Identification of Staphylococcus aureus, Enterococcus faecium, Klebsiella pneumoniae, Pseudomonas aeruginosa and...

Rapid and accurate identification of bacteria is required in order to develop effective treatment strategies. Traditional culture-based methods are time-consuming, while MALDI-TOF MS is expensive. The Raman spectroscopy, due to its relatively cost-effectiveness, offers a promising alternative for bacterial identification. However, its clinical utility still requires further validation.

In this study, the artificial intelligent Raman detection and identification system (AIRDIS) was implemented to identify bacterial species, including Staphylococcus aureus (n = 1290), Enterococcus faecium (n = 1020), Klebsiella pneumoniae (n = 1366), Pseudomonas aeruginosa (n = 1067), and Acinetobacter baumannii (n = 811). Raman spectra were collected, preprocessed, and analyzed by machine learning (ML).

After training on 24,420 Raman spectra from 1221 isolates and testing on 4333 isolates, the AIRDIS demonstrated an area under the curve (AUC) of 0.99 for Gram classification, with accuracies of 97.64 % for Gram-positive bacteria and 98.86 % for Gram-negative bacteria. Spectral differences between Gram-positive and Gram-negative bacteria were linked to structural variations in their cell walls, such as peptidoglycan and lipopolysaccharides. At the species level, S. aureus, E. faecium, K. pneumoniae, P. aeruginosa, and A. baumannii were identified with high accuracy, ranging from 94.76 % to 96.88 %, with all species achieving an AUC of 0.99.

Validation with a large number of clinical isolates demonstrated Raman spectroscopy combined with ML excels in identification of five bacterial species associated with multidrug resistance. This finding confirms the clinical utility of the system while laying a solid foundation for the future development of antimicrobial resistance prediction models.