The frequent detection of physiological glucose levels from human blood or sweat requires the development of low-cost electrodes with high sensitivity and selectivity. In this work, we prepared a series of Prussian blue (PB) modified carbon cloth (CC) electrodes with different cyanoferrate groups. We achieved a sensitivity as high as 145.43 μA mm−1cm−2 in a 0.1–6.5 mm concentration range with a response time below 2 s under physiological pH. The electrodes exhibited a superior selectivity of glucose in the presence of interfering agents, including sucrose, lactose, NaCl, ascorbic acid, and uric acid. The electrodes also showed outstanding long-term stability over 15 days. Furthermore, we performed comprehensive electrochemical and characterization studies to elucidate the role of the cyanoferrate group on the morphologic and electronic properties of non-enzymatic glucose sensors.