A novel composite structure of reduced graphene oxide (rGO)-Ag nanoparticles (Ag NPs) nanocomposite, which was integrated on the indium tin oxide (ITO) glass by a facile and rapid two-step chronoamperometry electrodeposition route, was proposed and developed in this paper. SERS-activity of the rGO/Ag NPs nanocomposite was mainly affected by the structure and size of the fabricated rGO/Ag NPs nanocomposite. In the experiments, the operational conditions of electrodeposition process were studied in details. The electrodeposited time was the important controllable factor, which decided the particle size and surface coverage of the deposited Ag NPs on ITO glass. Under the optimized conditions, the detection limit for rhodamine6G (R6G) was as low as 10(-11) M and the Raman enhancement factor was as large as 5.9 x 10(8), which was 24 times higher than that for the ITO-Ag NPs substrate. Apart from this higher enhancement effect, it was also illustrated that extremely good uniformity and reproducibility with low standard deviation could be obtained by the prepared ITO-rGO/Ag NPs nanocomposite for SRES detection. (C) 2015 Elsevier B.V. All rights reserved.