This paper demonstrated the effect of different number of silicon nanowire transducer channels, in other word single, double, and triple channels towards the performance of field-effect transistor-based biosensor through simulation tool. These silicon nanowire field-effect transistor biosensors were designed and simulated in device simulation modelling tool, Silvaco ATLAS with fixed length, width, and height of the silicon nanowire. Different negatively interface charge density values were applied on the transducer channels’ surface of the biosensors to represent as detected target biomolecules that will bind onto the surface of the transducer regions. Based on the results, more negatively interface charges density values presented on the sensing channels had reduced the electron carrier accumulation at the channel’s interface, therefore, reduced drain current flow between the source and drain terminal. With the increase number of the transducer channels, significant change in drain current for every applied negatively interface charges became more apparent and increased the sensitivity of the biosensor. The triple transducer channels silicon nanowire field-effect transistor biosensor had demonstrated highest sensitivity, that is 2.83 µA/e∙cm2, which indicates it has better response for the detection of interface charges, thus increases chances for transducer channels reaction to the target biomolecules during testing or diagnosis.