title: MALDI-TOF MS for the identification of veterinary non-C. neoformans-C. gattii Cryptococcus spp. isolates from Italy.
abstract: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) offers an effective alternative to phenotypic and molecular methods for the rapid identification of microorganisms. Our aim in this study was to create an in-house library for a set of strains of nine uncommonly reported human and animal cryptococcal species, including Cryptococcus adeliensis, C. albidosimilis, C. albidus, C. aureus, C. carnescens, C. laurentii, C. magnus, C. victoriae and C. uniguttulatus, and to use this library to make timely and correct identifications using MALDI-TOF MS for use in routine laboratory diagnostics. Protein extracts obtained via the formic acid extraction method of 62 veterinary non-C. neoformans-C. gattii cryptococcal isolates were studied. The obtained mass spectra correctly grouped all 62 studied isolates according to species identification previously obtained by internal transcribe spacer sequence analysis. The in-house database was than exported and successfully uploaded to the Microflex LT (Maldi Biotyper; Bruker Daltonics) instrument at a different diagnostic laboratory in Italy. Scores >2.7 obtained from isolates reanalyzed in the latter laboratory supported the high reproducibility of the method. The possibility of creating and transferring an in-house library adds to the usefulness MALDI-TOF MS an important tool for the rapid and inexpensive identification of pathogenic and saprophytic fungi as required for differential diagnosis of human and animal mycoses.
authors.affiliation: Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro (Padua), Italy Department of Veterinary Medicine, University of Bari, Italy pdanesi@izsvenezie.it.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Danesi
authors.affiliation: Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro (Padua), Italy.
authors.forename: Ilenia
authors.initials: I
authors.lastname: Drigo
authors.affiliation: Department of Veterinary Medicine, University of Bari, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Iatta
authors.affiliation: Molecular Mycology Research Laboratory, Centre for Infectious Diseases and Microbiology, Sydney Medical School-Westmead Hospital, Marie Bashir Institute for Infectious Diseases and Biosecurity, University of Sydney, Westmead Millennium Institute, Sydney, Australia Grupo de Microbiología, Instituto Nacional de Salud, Bogotá, Colombia.
authors.forename: Carolina
authors.initials: C
authors.lastname: Firacative
authors.affiliation: Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro (Padua), Italy.
authors.forename: Gioia
authors.initials: G
authors.lastname: Capelli
authors.affiliation: Department of Veterinary Medicine, University of Bari, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Cafarchia
authors.affiliation: Molecular Mycology Research Laboratory, Centre for Infectious Diseases and Microbiology, Sydney Medical School-Westmead Hospital, Marie Bashir Institute for Infectious Diseases and Biosecurity, University of Sydney, Westmead Millennium Institute, Sydney, Australia.
authors.forename: Wieland
authors.initials: W
authors.lastname: Meyer
mesh_terms: D000818:Animals; D003453:Cryptococcosis / Q000175:diagnosis* / Q000382:microbiology*; D003454:Cryptococcus / Q000737:chemistry* / Q000145:classification* / Q000302:isolation & purification; D005656:Fungal Proteins / Q000032:analysis / Q000302:isolation & purification; D006801:Humans; D007558:Italy; D008828:Microbiological Techniques / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D005656:Fungal Proteins
keywords: MALDI-TOF; non–C. neoformans–C. gattii Cryptococcus species; veterinary isolates