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One‐dimensional zinc oxide nanomaterials (1DZnO) have emerged as promising, cost‐effective nanoplatforms with adjustable properties suitable for electrochemical and optical biosensing applications. In this work, modifications in the inherent photoluminescent response of 1DZnO are harnessed to develop a novel immunosensor tailored for detecting enteropathogenic Escherichia coli. This nanobiosensor demonstrates a modulation in photoluminescence signal, effectively responsive to analyte concentrations ranging from 1 × 102 to 1 × 108 CFU mL−1, with direct visualization of targeted bacterial cells over 1DZnO structures through scanning electron microscopy. The conceptualization of this nanobiosensor is focused on a real‐time contact strategy that can significantly reduce processing and response times for pathogen detection, prospected for emergency scenarios. With this aim, the detection process unfolds in real time, with a mere 5–10 s interaction time, corroborated by the standard polymerase chain reaction approach. This synergistic validation underscores the reliability and precision of the developed biosensor. Notably, the utility of 1DZnO nanoplatforms extends beyond the realm of enteropathogenic E. coli, as the biosensing performance exhibited here holds promise for analogous applications involving other medically pertinent pathogens. This study paves the way for the broader implementation of 1DZnO‐based biosensors in medical diagnostics, offering rapid, sensitive, and real‐time detection capabilities. [ABSTRACT FROM AUTHOR]