Rutin, as a biological flavonoid glycoside, has very important medicinal value. The accurate and rapid detection of rutin is of great significance. Herein, an ultrasensitive electrochemical rutin sensor based on β-cyclodextrin metal–organic framework/reduced graphene oxide (β-CD-Ni-MOF-74/rGO) was constructed. The obtained β-CD-Ni-MOF-74 was characterized by X-ray diffraction spectroscopy (XRD), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), and nitrogen adsorption and desorption. The β-CD-Ni-MOF-74/rGO presented good electrochemical properties benefiting from the large specific surface area and good adsorption enrichment effect of β-CD-Ni-MOF-74 and the good conductivity of rGO. Under optimal conditions for the detection of rutin, the β-CD-Ni-MOF-74/rGO/GCE showed a wider linear range (0.06–1.0 μM) and lower detection limit (LOD, 0.68 nM, (S/N = 3)). Furthermore, the sensor shows good accuracy and stability for the detection of rutin in actual samples.