According to WHO, cervical cancer is considered as one of the most frequently diagnosed cancers and the fourth main source of cancer death in women in 2020 worldwide. Hence, there is a need for development of cervical cancer screening with new rapid and cost-effective methods. Although there are few methods available for HPV identification, these techniques are less sensitive, time-consuming, and costly. An ultra-sensitive, selective, and label-free DNA-based impedimetric electrochemical genosensor is developed in this study to detect HPV-18 for cervical cancer. Electrochemical analysis was performed for the characterization of the sensing platform and for the detection of analyte. A single-stranded 25mer oligonucleotide DNA probe was immobilized onto a nitrogen-doped carbon nanodot-modified ITO electrode. Furthermore, the hybridization event was measured by testing the complementary single stranded DNA sequence in the samples. The sensor could distinguish between complementary as well as non-complementary sequences. Herein, impedance quantification demonstrated a limit of detection of 0.405 fM. The developed genosensor showed high selectivity toward HPV-18 in the clinical samples. This sensing platform can be considered as a rapid and selective method for the screening of HPV-18.