The invention belongs to protein detection in the biomedical field, and in particular provides a DNA helicase detection method based on a graphene oxide chip. The DNA helicase detection method based on the graphene oxide chip comprises the following steps: (1), preparing a slide with graphene oxide dot matrixes; (2), connecting a double-chain DNA sequence with the slide with the graphene oxide dot matrixes; (3) detecting DNA helicase, to be specific, adding different quantities of DNA helicases at different action time, performing PBS washing to scan and analyze, and detecting according to FAM fluorescence intensity change. According to the DNA helicase detection method based on the graphene oxide chip, fluorescene on FAM marked by a single-stranded DNA can be quenched by utilizing a graphene oxide; after the DNA helicase is applied to a double-stranded DNA, a single-stranded DNA separates from the complementary single-stranded DNA fixed on the graphene oxide, and the distance between the FAM and the surface of the graphene oxide is changed, so that the change of FAM fluorescence intensity is caused; by combining a high-throughput microarray chip technique, the DNA helicase can be detected with low cost and high sensitivity.