A novel and simple synchronous electrochemical synthesis of poly(xanthurenic acid, Xa), electrochemically reduced graphene oxide nanocomposite (PXa-ERGNO), via cyclic voltammetry (CV) was reported, where graphene oxide (GNO) and Xa monomer were adopted as precursors. The resulting PXa-ERGNO nanocomposite was characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, CV and electrochemical impedance spectroscopy (EIS). The π-π interactions between the conjugated GNO layers and aromatic ring of Xa-enhanced the electropolymerization efficiency accompanied with an increased electrochemical response of PXa. The rich carboxyl groups of PXa-ERGNO film were applied to stably immobilize the probe DNA with amino groups at 5' end via covalent bonding. The captured probe could sensitively and selectively recognize its target DNA via EIS. The dynamic detection range was from 1.0 × 10(-14) mol/L to 1.0 × 10(-8) mol/L with a detection limit of 4.2 × 10(-15) mol/L due to the synergistic effect of integrated PXa-ERGNO nanocomposite. This graphene-based electrochemical platform showed intrinsic advantage, such as simplicity, good stability, and high sensitivity, which could serve as an ideal platform for the biosensing field.