Abstract
Advances in MALDI-TOF mass spectrometry have enabled the development of a rapid, accurate and specific method for the identification of bacteria directly from colonies picked from culture plates, which we have named the MALDI Biotyper. The picked colonies are placed on a target plate, a drop of matrix solution is added, and a pattern of protein molecular weights and intensities, “the protein fingerprint” of the bacteria, is produced by the MALDI-TOF mass spectrometer. The obtained protein mass fingerprint representing a molecular signature of the microorganism is then matched against a database containing a library of previously measured protein mass fingerprints, and scores for the match to every library entry are produced. An ID is obtained if a score is returned over a pre-set threshold. The sensitivity of the techniques is such that only approximately 104 bacterial cells are needed, meaning that an overnight culture is sufficient, and the results are obtained in minutes after culture. The improvement in time to result over biochemical methods, and the capability to perform a non-targeted identification of bacteria and spores, potentially makes this method suitable for use in the detect-to-treat timeframe in a bioterrorism event. In the case of white-powder samples, the infectious spore is present in sufficient quantity in the powder so that the MALDI Biotyper result can be obtained directly from the white powder, without the need for culture. While spores produce very different patterns from the vegetative colonies of the corresponding bacteria, this problem is overcome by simply including protein fingerprints of the spores in the library. Results on spores can be returned within minutes, making the method suitable for use in the “detect-to-protect” timeframe.
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References
Demirev PA, Feneslau C (2008) Mass spectrometry for rapid characterization of microorganisms. Ann Rev Anal Chem 1:71–93 and Fenselau C, Demirev P (2001) Characterization of intact microorganisms by MALDI mass spectrometry. Mass Spectrom Rev 20:157–171
Demirev PA, Fenselau C (2008) Mass spectrometry in biodefense. J Mass Spectrom 43:1441–1457
Hathout Y, Demirev PA, Ho Y-P, Bundy JL, Ryzhov V, Sapp L, Stutler J, Jackman J, Fenselau C (1999) Identification of Bacillus spores by matrix-assisted laser desorption ionization-mass spectrometry. Appl Environ Microbiol 65(10):4313–4319
Anhalt JP, Fenselau C (1975) Identification of bacteria using mass spectrometry. Anal Chem 47:219–225
Eigner U, Holfelder M, Oberdorfer K, Betz-Wild U, Bertsch D, Fahr AM (2009) Performance of a matrix-assisted laser desorption ionization-time-of-flight mass spectrometry system for the identification of bacterial isolates in the clinical routine laboratory. Clin Lab 55(7–8):289–296
Seng P, Drancourt M, Gouriet F, La Scola B, Fournier PE, Rolain JM, Raoult D (2009) Ongoing revolution in bacteriology: routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clin Infect Dis 49(4):543–551
Lay JO Jr (2001) MALDI-TOF mass spectrometry of bacteria. Mass Spectrom Rev 20(4):172–194
Krishnamurthy T, Ross PL, 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
Ryzhov V, Feneslau C (2001) Characterization of the protein subset desorbed by MALDI from whole bacterial cells. Anal Chem 73:746–750
Ilina EN, Borovskaya AD, Malakhova MM, Vereshchagin VA, Kubanova AA, Kruglov AN, Svistunova TS, Gazarian AO, Maier T, Kostrzewa M, Govorun VM (2009) Direct bacterial profiling by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry for identification of pathogenic Neisseria. J Mol Diagn 11(1):75–86
Mellmann A, Cloud J, Maier T, Keckevoet U, Ramminger I, Iwen P, Dunn J, Hall G, Wilson D, Lasala P, Kostrzewa M, Harmsen D (2008) Evaluation of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in comparison to 16S rRNA gene sequencing for species identification of nonfermenting bacteria. J Clin Microbiol 46:1946–1954
Mellmann A, Bimet F, Bizet C, Borovskaya AD, Drake RR, Eigner U, Fahr AM, He Y, Ilina EN, Kostrzewa M, Maier T, Mancinelli L, Moussaoui W, Prévost G, Putignani L, Seachord CL, Tang YW, Harmsen D (2009) High interlaboratory reproducibility of matrix-assisted laser desorption ionization-time of flight mass spectrometry-based species identification of nonfermenting bacteria. J Clin Microbiol 47:3732–3734
Grosse-Herrenthey A, Maier T, Gessler F, Schaumann R, Böhnel H, Kostrzewa M, Krüger M (2008) Challenging the problem of Clostridial identification with matrix-assisted laser desorption/ionisation – time of flight mass spectrometry (MALDI – TOF MS). Anaerobe 14(4):242–249
van Veen SQ, Claas ECJ, Kuijper EJ (2010) High-throughput identification of bacteria and yeast by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF MS) in routine medical microbiology laboratory. J Clin Microbiol 48(3):900–907
Cherkaoui A, Hibbs J, Emonet S, Tangomo M, Girard M, Francois P, Schrenzel J (2010) Comparison of two matrix-assisted laser desorption ionization-time of flight mass spectrometry methods with conventional phenotypic identification for routine bacterial speciation. J Clin Microbiol 48(4):1169–1175
La Scola B, Raoult D (2009) Direct identification of bacteria in positive blood culture bottles by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry. PLoS One 25:e8041
Stevenson LG, Drake SK, Murray PR (2010) Rapid identification of bacteria in positive blood culture broths by MALDI-TOF mass spectrometry. J Clin Microbiol 48(2):444–447
Russel AD (1990) Bacterial spores and chemical sporicidal agents. Clin Microbiol Rev 3(2):99–119
Henderson I, Duggleby CJ, Turnbull PC (1994) Differentiation of Bacillus anthracis from other Bacillus cereus group bacteria with the PCR. Int J Syst Bacteriol 44:99–105
Hathout Y, Setlow B, Cabrera-Martinez RM, Fenselau C, Setlow P (2003) Small, acid-soluble proteins as biomarkers in mass spectrometry analysis of Bacillus spores. Appl Environ Microbiol 69(2):1100–1107
Lasch P, Nattermann H, Erhard M, Stämmler M, Grunow R, Bannert N, Appel B, Naumann D (2008) MALDI-TOF mass spectrometry compatible inactivation method for highly pathogenic microbial cells and spores. Anal Chem 80:2026–2034
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Elssner, T., Kostrzewa, M., Maier, T., Kruppa, G. (2011). Microorganism Identification Based On MALDI-TOF-MS Fingerprints. In: Banoub, J. (eds) Detection of Biological Agents for the Prevention of Bioterrorism. NATO Science for Peace and Security Series A: Chemistry and Biology. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9815-3_7
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DOI: https://doi.org/10.1007/978-90-481-9815-3_7
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