Direct Bacterial Profiling by Matrix-Assisted Laser Desorption−Ionization Time-of-Flight Mass Spectrometry for Identification of Pathogenic Neisseria

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Direct Bacterial Profiling by Matrix-Assisted Laser Desorption−Ionization Time-of-Flight Mass Spectrometry for Identification of Pathogenic Neisseria Elena N. Ilina, Alexandra D. Borovskaya, Maja M. Malakhova, Vladimir A. Vereshchagin, Anna A. Kubanova, Alexander N. Kruglov, Tatyana S. Svistunova, Anaida O. Gazarian, Thomas Maier, Markus Kostrzewa, Vadim M. Govorun The Journal of Molecular Diagnostics Volume 11, Issue 1, Pages 75-86 (January 2009) DOI: 10.2353/jmoldx.2009.080079 Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 MALDI-TOF mass spectrum of a whole-cell extract of E. coli strain DH5α. The mass-spectrum was recorded within the range of m/z 2000 to 20,000, zoomed region (m/z 2000 to 4000) illustrates peaks of doubly charged ions. The roman numerals indicate signal pairs of singly and doubly charged ions of the same molecules. The relative intensities of the ions are shown on the y axis, the mass to charge ratios are shown on the x axis. The Journal of Molecular Diagnostics 2009 11, 75-86DOI: (10.2353/jmoldx.2009.080079) Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 MALDI-TOF mass spectra of whole-cell extracts of pathogenic Neisseria strains: N. gonorrhoeae strain ATCC 49226 (A) and N. meningitidis clinical strain 537 (B). The relative intensities of the ions are shown on the ϒ axis, the mass to charge ratios on the x axis. Changes of MALDI-TOF mass spectra observed for clinical strains of N. gonorrhoeae (C) and N. meningitidis (D). Variants of peaks corresponding to ribosomal proteins RL 36, RL 34, and RL 31 of N. gonorrhoeae and of those corresponding to ribosomal proteins RS 20 and RL27 of N. meningitidis are shown. The Journal of Molecular Diagnostics 2009 11, 75-86DOI: (10.2353/jmoldx.2009.080079) Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 MALDI-TOF mass spectra of the purified ribosomal fraction and cell lysate of N. gonorrhoeae ATCC 49226 strain. The relative intensities of the ions are shown on the y axis, and the mass to charge ratios are shown on the x axis. The Journal of Molecular Diagnostics 2009 11, 75-86DOI: (10.2353/jmoldx.2009.080079) Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 MALDI-TOF mass spectra comparison of closely related species of N. gonorrhoeae, N. meningitidis, N. sicca, N. preflava, and non-determined Neisseria spp 8503. The relative intensities of the ions are shown on the ϒ axis, and the mass to charge ratios are shown on the x axis. The patterns display the individual distinguishes among isolates that lead to Neisseria species separation. The Journal of Molecular Diagnostics 2009 11, 75-86DOI: (10.2353/jmoldx.2009.080079) Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 5 A score oriented dendrogram of MALDI-TOF mass spectra profiles was generated using the MALDI Biotyper 1.1 software with the following settings for peak picking: lower bound 3000, upper bound 11,000, maximum peaks 100 and threshold 0.001. Neisseria species are listed in Table 1. The dendrogram was generated with the following settings: the distance measure was set at Euclidian and linkage set at completion. The Journal of Molecular Diagnostics 2009 11, 75-86DOI: (10.2353/jmoldx.2009.080079) Copyright © 2009 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions