Characterisation of a SapYZU11@ZnFe2O4 biosensor reveals its mechanism for the rapid and sensitive colourimetric detection of viable Staphylococcus aureus in food matrices.

Although bacteriophage-based biosensors hold promise for detecting Staphylococcus aureus in food products in a timely, simple, and sensitive manner, the associated targeting mechanism of the biosensors remains unclear. Herein, a colourimetric biosensor SapYZU11@ZnFe2O4, based on a broad-spectrum S. aureus lytic phage SapYZU11 and a ZnFe2O4 nanozyme, was constructed, and its capacity to detect viable S. aureus in food was evaluated. Characterisation of SapYZU11@ZnFe2O4 revealed its effective immobilisation, outstanding biological activity, and peroxidase-like capability. The peroxidase activity of SapYZU11@ZnFe2O4 significantly decreased after the addition of S. aureus, potentially due to blockage of the nanozyme active sites. Moreover, SapYZU11@ZnFe2O4 can detect S. aureus from various sources and S. aureus isolates that phage SapYZU11 could not lyse. This may be facilitated by the adsorption of the special receptor-binding proteins on the phage tail fibre and wall teichoic acid receptors of S. aureus. Besides, SapYZU11@ZnFe2O4 exhibited remarkable sensitivity and specificity when employing colourimetric techniques to rapidly determine viable S. aureus counts in food samples, with a detection limit of 0.87 × 102 CFU/mL. Thus, SapYZU11@ZnFe2O4 has broad application prospects for the detection of viable S. aureus cells on food substrates.

Characterisation of a colourimetric biosensor SapYZUM13@Mn3O4-NH2 reveals the mechanisms underlying its rapid and sensitive detection of viable Staphylococcus aureus in food.

In this study, a colourimetric biosensor based on bacteriophage SapYZUM13 and an aminated Mn3O4 (Mn3O4-NH2) nanozyme was constructed and evaluated for its ability to detect Staphylococcus aureus in food. The biosensor had a detection time of 20 min, with a detection limit of 2 × 101 CFU/mL and recovery rate of 92.42-106.96%, indicating its high reliability and accuracy in detecting the food pathogen. Mechanistically, SapYZUM13@Mn3O4-NH2 exhibited oxidase-mimicking capability, producing O2•- free radicals which oxidise 3,3',5,5'-tetramethylbenzidine (TMB) to yield blue-coloured oxTMB. In the presence of S. aureus, the oxidase activity decreased remarkably owing to shielding of the nanozyme active sites. Moreover, SapYZUM13@Mn3O4-NH2 could detect viable S. aureus from various sources, likely because of the special receptor-binding proteins of SapYZUM13 adsorbing to the wall teichoic acids on the S. aureus cell surface. Thus, SapYZUM13@Mn3O4-NH2 has broad application prospects for the detection of viable S. aureus in various foods.