Development of a disposable paper-based potentiometric immunosensor for real-time detection of a foodborne pathogen

This work reports a new paper-based sensing platform and its application in a label-free potentiometric immunosensor for Salmonella typhimurium detection based on the blocking surface principle. A paper-based strip electrode was integrated with a filter paper pad which acted as a reservoir of the internal solution. The design offers a convenient platform for antibody immobilization and sampling, proving also that is a simple and affordable methodology to control an ionic flux through a polymer membrane. Two different immunosensing interfaces were assembled on the developed paper-strip electrode. The simplest interface relied on direct conjugation of the antibody to the polymer membrane and the second one resorted to an intermediate layer of a polyamidoamine dendrimer, with an ethylenediamine core from the fourth generation. Electrochemical impedance spectroscopy was used to assess the successive interface modification steps and the resulting analytical performance of both immunosensors was compared. For such, the potential shift derived from the blocking effect of the ionic flux caused by antigen-antibody conjugation was correlated with the logarithm of the Salmonella typhimurium concentration in the sample. In optimized conditions, a limit of detection of 5 cells mL-1 was achieved. As a proof-of-concept, the proposed method was applied to apple juice samples, demonstrating to be a suitable prototype to be used in real scenarios in useful time (<1 h assay).This work received financial support from the European Union and National Funds(FCT, Fundação para a Ciência e Tecnologia) through projects UID/QUI/50006/2019 and Norte-01-0145-FEDER-000011-RL1–QUALIFOOD. N.F.D Silva is grateful to FCT grant SFRH/BD/112414/2015, financed by POPH–QREN–Tipologia 4.1–FormaçãoAvançada,subsidizedbyFSEandMCTES.info:eu-repo/semantics/publishedVersio