amount of miRNA detection. In all, we believe that the established approach can offer a new avenue for miRNA detection and post-tumor care.