title: Rapid identification of 6328 isolates of pathogenic yeasts using MALDI-ToF MS and a simplified, rapid extraction procedure that is compatible with the Bruker Biotyper platform and database.
abstract: Rapid and accurate identification of yeast isolates from clinical samples is essential, given their innately variable antifungal susceptibility profiles, and the proposal of species-specific antifungal susceptibility interpretive breakpoints. Here we have evaluated the utility of MALDI-ToF MS analysis for the identification of clinical isolates of pathogenic yeasts. A simplified, rapid extraction method, developed in our laboratory, was applied to 6343 isolates encompassing 71 different yeast species, which were then subjected to MALDI-ToF MS analysis using a Bruker Microflex and the resulting spectra were assessed using the supplied Bruker database. In total, 6328/6343 (99.8%) of isolates were correctly identified by MALDI-ToF MS. Our simplified extraction protocol allowed the correct identification of 93.6% of isolates, without the need for laborious full extraction, and a further 394 (6.2%) of isolates could be identified after full extraction. Clinically relevant identifications with both extraction methods were achieved using the supplied Bruker database and did not require the generation of bespoke, in-house databases created using profiles obtained with the adapted extraction method. In fact, the mean LogScores obtained using our method were as robust as those obtained using the recommended, published full extraction procedures. However, an in-house database can provide a useful additional identification tool for unusual or rarely encountered organisms. Finally, the proposed methodology allowed the correct identification of over 75% of isolates directly from the initial cultures referred to our laboratory, without the requirement for additional sub-culture on standardised mycological media.
authors.affiliation: UK National Mycology Reference Laboratory, Public Health England, Bristol, United Kingdom Mark.Fraser@uhBristol.nhs.uk.
authors.forename: Mark
authors.initials: M
authors.lastname: Fraser
authors.affiliation: UK National Mycology Reference Laboratory, Public Health England, Bristol, United Kingdom.
authors.forename: Zoe
authors.initials: Z
authors.lastname: Brown
authors.affiliation: UK National Mycology Reference Laboratory, Public Health England, Bristol, United Kingdom.
authors.forename: Marian
authors.initials: M
authors.lastname: Houldsworth
authors.affiliation: UK National Mycology Reference Laboratory, Public Health England, Bristol, United Kingdom.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Borman
authors.affiliation: UK National Mycology Reference Laboratory, Public Health England, Bristol, United Kingdom.
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Johnson
mesh_terms: D005656:Fungal Proteins / Q000302:isolation & purification*; D006801:Humans; D008828:Microbiological Techniques / Q000379:methods*; D009181:Mycoses / Q000382:microbiology*; D013048:Specimen Handling / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013997:Time Factors; D015003:Yeasts / Q000737:chemistry* / Q000145:classification* / Q000302:isolation & purification
chemical_list: D005656:Fungal Proteins
keywords: Candida species; MALDI-ToF MS; pathogenic yeasts; rapid extraction method; rapid identification