Identifying Fungal Isolates Using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Details

• Personal Author:
  Beezhold, Donald H. ; Blanchere, Francoise M. ; Buskirk A ; Green, Brett J. ; Hettick JM ; Janotka E ; Kashon ML ; Schmechel D ; Siegel, Paul D. ; Slaven, James E.
• Description:
  RATIONALE: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a powerful tool for phenotyping microorganisms. The purpose of this study was to utilize MALDI-TOF MS to determine whether the mass spectra of individual fungal species produced specific m/z values that could be used to discriminate between species. METHODS: Sixty two different fungal isolates were grown for 14 days on malt extract agar. Conidia and hyphae were extracted in 250 mg of zirconium beads in 1mL 50/50 acetonitrile/4% trifluoroacetic acid by three 1 min bead beating cycles. The resultant extract was centrifuged at 14,500 rpm for 10 minutes and the supernatant analyzed. Samples were mixed 1:1 with 10 mg/mL alpha-cyano-4-hydroxycinnamic acid. A 1 mL aliquot was spotted on a gold target plate and allowed to air dry before MALDI-TOF MS analysis. RESULTS: MALDI-TOF MS mass spectra of fungal isolates contained numerous peaks within the 1,000 - 40,000 mass-to-charge (m/z) range. Statistical discriminative analysis of mass spectra resulted in the ability to unambiguously differentiate fungal species. The mass spectra of closely related species are often similar; however, these may still be distinguished by species-specific m/z values and/or differential relative abundance of shared m/z values. CONCLUSIONS: This work demonstrates the potential utility of MALDI-TOF MS for the objective species identification of both clinically and environmentally relevant fungal isolates. [Description provided by NIOSH]
• Subjects:
  Fungi Hypersensitivity Mass Spectrometry Microbiology Occupational Health Radiation Safety
• Keywords:
  Allergens Allergic-disorders Allergic-reactions Analytical-processes Fungal-diseases Fungal-infections Immune-system-disorders Ionization Laser-radiation Lasers Mass-spectrometry Microscopic-analysis Sample-preparation

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• ISSN:
  0091-6749
• Document Type:
  Text
• Genre:
  Abstract or Summary ; Proceedings
• Place as Subject:
  OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  HELD - Health Effects Laboratory Division
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Volume:
  121
• Issue:
  2
• NIOSHTIC Number:
  nn:20033489
• Citation:
  J Allergy Clin Immunol 2008 Feb; 121(2)(Suppl 1):S270
• Federal Fiscal Year:
  2008
• Peer Reviewed:
  False
• Source Full Name:
  Journal of Allergy and Clinical Immunology
• Supplement:
  1
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:0763477e8dcca0682723a8d9c94d51a8e8f7400a8f2f793bcf96fc783ae19f0e138846d6bedfae05eae39bae52a39b4eff827404038681b22a1a381f2b37009e
• Download URL:
  https://stacks.cdc.gov/view/cdc/189039/cdc_189039_DS1.pdf
• File Type:
  [PDF - 52.34 KB ]