Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

The rapid identification of intact microorganisms using mass spectrometry

Abstract

Antibiotic-resistant strains of bacteria continue to emerge, increasing the need for their fast and accurate identification. Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS), has become a prominent technique in biological mass spectrometry. We report the application of MALDI-TOF-MS for the identification of intact Gram-negative and Gram-positive microorganisms taken directly from culture. Analysis of bacteria from a single colony is possible, allowing the screening of mixed cultures. Sample preparation is simple and the analysis automated, providing spectra within minutes. The spectra obtained allow identification of microorganisms from different genera, different species, and from different strains of the same species. The procedure provides a unique mass spectral fingerprint of the microorganism, produced from desorbed components of the cell wall. Consistent data were obtained from subcultures grown for 3-day and 6-day periods, from the same cultures 1 day later and from fresh subcultures 2 months later.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Morse, A. 1995. Classification of bacteria, Chapter 3, pp. 35–43 in Medical microbiology, 20th ed. Jawetz, E., Melnick, J.L., and Adelberg, E.A. (eds.). Lange Medical, London.

    Google Scholar 

  2. Gallagher, R.T., Davey, S.N., and Derrick, P.J. 1996. pp. 329–364 in Biomedical applications of spectroscopy advances in spectroscopy, Vol. 25. Clark, R.J.H. and Hester, R.E. (eds.). John Wiley and Sons, Chichester, UK.

    Google Scholar 

  3. Fenselau, C. (ed.). 1994. Mass spectrometry for the characterization of microorganisms, ACS symposium series, Vol. 541. ACS, Washington, DC.

    Google Scholar 

  4. Heller, D.N., Fenselau, C., Cotter, R.J., Demirev, P., Olthoff, J.K., Honovich, J. et al. 1987. Mass spectral analysis of complex lipids desorbed directly from lyophilised membranes and cells. Biochem. Biophys. Res. Com. 142: 194–199.

    Article  CAS  Google Scholar 

  5. Matsuo, T., Caprioli, R.M., and Gross, M.L., (eds.). 1994. Biological mass spectrometry. John Wiley and Sons, Chichester, UK.

    Google Scholar 

  6. Hillenkamp, F., Karas, M., Beavis, R.C., and Chait, B.T. 1991. Matrix assisted laser desorption ionization mass spectrometry of biopolymers. Anal. Chem. 63: 1193A–1202A.

    Article  CAS  Google Scholar 

  7. Cotter, R.J. 1992. Time of flight mass spectrometry for the structural analysis of biological molecules. Anal. Chem. 64: 1027A–1039A.

    Article  CAS  Google Scholar 

  8. Liang, X., Zheng, K., Qian, M.G., and Lubman, D.M. 1996. Determination of bacterial protein profiles by matrix assisted laser desorption ionization mass spectrometry with high performance liquid chromatography. Rapid Commun. Mass Spectrom. 10: 1219–1226.

    Article  CAS  Google Scholar 

  9. Krishnamurthy, T., Ross, P.L., and Rajamani, U. 1996. Detection of pathogenic and non-pathogenic bacteria by matrix assisted laser desorption ionization time of flight mass spectrometry. Rapid Commun. Mass Spectrom. 10: 883–888.

    Article  CAS  Google Scholar 

  10. Holland, R.D., Wilkes, J.G., Rafii, F., Sutherland, J.B., Persons, C.C., Voorhees, K.J. et al. 1996. Rapid identification of intact whole bacteria based on spectral patterns using matrix assisted laser desorption ionization with time of flight mass spectrometry. Rapid Commun. Mass Spectrom. 10: 1227–1232.

    Article  CAS  Google Scholar 

  11. Ewing, W.H. 1986. Edwards and Ewings identification of enterobacteriacae, 4th ed., Elsevier, Amsterdam.

    Google Scholar 

  12. Zubay, G. 1993. Biochemistry, 3rd ed., pp. 138–160. Wm. C. Brown, Dubuque, IA.

    Google Scholar 

  13. Butler, W.R., Jost, K.C. Jr., and Kilburn, J.O. 1991. Identification of mycobacteria by high performance liquid chromatography. J. Clin. Microbiol. 29: 2468–2472.

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Hagen, S.R. and Thompson, J.D. 1995. Analysis of mycolic acids by high performance liquid chromatography and fluorimetric detection—applications for the identification of mycobacteria in clinical samples. J. Chromatog. A. 693: 167–172.

    Article  Google Scholar 

  15. Dobson, G., Minnikin, D.E., Minnikin, S.M., Parlett, J.H., Goodfellow, M., Ridell, M. 1985. Systematic analysis of complex mycobacterial liquids, pp. 237–265 in Chemical Methods in Bacterial Systematics, Society of Applied Bacteriology, UK.

    Google Scholar 

  16. Brennan, P.J. and Nikaido, H. 1995. The envelope of mycobacteria. Ann. Rev. Biochem. 64: 29–63.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Claydon, M., Davey, S., Edwards-Jones, V. et al. The rapid identification of intact microorganisms using mass spectrometry. Nat Biotechnol 14, 1584–1586 (1996). https://doi.org/10.1038/nbt1196-1584

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt1196-1584

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing