Assessing the ratio of Bacillus spores and vegetative cells by shotgun proteomics
Mass spectrometry for rapid identification of microorganisms is expanding over the last years because this approach is quick. This methodology provides a decisive interest to fight against bioterrorism as it is applicable whatever the pathogen to be considered and often allows subtyping which may be crucial for confirming a massive and widespread attack with biological agents. Here, we present a methodology based on next-generation proteomics and tandem mass spectrometry for discovering numerous protein biomarkers allowing the discrimination of spores and vegetative cells of Bacillus atrophaeus, a biowarfare simulant. We propose a global quantitative evaluation of the two groups of discriminant biomarkers based on their aggregated normalized spectral abundance factors.
KeywordsBacillus Detection Mass spectrometry Shotgun proteomics Spores
We thank Jean-Charles Gaillard and Guylaine Miotello for invaluable technical help with the mass spectrometry platform.
This work was funded in part by the French joint ministerial program of R&D against CBRNE threats and the ANR program “Phylopeptidomics” (ANR-17-CE18-0023-01). CM was supported by DGA and CEA
- Carvalho PC, Fischer JS, Xu T, Yates JR 3rd, Barbosa VC (2012) PatternLab: from mass spectra to label-free differential shotgun proteomics. Curr Protoc Bioinformatics Chapter 13:Unit13.19Google Scholar
- Eng C, Blouin Y, Ding N, Larigauderie G, Ramisse V, Pujol C (2015) Draft genome sequence of the biowarfare simulant Bacillus atrophaeus strain 930029. Genome Announc 3Google Scholar
- Raymond B, Federici BA (2017) In defense of Bacillus thuringiensis, the safest and most successful microbial insecticide available to humanity—a response to EFSA. FEMS Microbiol Ecol 93Google Scholar
- Rougemont B, Bontemps Gallo S, Ayciriex S, Carriere R, Hondermarck H, Lacroix JM, Le Blanc JC, Lemoine J (2017) Scout-MRM: multiplexed targeted mass spectrometry-based assay without retention time scheduling exemplified by Dickeya dadantii proteomic analysis during plant infection. Anal Chem 89:1421–1426CrossRefGoogle Scholar
- Srivastava A, Pitesky ME, Steele PT, Tobias HJ, Fergenson DP, Horn JM, Russell SC, Czerwieniec GA, Lebrilla CB, Gard EE, Frank M (2005) Comprehensive assignment of mass spectral signatures from individual Bacillus atrophaeus spores in matrix-free laser desorption/ionization bioaerosol mass spectrometry. Anal Chem 77:3315–3323CrossRefGoogle Scholar