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PLoS One. 2015 Apr 9;10(4):e0123298. doi: 10.1371/journal.pone.0123298. eCollection 2015.

Rapid high resolution genotyping of Francisella tularensis by whole genome sequence comparison of annotated genes ("MLST+").

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Bundeswehr, Institute of Microbiology, Neuherbergstrasse 12, 80937, Muenchen, Germany.
University of Muenster, Department of Periodontology, Waldeyerstrasse 30, 48149, Muenster, Germany.
University of Muenster, Institute of Hygiene, Robert-Koch-Strasse 41, 48149, Muenster, Germany.
Bundeswehr, Institute of Microbiology, Neuherbergstrasse 12, 80937, Muenchen, Germany; Rostock University Hospital, Institute of Medical Microbiology, Virology and Hygiene, Schillingallee 70, 18057 Rostock, Germany.


The zoonotic disease tularemia is caused by the bacterium Francisella tularensis. This pathogen is considered as a category A select agent with potential to be misused in bioterrorism. Molecular typing based on DNA-sequence like canSNP-typing or MLVA has become the accepted standard for this organism. Due to the organism's highly clonal nature, the current typing methods have reached their limit of discrimination for classifying closely related subpopulations within the subspecies F. tularensis ssp. holarctica. We introduce a new gene-by-gene approach, MLST+, based on whole genome data of 15 sequenced F. tularensis ssp. holarctica strains and apply this approach to investigate an epidemic of lethal tularemia among non-human primates in two animal facilities in Germany. Due to the high resolution of MLST+ we are able to demonstrate that three independent clones of this highly infectious pathogen were responsible for these spatially and temporally restricted outbreaks.

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