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In this work, a sensor for quantitative detection of uric acid (UA) is successfully prepared by electrodepositing Nickel–Copper (Ni–Cu) alloy nanoparticles on poly (3,4-ethyldioxythiophene) modified nitrogen-containing grapheme (Ni–Cu/PEDOT/NGE). The anchoring of PEDOT and Cu–Ni alloy nanoparticles on NGE not only prevents the agglomeration of NGE, but also improves the catalytic activity of the composites. Ni–Cu/PEDOT/NGE shows high electrochemical performance to UA and the oxidation of UA on its interface was an action with two protons and two electrons. Under the optimized condition, the response current of the sensor is linear with UA concentration within the range of 0.1–10 μ M and 10–50 μ M. A low detection limit of 0.059 μ M at S/ N = 3 is obtained. Additionally, the fabricated electrochemical sensor with good sensitivity and selectivity to UA may be promising for practical clinical testing. Poly(3,4-ethylenedioxythiophene) and nickel–copper alloy nanoparticles are modified on nitrogen-doped graphene via electropolymerization and electrodeposition (Ni–Cu/PEDOT/NGE). The oxidation of uric acid on the interface of Ni–Cu/PEDOT/NGE is an action with two protons and two electrons. [ABSTRACT FROM AUTHOR]