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This work reports the development of an electrochemical immunosensor for rapid, specific and decentralized detection of the invasion-associated protein p60 secreted by Listeria monocytogenes, a life-threatening foodborne pathogen. A disposable screen-printed electrode was used as transducer surface and monoclonal and polyclonal antibodies that specifically recognize Listeria monocytogenes p60 protein and Listeria spp. p60 proteins, respectively, were used as the sandwich immuno-pair. The reaction was detected with the aid of an additional secondary antibody conjugated with the enzyme reporter (alkaline phosphatase) and using 3-indoxyl phosphate/silver ions as the mixture substrate. The analytical signal was acquired through the voltammetric stripping of the enzymatically deposited silver, which was directly correlated to p60 concentration in the sample. In optimized conditions, a limit of detection and quantification of 1.5 ng mL−1 and 5.1 ng mL−1 were achieved, respectively, in a useful time (<3 h). As proof-of-concept, the proposed immunosensor was successfully applied to spiked milk samples, demonstrating to be a suitable device for further use in real sample detection of Listeria monocytogenes in food products.
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•Listeria monocytogenes is a life-threatening foodborne pathogen.•Invasion-associated protein p60 is a virulence factor secreted by Listeria spp..•A disposable electrochemical immunosensor for Listeria monocytogenes p60 determination was developed.•The immunosensor achieved a limit of detection of 1.5 ng mL−1 in a useful time (<3 h).•The immunosensing approach accurately detected Listeria monocytogenes p60 in spiked milk samples.