A novel piezoelelctric biosensor for the detection of Cholera Toxin (CT) has been developed, based on a lipid layer with GM1(Monosialoganglioside) and the subsequent binding of the specifically agglutinated network complex of GM1-incorporated liposomes in assay medium after the characteristic recognition of CT by the sensing interface. A lipid monolayer comprising a fixed DPPC: GM1 molar ratio was spontaneously formed on the gold surface by exposing it to an aqueous dispersion of GM1-incorporated phospholipid vesicles. The quartz crystal surfaces with GM1-containing supported lipid membranes can detect CT and probe the mass change on the sensing interface. The simple preparation of sensing interface with lipid monolayer makes the developed technique especially useful for rapid and renewable immunochemical determination. The frequency responses of the developed piezoelelctric biosensors are linearly correlated to CT concentration in the range of 0.25 – 1.0 colong mL-1 with a detection limit of 95ng mL-1. This could be practically used for the detection of CT.