Characterization and Differentiation of on Dixon's Agar Using Raman Spectroscopy.

, an emerging multidrug-resistant fungal pathogen, poses significant challenges in healthcare settings due to its high misidentification rate and resilience to treatments. Despite advancements in diagnostic tools, a gap remains in rapid, cost-effective identification methods that can differentiate from other species, particularly on non-standard culture media. We used Raman spectroscopy to characterize grown on modified Dixon's agar (mDixon) and differentiated it from and . Key Raman spectral markers at 1171 cm and 1452 cm, linked to mannan and β-glucan composition, differentiated into two subgroups, A and B. Despite the spectral similarities of groups A and B with and , respectively, all species were distinguishable through principal component analysis (PCA). Additionally, this study is the first to demonstrate the distinct spectral signature of mDixon agar, achieved through spatially offset Raman spectroscopy (SORS), which enables accurate discrimination between the culture medium and fungal samples. The observed inter-individual variability within , coupled with the spectral overlap between subgroups and other species, highlights a major challenge in differentiating closely related fungi due to their similar molecular composition. Enhancements in spectral resolution and further fluorescence minimization from the culture medium are needed to reliably detect the subtle biochemical differences within these species. Despite these challenges, the results underscore the potential of Raman spectroscopy as a real-time, non-destructive, and complementary tool for fungal pathogen identification.