The laboratory diagnosis of bovine tuberculosis, based on classical methods of culture isolation and biochemical identification of mycobacteria, is time-consuming, resource-intensive, and does not always allow for precise intraspecies differentiation. The aim of the study was to develop and test a rapid method for species identification of bacteria of the genus Mycobacterium based on the analysis of whole-cell ultraviolet resonance Raman spectroscopy (UV RRS) spectra. The study of 29 strains demonstrated that the use of two excitation wavelengths (248 nm and 251 nm) enables selective analysis of the protein-lipid complex and nucleic acids of the cell. For quantitative identification, two highly specific spectral indices were proposed: I 1660 /I 1615 (C = O/Tyr), sensitive to lipid composition and protein secondary structure, and I 1480 /I 1309 (G/A), correlating with genomic GC content. The method provides reliable differentiation, including within the M. tuberculosis complex (e.g., between M. tuberculosis and M. bovis), while maintaining accuracy after gentle pathogen inactivation and regardless of the culture growth phase, which opens prospects for its implementation in veterinary laboratory practice.
mycobacteria, ultraviolet resonance raman spectroscopy, species identification, macromolecular composition, multivariate statistical analysis, mycobacterium bovis, tuberculosis
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