A reusable QCM biosensor with stable antifouling nano-coating for on-site reagent-free rapid detection of E. coli O157:H7 in food products
Michala Forinová, Alina Pilipenco, N. Scott Lynn, Radka Obořilová, Hana Šimečková, Markéta Vrabcová, Monika Spasovová, Rachael L. Jack, Petr Horák, Milan Houška, Petr Skládal, Petr Šedivák, Zdeněk Farka, Hana Vaisocherová‐Lísalová
Abstract
Numerous biosensors have shown exceptional analytical performance under laboratory conditions, yet only a few are capable of on-site use with complex, non-model samples while exhibiting reliable analytical performance. Here, we present a new portable biosensor for the rapid (30 min) and accurate detection of bacterial agents in “real-world” food samples, which are originally in either solid or liquid form. The biosensor combines well-established quartz crystal microbalance (QCM) technology, with innovative terpolymer brush nano-coatings on the sensing surface to efficiently reduce non-specific fouling from food samples. Following reagent-free sample preparation, where solid food samples are homogenized, we validated the sensor's detection capabilities on native pathogenic Escherichia coli O157:H7 (E. coli O157:H7) in hamburgers, Czech dumplings, and milk. We achieved limits of detection (LOD), as low as 7.5 × 102 CFU/mL in milk, a value approaching fundamental QCM limits, using a simple direct detection assay format. The biosensor's exceptional reusability was demonstrated through 60 sequential hamburger sample injections, resulting in only a minor LOD shift toward the end of series. A 10-min sonication treatment during sample preparation significantly enhanced sensitivity for E. coli O157:H7 in hamburgers and dumplings, yielding LODs as low as 3.1 × 10³ CFU/mL and 2.6 × 10⁴ CFU/mL, respectively. For on-site analysis, we integrated the nano-coated sensing chip into a custom-built four-channel portable QCM biosensor with an optimized microfluidic system, which can be produced on a scale suitable for practical deployment.