Sensitive, portable methods of detection for foodborne pathogens hold great significance for the early warning and prevention of foodborne diseases and environmental pollution. Restricted by a complicated matrix and limited signaling strategies, developing a ready-to-use sensing platform with ultrahigh sensitivity remains challenging. In this work, near-infrared (NIR) light-responsive AgBiS nanoflowers (NFs) and CuO nanocubes (NCs) were introduced to construct a novel target-induced photocurrent-polarity-switchable system and verified for the development of an all-in-one, ready-to-use photoelectrochemical (PEC) immunosensor. NIR-responsive n-type AgBiS NFs and p-type CuO NCs producing anodic and cathodic photocurrents were conjugated with monoclonal (MAb) and polyclonal antibodies (PAb), respectively. Using a sandwich-type immunocomplex bridged by O157:H7, an efficient photocurrent-polarity-switching PEC system was formed on a paper-based working electrode (PWE). Owing to the spatial separation of the photogenerated carriers and the elimination of false-positive/negative signals by the polarity-switchable photocurrent, the proposed NIR PEC immunoassay for O157:H7 exhibits a considerably low detection limit of 8 colony-forming units/milliliter (CFU/mL) with a linear range from 25 to 5 × 10 CFU/mL. The platform includes a PWE with an automatic cleaning function and a portable PEC analyzer with smartphone-compatible Bluetooth capability, thus achieving point-of-care testing of O157:H7. The sensor was applied to the analysis of pork samples artificially contaminated with O157:H7, and the detection results were in good agreement with the plate counting method, a gold standard in the field. This work aimed to investigate the photoelectric activity of the NIR-responsive p/n-type composites and to provide a new signal-reversal route for the construction of an all-in-one ready-to-use PEC immunosensor for the detection of low-concentration biomolecules.
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