In this article, we propose a novel microfluidic microstrip electromagnetic band gap (EBG) sensor realized using cost-effective 3D printing technology. Microstrip sensor allows monitoring of the fluid properties flowing in the microchannel embedded between the microstrip line and ground plane. The sensor\u2019s operating principle is based on the phase-shift method, which allows the characterization at a single operating frequency of 6 GHz. The defected electromagnetic band gap (EBG) structure is realized as a pattern in the microstrip ground plane to improve sensor sensitivity. The designed microfluidic channel is fabricated using a fused deposition modelling (FDM) 3D printing process without additional supporting layers, while the conductive layers are realized using sticky aluminium tape. The measurement results show that the change of permittivity of the fluid in the microfluidic channel from 1 to 80 results in the phase-shift difference of almost 90\u00b0. The potential application is demonstrated through the implementation of a proposed sensor for the detection of toluene concentration in toluene\u2013methanol mixture where various concentrations of toluene were analysed.