Reagent-Free Identification of Clinical Yeasts by Use of Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy

Abstract

Invasive fungal infections by opportunistic yeasts have increased concomitantly with the growth of an immunocompromised patient population. Misidentification of yeasts can lead to inappropriate antifungal treatment and complications. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy is a promising method for rapid and accurate identification of microorganisms. ATR-FTIR spectroscopy is a standalone, inexpensive, reagent-free technique that provides results within minutes after initial culture. In this study, a comprehensive spectral reference database of 65 clinically relevant yeast species was constructed and tested prospectively on spectra recorded (from colonies taken from culture plates) for 318 routine yeasts isolated from various body fluids and specimens received from 38 microbiology laboratories over a 4-month period in our clinical laboratory. ATR-FTIR spectroscopy attained comparable identification performance with matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS). In a preliminary validation of the ATR-FTIR method, correct identification rates of 100% and 95.6% at the genus and species levels, respectively, were achieved, with 3.5% unidentified and 0.9% misidentified. By expanding the number of spectra in the spectral reference database for species for which isolates could not be identified or had been misidentified, we were able to improve identification at the species level to 99.7%. Thus, ATR-FTIR spectroscopy provides a new standalone method that can rival MALDI-TOF MS for the accurate identification of a broad range of medically important yeasts. The simplicity of the ATR-FTIR spectroscopy workflow favors its use in clinical laboratories for timely and low-cost identification of life-threatening yeast strains for appropriate treatment.

Automated characterisation of neutrophil activation phenotypes in ex vivo human Candida blood infections

“…In particular, modern label-free methods have a promising potential in the future, among which various types of spectral imaging including Raman spectroscopy, Fourier transform infrared (FTIR) spectroscopy, or matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) providing information about the molecular composition of individual cells. Even though the efficiency of these spectroscopy techniques has been demonstrated for fungal cultures (e.g., [61] , [62] , [63] ), limitations for their application in the rapid identification of pathogens in human blood remain. In particular, methods based on cell cultivation require more than 24–48 h, which can lead to fatal delays in initiating pathogen-specific therapy.…”