Raman Spectroscopic Algorithms for Assessing Virulence in Oral Candidiasis: The Fight-or-Flight Response

This study aimed to test the effectiveness of Raman spectroscopy in the characterization of the degrees of physiological stress and virulence in clinical swab samples collected from patients affected by oral candidiasis. Raman experiments were conducted on a series of eight isolates, both in an as-collected state and after biofilm purification followed by 3 days of culture. The outputs were matched to optical microscopy observations and the results of conventional chromogenic medium assays. A statistically significant series of ten Raman spectra were collected for each clinical sample, and their averages were examined and interpreted as multiomic snapshots for <i>albicans</i> and non-<i>albicans</i> species. Spectroscopic analyses based on selected Raman parameters previously developed for standard <i>Candida</i> samples revealed an extreme structural complexity for all of the clinical samples, which arose from the concurrent presence of a variety of biofilms and commensal bacteria in the samples, as well as a number of other biochemical circumstances affecting the cells in their physiological stress state. However, three Raman algorithms survived such complexity, which enabled insightful classifications of <i>Candida</i> cells from clinical samples, in terms of their physiological stress and morphogenic state, membrane permeability, and virulence. These three characteristics, in turn, converged into a seemingly "fight or flight" response of the <i>Candida</i> cells. Although yet preliminary, the present study points out criticalities and proposes solutions regarding the potential utility of Raman spectroscopy in fast bedside analyses of surveillance samples.