The Application of Matrix-Assisted Laser Desorption/Ionisation Time of Flight Mass Spectrometry to Profile the Surface of Intact Bacterial Cells

Article English OPEN
Shah, Haroun N. ; Keys, Carrina J. ; Gharbia, Saheer E. ; Ralphson, Kathryn ; Trundle, Frank ; Brookhouse, Ian ; Claydon, Martin A. (2011)
  • Publisher: Microbial Ecology in Health and Disease
  • Journal: Microbial Ecology in Health and Disease (issn: 1651-2235, eissn: 1651-2235)
  • Related identifiers: doi: 10.3402/mehd.v12i4.8197

Matrix-Assisted Laser Desorption/lonisation Time of Flight Mass Spectrometry (MALDI-TOF-MS) as a tool for differentiating bacterial species was examined using reference strains representing gram-positive and gram-negative taxa. Initially, the effect of differences in medium composition on spectral profile was examined. The results indicated that growth on Columbia blood agar resulted in a larger spectrum of ionized residues and was therefore used for the cultivation of all strains in the rest of the study. The stability of the obtained mass spectral profiles against differences in batch and media processing suggested that no significant alterations to the profiles occurred in response to changes in media sources. The established conditions from these initial experiments were used to standardize subsequent experiments. The MALDI-TOF-MS profiles of 15 reference strains were compared and species characteristic markers were identified. The potential of using MALDI-TOF-MS as a tool for probing clonal diversity was examined using well characterized but clonally variable isolates of Bacteroides fragilis. Comparative analysis of the profiles of 20 strains revealed 5 clusters within the species but compared to other taxa such as Bacteroides merdae and Salmonella arizonae they form a closely related lineage. These results obtained strongly support the potential to use MALDI-TOF-MS as a tool for exploring bacterial surfaces for characteristic biomarkers and species-specific signatures.
  • References (8)

    1. Claydon MA, Shah HN, Borman P, Evason D, Gordon DB. Differentiation of biochemically inert, poorly characterised members of the genus Porphyromonas gingivalis by mass spectrometry of intact cells. Microbial Ecol. Health Dis. 1998; 10: 124.

    2. Claydon MA, Davey SN, Edward-Jones V, Gordon DB. The rapid identi cation of microorganisms using mass spectrometry. Nature Biotechnol. 1996; 14: 1584 - 6.

    3. Demirev PA, Ho Y-P, Ryzhov V, Fenselau C. Microorganism Identi cation by Mass Spectrometry and Protein Database Searches. Analyt. Chem. 1999; 71: 2732 - 8.

    4. Hillenkamp F, Karas M. Matrix-Assisted Laser Desorption:Ionization Mass Spectrometry of Lipopolymers. Analyt. Chem. 1991; 63: 1193 -202.

    5. Holland RD, Wilkes JG, Ra i F, Sutherland JB, Persons CC, Vorhees KJ, Lay Jr. JO. Rapid identi cation of Intact Whole Bacteria Based on Spectral Patterns using Matrix-assisted Laser Desorption:Ionization with Time-of-Flight Mass Spectrometry. Rapid Commun. Mass Spectrom. 1996; 10: 1227 - 32.

    6. Holland RD, Duffy CR, Raf i F, Sutherland JB, Heinze TM, Holder CL, Vorhees KJ, Lay Jr. JO. Identi cation of Bacterial Proteins Observed in MALDI TOF Mass Spectra from Whole Cells. Analyt. Chem. 1999; 71: 3226 - 30.

    7. Liang X, Zheng K, Quian MG, Lubman DM. Determination of Bacterial Protein Pro les by Matrix-assisted Laser Desorption: Ionization Mass Spectrometry with High-Performance Liquid Chromatography. Rapid Commun. Mass Spectrom. 1996; 10: 1219 - 26.

    8. Shah HN, Gharbia SE, Duerden BI. Bacteroides, Prevotella and Porphyromonas. In: Collier L, Balows A, Sussman M, eds. Topley and Wilson's Microbiology and Microbial Infections. London: Arnold, 1998: 1305 - 30.

  • Metrics
    No metrics available
Share - Bookmark