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. 2010 Nov 12:10:285.
doi: 10.1186/1471-2180-10-285.

Rapid identification and typing of Yersinia pestis and other Yersinia species by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry

Saravanan Ayyadurai  1 , Christophe Flaudrops, Didier Raoult, Michel Drancourt
Affiliations

Rapid identification and typing of Yersinia pestis and other Yersinia species by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry

Saravanan Ayyadurai et al. BMC Microbiol. .

Abstract

Background: Accurate identification is necessary to discriminate harmless environmental Yersinia species from the food-borne pathogens Yersinia enterocolitica and Yersinia pseudotuberculosis and from the group A bioterrorism plague agent Yersinia pestis. In order to circumvent the limitations of current phenotypic and PCR-based identification methods, we aimed to assess the usefulness of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) protein profiling for accurate and rapid identification of Yersinia species. As a first step, we built a database of 39 different Yersinia strains representing 12 different Yersinia species, including 13 Y. pestis isolates representative of the Antiqua, Medievalis and Orientalis biotypes. The organisms were deposited on the MALDI-TOF plate after appropriate ethanol-based inactivation, and a protein profile was obtained within 6 minutes for each of the Yersinia species.

Results: When compared with a 3,025-profile database, every Yersinia species yielded a unique protein profile and was unambiguously identified. In the second step of analysis, environmental and clinical isolates of Y. pestis (n = 2) and Y. enterocolitica (n = 11) were compared to the database and correctly identified. In particular, Y. pestis was unambiguously identified at the species level, and MALDI-TOF was able to successfully differentiate the three biotypes.

Conclusion: These data indicate that MALDI-TOF can be used as a rapid and accurate first-line method for the identification of Yersinia isolates.

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Figures

Figure 1
Figure 1
Protein profile of the major Y. pestis biotypes generated by MALDI-TOF-MS. a.i., arbitrary intensity given by the software.
Figure 2
Figure 2
Pseudo-gel representing the protein profile obtained after MALDI-TOF-MS analysis of Y. pestis organisms representative of the Antiqua, Medievalis and Orientalis biotypes. arb.u., arbitrary unit - transcription for arbitrary intensity in the Bruker software; sp# is the numbers of the spectrum.
See this image and copyright information in PMC

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