A low-cost and enzyme-free glucose paper sensor is presented as a promising alternative to glucose test strips. This paper-based glucose sensor is prepared with molecularly imprinted (MIP) polyaniline (PANI) electrode. The determination of glucose concentrations was studied by the impedance change of the paper sensor before and after the blood samples dispensing at a low frequency. A comparison of the linear and polynomial regression was applied to analyze the impedance ratio as a function of glucose concentrations. The proposed glucose paper sensor showed a limit of detection (LoD) of 1.135 mM. This novel and non-enzymatic paper sensor suggests a low-cost glucose test assay and can improve the quality of routine testing for diabetic patients.