Cancer is one of the main causes of death in today's world, therefore, extending fast and reliable approaches for early monitoring of cancer cells have become one of the great challenges for scientists. This work reports exploiting a fast single frequency impedance strategy coupled to a new constructed composite for the quantitative detection of the human chronic myelogenous leukemia (CML) cell lines, K562. The platform used for the fabrication of the cytosensor is based on an electrospun poly[ethersulfone] (PES) sub-layer ready for improving by the upper nanodimension layers comprised of gold nanoparticles and multiwall carbon nanotubes (AuNPs/MWCNTs). The binding of K562 cancer cells with the pre-immobilized capture ssDNA was successfully transduced by Faradaic impedance spectroscopy (FIS). To diminish the possible nonspecific impedance changes that may cause some uncertainties in FIS signal, and to considerably decrease the time of the extended method, a fast single frequency measurement (SFM) strategy was tested based on recording total impedance |Z| in different individual frequencies, the methodology that was succeeded to recognize the target cells in measurement times as low as several seconds. Under the optimum conditions, the assessed signals were proportional to the population of K562 cells from 10(2) to 10(7) cells mL(-1) with a calculated detection limit of 60 cells mL(-1). The proposed biosensing-device also demonstrated high stability and reproducibility, which was able to offer great promise for the quantitative assay of K562 in routine analyses. The results of real sample analyses introduced the proposed device as alternative tool for CIVIL K562 cell detection in real biological samples. (C) 2018 Elsevier Ltd. All rights reserved.