Proteomic Profiling of Intact Mycobacteria by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Details

• Personal Author:
  Hettick JM ; Kashon ML ; Mazurek GH ; Siege PD ; Simpson JP ; Weissman, David N.
• Description:
  Current methods for the identification of mycobacteria in culture are time-consuming, requiring as long as 12 weeks for positive identification. One potential approach to rapid mycobacterial identification is to utilize proteomic profiling of cultures by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In this report, we have applied MALDI-TOF MS to proteomic profiling of cultured microorganisms representing six species of the genus Mycobacterium. We find that analysis of acetonitrile/trifluoroacetic acid cellular extracts produces data similar to that of the analysis of deposited whole cells, while minimizing human contact with the microorganisms and rendering them nonviable. A matrix composition of alpha-cyano-4-hydroxycinnamic acid with fructose yields highly reproducible MALDI-TOF spectra. Statistical analysis of MALDI-TOF MS data allows differentiation of each individual mycobacterial species on the basis of unique mass fingerprints. The methodology allows identification of a number of unique (potentially diagnostic) biomarkers as targets for protein identification by MS/MS experiments. In addition, we observe a number of signals common to all mycobacterial species studied by MALDI-TOF MS, which may be genus-specific biomarkers. The potentially genus-specific biomarkers occur at low mass (<2 kDa) and are likely to be lipids and cell wall components such as mycolic acids. This study demonstrates the potential for mass spectrometry-based identification/classification of mycobacteria. [Description provided by NIOSH]
• Subjects:
  Acetone Biomarkers Communicable Diseases Mass Spectrometry Microbiology Occupational Health Safety
• Keywords:
  Acetones Bacteria Disaster-prevention Diseases Infectious-diseases Ionization Lasers Mass-spectrometry Microorganisms Statistical-analysis
• ISSN:
  0003-2700
• Document Type:
  Text
• Genre:
  Journal Article
• Place as Subject:
  Georgia ; OSHA Region 3 ; OSHA Region 4 ; 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:
  76
• Issue:
  19
• NIOSHTIC Number:
  nn:20025523
• Citation:
  Anal Chem 2004 Oct; 76(19):5769-5776
• Contact Point Address:
  National Institute for Occupational Safety and Health, Centers for Disease Control and Prevention, 1095 Willowdale Road, Morgantown, WV 26505
• Email:
  ayf2@cdc.gov
• CAS Registry Number:
  Acetonitrile (CAS RN 75-05-8)
• Federal Fiscal Year:
  2005
• NORA Priority Area:
  Disease and Injury: Infectious Diseases
• Peer Reviewed:
  True
• Source Full Name:
  Analytical Chemistry
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:11bad5305beae1e7c7e5fead00622e704d0f8f7bdb90e1e9e9a7eaeac60fd1f3300376162394b5c955e8de15bac42a1d33bb0f57e8cb2ea25daec355739e6894
• Download URL:
  https://stacks.cdc.gov/view/cdc/196849/cdc_196849_DS1.pdf
• File Type:
  [PDF - 182.55 KB ]