In this work, the fabrication and performances of a glucose biosensor based on the electrodeposited gold nanoparticle-polyvinylpyrrolidone-polyaniline (AuNP-PVP-PANI) nanocomposites have been demonstrated. The nanocomposites were electrodeposited on a glassy carbon electrode (GCE) in a homogeneous three-component solution consisting of aniline, PVP and AuNPs. Glucose oxidase (GOx) was subsequently immobilized on the as-prepared AuNP-PVP-PANI nanocomposites using Nafion as the preventing layer. The electrochemical and electrocatalytic properties of the Nafion/GOx/AuNP-PVP-PANI modified GCE were researched. It exhibited good electrocatalytic performances with a large linear range from 0.05 mM to 2.25 mM, a low detection limit of 1.0 x 10(-5) M (S/N = 3), and a high sensitivity of 9.62 mu A mM(-1) cm(-2). This sensor with excellent stability and reproducibility was also allowed for the detection of glucose in human serum samples with satisfactory results. (C) 2015 Elsevier B.V. All rights reserved.