Microwell plate-integrated glass-silver nanoforest SERS platform for high-throughput multiplexed detection of foodborne pathogens

초록

Rapid and accurate detection of foodborne pathogens such as Escherichia coli (E. coli), Salmonella, and Staphylococcus aureus (S. aureus) is essential for securing public health; however, conventional detection methods such as polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), and matrix-assisted laser desorption/ionization - time-of-flight (MALDI-TOF) are limited by their complex operation. Surface-enhanced Raman scattering (SERS) allows for sensitive, label-free detection of microorganisms; however, existing platforms mostly rely on unstable colloidal nanoparticles or opaque substrates that limit imaging and highthroughput analysis. We present a high-throughput SERS platform using a transparent glass-silver nanoforest (GSNF) substrate integrated with a custom-designed Raman reader. The GSNF enables strong signal enhancement with low autofluorescence, while a 3D-printed sensor array and the automated reader allow multiplexed and spatially resolved measurements. The system exhibits excellent reproducibility with rhodamine 6G and successfully differentiates foodborne pathogens using principal component analysis. It can also identify speciesspecific peaks in mixed bacterial samples, demonstrating its potential for scalable and practical multiplexed diagnostics. Furthermore, the system successfully detected E. coli in chicken breast extract, highlighting its applicability to real food matrices and its promise for practical food safety monitoring.

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