Nanoporous Co3O4 nanowires (NCoNWs) were synthesized via a two-step process with hydrothermal growth of cobalt nanowires (NWs) precursors and subsequent calcination of corresponding precursors in air. The structures and morphologies of the resulting cobalt NWs precursors and NCoNWs were characterized by X-ray diffraction (XRD), scanning electron microscopic (SEM) and transmission electron microscopic (TEM). NCoNWs show high electrocatalytic activity toward glucose oxidation in alkaline medium and be used for non-enzymatic electrochemical detection of glucose. The sensor displays a high sensitivity of 300.8μAmM−1cm−2, a low detection limit of 5μM (S/N=3), and a wide linear dynamic range (5.0×10−6–5.7×10−4M). Furthermore, the sensor exhibits a high selectivity for glucose against the common interferences including ascorbic acid, uric acid and dopamine which co-exist with glucose in human serum samples.