Microorganisms assigned as Cronobacter are Gram-negative, facultatively anaerobic, bacteria widely distributed in nature, home environments, and hospitals. They can also be detected in foods, milk powder, and powdered infant formula (PIF). Additionally, as an opportunistic pathogen, Cronobacter may cause serious infections, sometimes leading to the death of neonates and infants. Thus, it is essential to test food products for the presence of Cronobacter spp. The currently used standard described in ISO 22964:2017 is a laborious method that could be easily replaced by surface-enhanced Raman scattering (SERS). Here, we demonstrate that SERS allows the identification of food-borne bacteria belonging to Cronobacter spp. based on their SERS spectra. For this purpose, twenty-six Cronobacter strains from different food samples were analyzed. Additionally, it was shown that it is possible to differentiate them from other closely related pathogens such as Salmonella enterica subsp. enterica, Escherichia coli, or Enterobacter spp. The SERS results were supported by principal component analysis (PCA), as well as and sequencing of 16S rRNA, rpoB and fusA genes. Last but not least, it was demonstrated that the cells of Cronobacter sakazakii may be easily separated from PIF using an appropriate filter, microfluidic chip, and dielectrophoresis (DEP) technique.