Sensory protocols for evaluation of DNA distortion due to exposure to various harmful chemicals and environments in living cells are needed for research and clinical investigations. Here, a design of non-metal sensory (NMS) electrode was built by using boron-doped carbon spherules for detection of DNA nucleobases, namely, guanine (Gu), adenine (Ad), and thymine (Th) in living cells. The key-electrode based nanoscale NMS structures lead to voids with a facile diffusion, and strong binding events of the DNA nucleobases. Furthermore, the NMS geometric structures would significantly create electrode surfaces with numerous centrally active sites, curvature topographies, and anisotropic spherules. The NMS shows potential as sensitive protocol for DNA-nucleobases in living cells exposed to oxidative stresses. In one-step signaling assay, NMS shows high signaling transduction of Gu-, Ad-, and Th-DNA nucleobases targets with ultra-sensitive and low detection limits of 3.0, 0.36, and 0.34 nM, respectively, and a wide linear range of up to 1 μM. The NMS design and protocol show evidence of the role of surface construction features and B-atoms incorporated into the graphitic carbon network for creating abundant active sites with facile electron diffusion and heavily target loads along with within-/out-plane circular spheres. Indeed NMS, with spherule-rich interstitial surfaces can be used for sensitive and selective evaluation of damaged-DNA to various dysfunctional metabolism in the human body.