▎ 摘　　要

NOVELTY - Protein detecting film comprising nucleic acid aptamer, and substrate that specifically recognizes the protein, is new. The substrate comprises graphene film. The nucleic acid aptamer is attached to the surface of graphene film. USE - Used as protein detecting film. ADVANTAGE - The detecting film: uses single-layer graphene as response substrate of Raman spectroscopy, and selects Raman characteristic G peak associated with graphene phonon vibration mode as response signal; sensitively indicates the electronic structure and interface properties of single-layer graphene; avoids the complex marker of traditional Raman detection technology in the preparation process; and simplifies detection and operation procedures. DETAILED DESCRIPTION - INDEPENDENT CLAIMS are also included for: (1) preparing detecting film comprising, dropping phosphate buffer solution containing nucleic acid aptamer on the surface of substrate, drying the surface of substrate, washing the surface with water, and drying to obtain the protein detecting film; and (2) nucleic acid aptamer-graphene Raman frequency shift-based protein detection method, comprising (a) obtaining the protein detecting film, (b) detecting Raman spectrum of protein detecting film, and recording a G peak of graphene of protein detecting film, (c) adding solution to be tested to the surface of protein detecting film to which nucleic acid aptamer is attached, and culturing to prepare sample to be tested, (d) detecting Raman spectrum of sample to be tested, and recording a G peak of graphene of the sample to be tested, and (e) comparing the G peak position of graphene obtained in the step (b) with G peak position of graphene obtained in the step (d), judging based on the comparison result whether the solution to be tested contains the target protein corresponding to protein detecting film, and/or detecting the content of target protein.