A PSA (Prostate Specific Antigen) high-sensitivity detection method based on nanotechnology is a novel biological high-sensitivity detection method which combines the magnetic bead probe technology, Au-nanoparticle double-mark probe technology, enzyme-linked immunosorbent assay technology and the nucleic acid amplification technology together. According to the detection method, an antibody corresponding to a protein to be detected (such as PSA) is cross-linked with a magnetic bead, and after the protein to be detected in a sample is captured, an Au-nanoparticle dual-mark probe marked by another antibody and nucleic acid is added to the captured protein. After magnetic separation and purification are carried out, nucleic acid undergoes release amplification, a visual result is obtained by means of agarose gel electrophoresis, and the grey level value scanning is subsequently performed. Compared with the conventional ELISA (Enzyme-Linked Immunosorbent Assay), the detection sensitivity of the detection method disclosed by the invention is increased by six orders of magnitude, and the PSA high-sensitivity detection method based on nanotechnology is fairly significant for the early diagnosis of caners and the detection of trace important substances, nucleic acids and proteins.