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Species identification of Aspergillus, Fusarium and Mucorales with direct surface analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.
Clin Microbiol Infect. 2012 May; 18(5):475-84.CM

Abstract

Accurate species discrimination of filamentous fungi is essential, because some species have specific antifungal susceptibility patterns, and misidentification may result in inappropriate therapy. We evaluated matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for species identification through direct surface analysis of the fungal culture. By use of culture collection strains representing 55 species of Aspergillus, Fusarium and Mucorales, a reference database was established for MALDI-TOF MS-based species identification according to the manufacturer's recommendations for microflex measurements and MALDI BioTyper 2.0 software. The profiles of young and mature colonies were analysed for each of the reference strains, and species-specific spectral fingerprints were obtained. To evaluate the database, 103 blind-coded fungal isolates collected in the routine clinical microbiology laboratory were tested. As a reference method for species designation, multilocus sequencing was used. Eighty-five isolates were unequivocally identified to the species level (≥99% sequence similarity); 18 isolates producing ambiguous results at this threshold were initially rated as identified to the genus level only. Further molecular analysis definitively assigned these isolates to the species Aspergillus oryzae (17 isolates) and Aspergillus flavus (one isolate), concordant with the MALDI-TOF MS results. Excluding nine isolates that belong to the fungal species not included in our reference database, 91 (96.8%) of 94 isolates were identified by MALDI-TOF MS to the species level, in agreement with the results of the reference method; three isolates were identified to the genus level. In conclusion, MALDI-TOF MS is suitable for the routine identification of filamentous fungi in a medical microbiology laboratory.

Authors+Show Affiliations

Institute of Microbiology, Università Cattolica del Sacro Cuore, Largo F. Vito 1, Rome, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21883662

Citation

De Carolis, E, et al. "Species Identification of Aspergillus, Fusarium and Mucorales With Direct Surface Analysis By Matrix-assisted Laser Desorption Ionization Time-of-flight Mass Spectrometry." Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases, vol. 18, no. 5, 2012, pp. 475-84.
De Carolis E, Posteraro B, Lass-Flörl C, et al. Species identification of Aspergillus, Fusarium and Mucorales with direct surface analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clin Microbiol Infect. 2012;18(5):475-84.
De Carolis, E., Posteraro, B., Lass-Flörl, C., Vella, A., Florio, A. R., Torelli, R., Girmenia, C., Colozza, C., Tortorano, A. M., Sanguinetti, M., & Fadda, G. (2012). Species identification of Aspergillus, Fusarium and Mucorales with direct surface analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases, 18(5), 475-84. https://doi.org/10.1111/j.1469-0691.2011.03599.x
De Carolis E, et al. Species Identification of Aspergillus, Fusarium and Mucorales With Direct Surface Analysis By Matrix-assisted Laser Desorption Ionization Time-of-flight Mass Spectrometry. Clin Microbiol Infect. 2012;18(5):475-84. PubMed PMID: 21883662.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Species identification of Aspergillus, Fusarium and Mucorales with direct surface analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. AU - De Carolis,E, AU - Posteraro,B, AU - Lass-Flörl,C, AU - Vella,A, AU - Florio,A R, AU - Torelli,R, AU - Girmenia,C, AU - Colozza,C, AU - Tortorano,A M, AU - Sanguinetti,M, AU - Fadda,G, Y1 - 2011/08/29/ PY - 2011/9/3/entrez PY - 2011/9/3/pubmed PY - 2012/8/7/medline SP - 475 EP - 84 JF - Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases JO - Clin Microbiol Infect VL - 18 IS - 5 N2 - Accurate species discrimination of filamentous fungi is essential, because some species have specific antifungal susceptibility patterns, and misidentification may result in inappropriate therapy. We evaluated matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for species identification through direct surface analysis of the fungal culture. By use of culture collection strains representing 55 species of Aspergillus, Fusarium and Mucorales, a reference database was established for MALDI-TOF MS-based species identification according to the manufacturer's recommendations for microflex measurements and MALDI BioTyper 2.0 software. The profiles of young and mature colonies were analysed for each of the reference strains, and species-specific spectral fingerprints were obtained. To evaluate the database, 103 blind-coded fungal isolates collected in the routine clinical microbiology laboratory were tested. As a reference method for species designation, multilocus sequencing was used. Eighty-five isolates were unequivocally identified to the species level (≥99% sequence similarity); 18 isolates producing ambiguous results at this threshold were initially rated as identified to the genus level only. Further molecular analysis definitively assigned these isolates to the species Aspergillus oryzae (17 isolates) and Aspergillus flavus (one isolate), concordant with the MALDI-TOF MS results. Excluding nine isolates that belong to the fungal species not included in our reference database, 91 (96.8%) of 94 isolates were identified by MALDI-TOF MS to the species level, in agreement with the results of the reference method; three isolates were identified to the genus level. In conclusion, MALDI-TOF MS is suitable for the routine identification of filamentous fungi in a medical microbiology laboratory. SN - 1469-0691 UR - https://www.unboundmedicine.com/medline/citation/21883662/Species_identification_of_Aspergillus_Fusarium_and_Mucorales_with_direct_surface_analysis_by_matrix_assisted_laser_desorption_ionization_time_of_flight_mass_spectrometry_ DB - PRIME DP - Unbound Medicine ER -