In the present work, for the first time we takes the advantages of chitosan-Nafion (Chit-Naf) composite as a highly conductive surface platform and a novel CNT-based signal amplification strategy to develop a lable-free impedimetricaptamer-based sensor for highly sensitive detection of As(III). The electrochemical impedance spectroscopy (EIS) investigations surprisingly revealed that the glassy carbon electrode (GC) electrode modified with Chit-Naf composite had higher electron transfer kinetics compared the bare GC, GC/Naf and GC/Chit electrodes, which promises a great potential as an efficient platform in construction of biosensing assays. In this work, we employed a signal amplification strategy based on carbon nanotube-bovine serum albumin (CNT-BSA) hybrid system, by which sensitivity and detection limit of the aptasensor for the detection of As(III) were obtained to be 100.82 Ω nM-1 and a of 74 pM, respectively. This protocol provided one of the lowest limits of detection for As(III) on aptamer-based electrodes recently described in the literature. Moreover, the change of the optical absorptive properties of CNTs upon biorecognition interactions provides a way to detect the biorecognition process and thus allowed us to design an optical As(III) aptasensor using the UV-Vis spectroscopic method. The discrimination capability of the fabricated aptasensor for recognizing As(III) in the presence of other metal ions and a complex matrix of waste water samples was successfully investigated. This protocol provided a new method for sensitive detection of As(III) with considerable advantages in terms of reproducibility, selectivity, being mediator free and regenerability of the sensing interface.