Francisella tularensis molecular typing using differential insertion sequence amplification

Marilynn A Larson, Paul D Fey, Amanda M. Bartling, Peter Charles Iwen, Michael P. Dempsey, Stephen C. Francesconi, Steven Heye Hinrichs

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Tularemia is a potentially fatal disease that is caused by the highly infectious and zoonotic pathogen Francisella tularensis. Despite the monomorphic nature of sequenced F. tularensis genomes, there is a significant degree of plasticity in the organization of genetic elements. The observed variability in these genomes is due primarily to the transposition of direct repeats and insertion sequence (IS) elements. Since current methods used to genotype F. tularensis are time-consuming and require extensive laboratory resources, IS elements were investigated as a means to subtype this organism. The unique spatial location of specific IS elements provided the basis for the development of a differential IS amplification (DISA) assay to detect and distinguish the more virulent F. tularensis subsp. tularensis (subtypes A.I and A.II) and subsp. holarctica (type B) strains from F. tularensis subsp. novicida and other near neighbors, including Francisella philomiragia and Francisella-like endosymbionts found in ticks. Amplicon sizes and sequences derived from DISA showed heterogeneity within members of the subtype A.I and A.II isolates but not the type B strains. These differences were due to a 312-bp fragment derived from the IS element ISFtu1. Analysis of wild-type F. tularensis isolates by DISA correlated with pulsed-field gel electrophoresis genotyping utilizing two different restriction endonucleases and provided rapid results with minimal sample processing. The applicability of this molecular typing assay for environmental studies was demonstrated by the accurate identification and differentiation of tick-borne F. tularensis. The described approach to IS targeting and amplification provides new capability for epidemiological investigations and characterizations of tularemia source outbreaks.

Original languageEnglish (US)
Pages (from-to)2786-2797
Number of pages12
JournalJournal of clinical microbiology
Volume49
Issue number8
DOIs
StatePublished - Aug 1 2011

Fingerprint

Francisella tularensis
Molecular Typing
Insertional Mutagenesis
DNA Transposable Elements
Francisella
Tularemia
Ticks
Genome
Nucleic Acid Repetitive Sequences
Pulsed Field Gel Electrophoresis
DNA Restriction Enzymes
Zoonoses
Disease Outbreaks
Genotype

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

Francisella tularensis molecular typing using differential insertion sequence amplification. / Larson, Marilynn A; Fey, Paul D; Bartling, Amanda M.; Iwen, Peter Charles; Dempsey, Michael P.; Francesconi, Stephen C.; Hinrichs, Steven Heye.

In: Journal of clinical microbiology, Vol. 49, No. 8, 01.08.2011, p. 2786-2797.

Research output: Contribution to journalArticle

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