Rapid and Culture Free Identification of Francisella in Hare Carcasses by High-Resolution Tandem Mass Spectrometry Proteotyping

Natalie Witt; Sandro Andreotti; Anne Busch; Kerstin Neubert; Knut Reinert; Herbert Tomaso; David Meierhofer

doi:10.3389/fmicb.2020.00636

Rapid and Culture Free Identification of Francisella in Hare Carcasses by High-Resolution Tandem Mass Spectrometry Proteotyping

Front Microbiol. 2020 May 8:11:636. doi: 10.3389/fmicb.2020.00636. eCollection 2020.

Authors

Natalie Witt^{1

2}, Sandro Andreotti³, Anne Busch^{4

5}, Kerstin Neubert⁶, Knut Reinert⁶, Herbert Tomaso⁴, David Meierhofer¹

Affiliations

¹ Mass Spectrometry Facility, Max Planck Institute for Molecular Genetics, Berlin, Germany.
² Bioanalytics, Institute of Biotechnology, Technische Universität Berlin, Berlin, Germany.
³ Bioinformatics Solution Center, Department of Mathematics and Computer Science, Freie Universität Berlin, Germany.
⁴ Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, Jena, Germany.
⁵ Universitätsklinikum Jena, Friedrich-Schiller-Universität, Jena, Germany.
⁶ Department of Mathematics and Computer Science, Freie Universität Berlin, Berlin, Germany.

Abstract

Zoonotic pathogens that can be transmitted via food to humans have a high potential for large-scale emergencies, comprising severe effects on public health, critical infrastructures, and the economy. In this context, the development of laboratory methods to rapidly detect zoonotic bacteria in the food supply chain, including high-resolution mass spectrometry proteotyping are needed. In this work, an optimized sample preparation method for liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteome profiling was established for Francisella isolates, and a cluster analysis, as well as a phylogenetic tree, was generated to shed light on evolutionary relationships. Furthermore, this method was applied to tissues of infected hare carcasses from Germany. Even though the non-informative data outnumbered by a manifold the information of the zoonotic pathogen in the resulting proteome profiles, the standardized evaluation of MS data within an established automated analysis pipeline identified Francisella (F.) tularensis and, thus, could be, in principle, an applicable method to monitor food supply chains.

Keywords: Francisella tularensis; detection of pathogens; high-resolution tandem mass spectrometry proteotyping; phylogenetic analysis; proteome profiling.