Abstract
Acinetobacter baumannii (A. baumannii) is an emerging Gram-negative nosocomial pathogen responsible for infection on a global scale. It has the ability to develop biofilms on different surfaces, especially abiotic surfaces, which is considered a major contributor of its pathogenicity. Surface-enhanced Raman spectroscopy (SERS) holds great potential as an effective method for identifying and characterizing the biochemical composition of biofilm-forming species. In this study, cell mass samples from different strains of A. baumannii, categorized based on their biofilm-forming ability (strong, medium and non-biofilm forming) using a 96-well microtiter plate assay (MTP), were analyzed by SERS. The identified spectral features of the SERS spectra were used to characterize bacterial strains capable of producing biofilms. Silver nanoparticles (Ag-NPs) served as the SERS substrate to differentiate biofilm-forming strains of A. baumannii. Chemometric tools, such as principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA), were employed for the classification and differentiation of SERS spectra from bacterial strains with varying biofilm-producing capacities, achieving 100% sensitivity, 94.3% specificity, and an area under the curve (AUC) value of 0.81 through Monte Carlo cross-validation. Furthermore, K-fold (Leave-K-out cross-validation (LKOCV)) was applied to verify the robustness of the PLS-DA model, and the AUC value was found to be 0.90, with a sensitivity of 100% and specificity of 98%. These results demonstrate that the PLS-DA model is highly effective for the differentiation and classification of bacterial strains with varying capacities for biofilm production.