A new class of functionalized graphene oxide (FGO) nanosheet based amperometric glucose biosensor platform has been fabricated. FGO nanosheet comprises of chemically bound metalloid polymer hybrid (MPH) nanoparticles (average size of 12.5 +/- 2 nm) on the surface of a graphene oxide (GO) nanosheet. Spectroscopic characterization indicated that MPHs are well distributed, with a strong binding affinity between the GO nanosheets. The synergistic features of the metalloid polymer and the GO resulted in a unique three-dimensional nano-architecture on a gold-printed circuit board electrode (Au-PCB). The electrocatalytic response against a glucose sample is predominant, with a characteristic response time of 7 s, correlation co-efficient of 0.9981 and a wide linear range of up to 55.5 mM. The stability of the nanoarchiteciure modified on the electrode substrate is suitably durable for long-term application. The practical applicability of the fabricated electrode system was evaluated using a hyperglycemic clinical samples, and was compared with a commercial glucose biosensor. The obtained amperometric results were in good agreement with those of the commercial biosensor, and are promising for further clinical applications. (C) 2013 Elsevier Ltd. All rights reserved.