In order to realize the highly specific and sensitive detection for miRNA, a multiple miRNA quantitative detection protocol was designed by combined the fluorescence characteristics of quantum dots with the DNA shearing properties of double-strand specific nuclease. First, the two ends of the capture DNA are separately linked with the quantum dots and the ferroferric oxide magnetic nanoparticles to form a capture probe, and then the capture DNA and the detected miRNA will form a heteroduplex DNA-miRNA hybrid structure as they are complementary gene sequences. And then, the DNA is specifically sheared by double-strand specific nuclease to separate the quantum dots and the detected miRNA from the capture probe, and the isolated miRNA and the unpaired capture probe start a new round of hybridization and specific shear of the double-strand specific nuclease. Through the above cyclic process, quantum dots are continuously released from the capture probe, resulting in the continuous enhancement of the detected fluorescent signal, thereby achieving highly sensitive detection of tumor marker miRNA. The experimental results show that the specific quantitative detection of miRNA-141 and miRNA-1228 is successfully achieved by double-strand specific nuclease-assisted circular amplification of quantum dot fluorescence signals in the concentration range of 1 fmol/L to 100 pmol/L, and the detection limits reached 0.69 fmol/L and 0.21 fmol/L, respectively. In addition, compared with the real-time fluorescence quantitative polynucleotide chain reaction method, the proposed protocol obtains consistent detection results with a higher sensitivity.