Rapid Identification of Foodborne Pathogens in Limited Resources Settings Using a Handheld Raman Spectroscopy Device

Featured Application Here, we report a practical and precise method for the identification of foodborne pathogenic bacteria using a Raman handheld device equipped with an orbital raster scan (ORS) technology that enables the system to generate a distinct fingerprint of bacteria suspended in media broth by applying principal component analysis (PCA) and support vector machine (SVM) as classifiers. This system relies on isolation from the source to ensure the generation of fingerprints of axenic cultures. It requires very little sample preparation and is significantly less costly than other Raman technologies. Thus, this system could be easily implemented in limited-resource settings. Rapid and precise methods to detect pathogens are paramount in ensuring food safety and selecting appropriate disinfection treatments. Raman spectrometry is a promising technology being investigated for detecting pathogens and achieving rapid, culture-free, and label-free methods. Nonetheless, previous Raman techniques require additional steps, including the preparation of slides that could introduce significant variability. In this study, we investigated the capability of a Raman handheld device for rapid identification of monocultures of Listeria monocytogenes, Salmonella Typhimurium, Escherichia coli O157:H7, and Staphylococcus aureus, and the combination of co-cultures in BHI broth suspension by utilising principal component analysis (PCA) and support vector machine (SVM) classification of Raman spectra. The detection method accurately identified monocultures (0.93 +/- 0.20), achieving good discrimination after 24 h of bacterial growth. However, the PCA-SVM system was less accurate for classifying co-cultures (0.67 +/- 0.35). These results show that this method requires an isolation step followed by biomass enrichment (>8 log(10) CFU/mL) for accurate identification. The advantage of this technology is its simplicity and low-cost preparation, achieving high accuracy in monocultures in a shorter time than conventional culture-dependent methods.