Background: The development of rapid and effective tools for the identification and quantitation of E. coli is of extreme importance in food analysis, environmental monitoring and clinical diagnostics.Methods: The biosensor developed in this work is based on electrochemistry reactions. Therefore, the layout of the sensor is patterned as a two-electrode configuration, working electrode (WE) and reference electrode (RE). The RE were prepared by mixing 1.20 g of graphite powder, which had been heated at 700oC in a muffle furnace for 15 s, with 800 mL of paraffin oil with a mortar and pestle. A WE was prepared in a similar fashion, except that the graphite powder was mixed with a desired weight of bacteriophage. Both RE and WE pastes were packed into a polyethylene tube (2.5 mm diameter), the tip of which had been cut off. Electrical contact to the paste was established via inserting a copper wire thorough flank.Results: Electrochemical experiments were carried out with an electrochemical interface LCR meter as a signal transducer and an electrochemical cell that contains the two-electrode system. The E. coli trapping on bacteriophage was reported by capacitance measurements. Conclusion: In this work, we have successfully fabricated an electrochemical biosensor with bacteriophage electrodes on a paste substrate. The proposed sensors have good characteristics such as; low detection limit, wide concentration, fast response time and good selectivity coefficient for Escherichia coli detection.