Abstract
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been used for many years to rapidly identify whole bacteria. However, no consistent methodology exists for the rapid identification of bacteria cultured in liquid media. Thus, in this study we explored the use of MALDI-TOF MS analysis for rapid identification of cells cultured in liquid media. We determined that 2,5-dihydroxybenzoic acid (50 mg mL−1, 50% acetonitrile, 0.1% trifluoroacetic acid) was the best matrix solution for MALDI-TOF MS for this type of study. Moreover, the tested strains were successfully differentiated by principal component analysis, and the main characteristics of the mass peaks for each species were found in mixed culture samples. In addition, we found that the minimum number of cells for detection was 1.8×103. In conclusion, our findings suggest that MS-based techniques can be developed as an auxiliary method for rapidly and accurately identifying bacteria cultured in liquid media.
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Somtcjemko V, Iredell J R, Gilbert G L. Pathogen profiling for disease management and surveillance. Nat Rev Microbiol, 2007, 5: 464–470, 10.1038/nrmicro1656
Widjojoatmodjo M N, Fluit A C, Verhoef J. Rapid identification of bacteria by PCR-single-strand conformation polymorphism. Clin Microbiol, 1994, 32: 3002–3007, 1:CAS:528:DyaK2MXivFWnu78%3D
Pershing D H, Tenover F C, Versalovic J, et al. Molecular microbiology: Diagnostic principles and practice, 8th ed. Washington DC: American Society for Microbiology Press, 2003
Anhalt J P, Fenselau C. Identification of bacteria using mass spectrometry. Anal Chem, 1975, 47: 219–225, 1:CAS:528:DyaE2MXot1CrsA%3D%3D, 10.1021/ac60352a007
Ryzhov V, Fenselau C. Characterization of the protein subset desorbed by MALDI from whole bacterial cells. Anal Chem, 2001, 73: 746–750, 1:CAS:528:DC%2BD3MXkt1KitA%3D%3D, 10.1021/ac0008791, 11248887
Demirev P A, Ramirez J, Fenselau C. Tandem mass spectrometry of intact proteins for characterization of biomarkers from Bacillus cereus T spores. Anal Chem, 2001, 73: 5725–5731, 1:CAS:528:DC%2BD3MXnslOhs78%3D, 10.1021/ac010672n, 11774913
Warscheid B, Fenselau C. A targeted proteomics approach to the rapid identification of bacterial cell mixtures by matrix-assisted laser desorption/ionization mass spectrometry. Proteomics, 2004, 4: 2877–2892, 1:CAS:528:DC%2BD2cXovVynsbw%3D, 10.1002/pmic.200400911, 15378756
Pribil P A, Patton E, Black G, et al. Rapid characterization of Bacil lus spores targeting species-unique peptides produced with an atmospheric pressure matrix-assisted laser desorption/ionization source. J Mass Spectrom, 2005, 40: 464–474, 1:CAS:528:DC%2BD2MXktFKjtbk%3D, 10.1002/jms.816, 15712356
Sauer S, Freiwald A, Maier T, et al. Classification and identification of bacteria by mass spectrometry and computational analysis. PLoS One, 2008, 3: e2843, 10.1371/journal.pone.0002843, 18665227
Carbonnelle E, Beretti J L, Cottyn S, et al. Rapid identification of Staphylococci isolated in clinical microbiology laboratories by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol, 2007, 45: 2156–2161, 1:CAS:528:DC%2BD2sXptFGltr4%3D, 10.1128/JCM.02405-06, 17507519
Mellmann A, Cloud J, Maier T, et al. Evaluation of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry in comparison to 16S rRNA gene sequencing for species identification of nonfermenting bacteria. J Clin Microbiol, 2008, 46: 1946–1954, 1:CAS:528:DC%2BD1cXotFGhs7c%3D, 10.1128/JCM.00157-08, 18400920
Piseth S, Michel D, Frederique G, et al. Ongoing revolution in bacteriology: Routine identification of bacteria by Matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Clin Infect Dis, 2009, 49: 543–551, 10.1086/600885
Torsten S. Whole-cell matrix-assisted laser desorption/ionization mass spectrometry for rapid identification of bacteriocin/lantibiotic-producing bacteria. Rapid Commun Mass Spectrom, 2008, 22: 1146–1152, 10.1002/rcm.3481
Hoben H J, Somasegaran P. Comparison of the pour, spread and drop plate methods for enumeration of Rhizobium spp. in inoculants made from presterilized peat. Appl Environ Microbial, 1982, 44: 1246–1247, 1:STN:280:DC%2BC3crnvFKltg%3D%3D
Song Y S, Talaty N, Tao W A, et al. Rapid ambient mass spectrometric profiling of intact, untreated bacteria using desorption electrospray ionization. Chem Commun, 2007, 1: 61–63, 10.1039/b615724f
Lindon J C, Holmes E, Nicholson J K. Metabonomics techniques and applications to pharmaceutical research & development. Pharm Res, 2006, 23: 1075–1088, 1:CAS:528:DC%2BD28XmtFCnsL0%3D, 10.1007/s11095-006-0025-z, 16715371
Hsieh S Y, Tseng C L, Lee Y S, et al. Highly efficient classification and identification of human pathogenic bacteria by MALDI-TOF MS. Mol Cell Proteomics, 2008, 7: 448–456, 1:CAS:528:DC%2BD1cXitlylt70%3D, 18045801
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Zhou, N., Wang, N., Xu, B. et al. Whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for rapid identification of bacteria cultured in liquid media. Sci. China Life Sci. 54, 48–53 (2011). https://doi.org/10.1007/s11427-010-4119-9
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DOI: https://doi.org/10.1007/s11427-010-4119-9