Abstract
Mass spectrometry (MS) has been widely used in recent years for bacterial identification and typing. Single bacterial colonies are regarded as pure cultures of bacteria grown from single cells. In this chapter, we describe a method for identifying bacteria at the species level with 100% accuracy using the proteomes of bacterial cultures from single colonies. In this chapter, six reference strains of gram-negative and gram-positive bacteria are analyzed, producing results of high reproducibility, as examples of bacterial identification through the application of liquid chromatography–tandem mass spectrometry (LC-MS/MS) and a custom database. Details on sample preparation and identification of Streptococcus pneumoniae are also described.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Cheng K, Chui H, Domish L et al (2016) Recent development of mass spectrometry and proteomics applications in identification and typing of bacteria. Proteomics Clin Appl 10:346–357. https://doi.org/10.1002/prca.201500086
Cheng K, Sloan A, Peterson L, McCorrister S et al (2014) Comparative study of traditional flagellum serotyping and liquid chromatography-tandem mass spectrometry-based flagellum typing with clinical Escherichia coli isolates. J Clin Microbiol 52(6):2275–2278. https://doi.org/10.1128/JCM.00174-14
Chui H, Chan M, Hernandez D et al (2015) Rapid, sensitive, and specific Escherichia coli H antigen typing by matrix-assisted laser desorption ionization-time of flight-based peptide mass fingerprinting. J Clin Microbiol 53(8):2480–2485. https://doi.org/10.1128/JCM.00593-15
Cheng K, She YM, Chui H et al (2016) Mass spectrometry-based Escherichia coli H antigen/flagella typing: validation and comparison with traditional serotyping. Clin Chem 62(6):839–847. https://doi.org/10.1373/clinchem.2015.244236
Cheng K, Sloan A, McCorrister S et al (2014) Quality evaluation of LC-MS/MS-based E. coli H antigen typing (MS-H) through label-free quantitative data analysis in a clinical sample setup. Proteomics Clin Appl 8:963–970. https://doi.org/10.1002/prca.201400019
Ishihama Y, Oda Y, Tabata T et al (2005) Exponentially modified protein abundance index (emPAI) for estimationof absolute protein amount in proteomics by the number of sequencedpeptides per protein. Mol Cell Proteomics 4(9):1265–1272. https://doi.org/10.1074/mcp.M500061-MCP200
Tracz DM, McCorrister SJ, Chong PM et al (2013) A simple shotgun proteomics method for rapid bacterial identification. J Microbiol Methods 94(1):54–57. https://doi.org/10.1016/j.mimet.2013.04.008
Acknowledgments
We thank Stuart McCorrister and Chris Grant in the Proteomics Core, National Microbiology Laboratory, Public Health Agency of Canada for creating the database and running the MS.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Zhou, J., Zhang, L., Chuan, H., Sloan, A., Tsang, R., Cheng, K. (2019). Mass Spectrometry to Study the Bacterial Proteome from a Single Colony. In: Iovino, F. (eds) Streptococcus pneumoniae. Methods in Molecular Biology, vol 1968. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9199-0_10
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9199-0_10
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9198-3
Online ISBN: 978-1-4939-9199-0
eBook Packages: Springer Protocols