In this study, a hybrid nanocomposite consisting of a conducting polymer and gold nanoparticles (AuNPs) is fabricated onto a screen-printed carbon electrode (SPCE). A thin layer of poly(3,4-ethylenedioxythiophene)(PEDOT) is coated electrochemically on a bare SPCE; then, the nano-sized AuNPs are embedded by electrochemical deposition. The resultant SPCE/PEDOT/AuNPs-modified electrode is characterized by electrochemical methods, field emission scanning electron microscopy (FE-SEM) and X-ray photoelectron spectroscopy (XPS). The SPCE/PEDOT/AuNPs-modified electrode possesses great catalytic activity for the oxidation of cysteine in various pH buffer solutions (pH 2.0-8.0). The selectivity of the method is demonstrated by the separation of the oxidation peaks at up to 240 mV for cysteine and glutathione in pH 6.0 buffer solutions. The effects of the oxidizable interferences are also investigated. Flow-injection amperometry is performed for 0.5-200 mu M of cysteine in pH 4.0 buffer solutions, and a linear calibration plot with a slope of 0.115 mu A/mu M is obtained. The detection limit (S/N = 3) is 0.05 mu M. Additionally, the proposed methods obtain satisfactory results in the detection of cysteine-containing medicine samples. (C) 2011 Elsevier Ltd. All rights reserved.