Nucleic acids have been engineered to participate in a wide variety of tasks. Among them, the enzyme-free amplification modes, enzyme-free DNA circuits (EFDCs), and hybridization chain reactions (HCRs) have been widely applied in a series of studies of bioanalysis. We demonstrated here an ultrasensitive hairpin probe-based circulation for continuous assemble of DNA probe. This strategy improved the analyte stability-dependent amplification efficiency of EFDC and signal enhancement without being limited by the analyte's initial concentration, and it was used to produce a novel microRNA (miRNA) trace analysis assay with ultrasensitive amplification properties. Through the detection of standard miRNA substances, 1 amol-level sensitivity and satisfactory specificity were achieved. Compared with EFDCs circulation was higher by 3 or 4 orders of magnitude. Furth demonstrated in the detection of miRNAs in tumor cells. The MCF-7 tumor cells were 10, 10, and 100 cells, respectively. In tissues. Thus, this ultrasensitive hairpin probe-based circulation field of tumor diagnosis. and HCRs, the sensitivity of ultrasensitive hairpin probe-based Furthermore, the excellent performance of this platform was also sensitivities for the detection of miRNAs in HepG2, A549 and addition, a high detection rate of 83% was achieved for tumor possesses the potential to be a technological innovation in the field of tumor diagnosis.