title: Identification and subtyping of Francisella by pyrosequencing and signature matching of 16S rDNA fragments.
abstract: OBJECTIVE
  To analyse the V1 region of the 16S rDNA gene by a universal pyrosequencing protocol to identify and subtype Francisella in 31 strains from a repository collection and 96 patient isolates.
  
  
  RESULTS
  Pyrosequencing was used to determine the nucleotide sequence of PCR amplification products of the variable region (V1) of the 16S rDNA from 31 repository strains and 96 isolates from Swedish patients with ulceroglandular tularaemia. Pyrosequencing resulted in a 37 nucleotide sequence, specific for Francisella sp., for all repository strains and patient samples analysed. In addition, the isolates could be divided into two groups based on the analysis of a single nucleotide polymorphism in the sequence: one group included Francisella tularensis ssp. tularensis, ssp. holarctica and ssp. mediasiatica, whereas the other group included Francisella tularensis ssp. novicida and other species of Francisella. The analysis of samples taken from patients suffering from ulceroglandular tularaemia revealed that all isolates belonged to the first group comprising subspecies of F. tularensis virulent for humans.
  
  
  CONCLUSIONS
  The pyrosequencing analysis of the 16S rDNA V1 is a useful molecular tool for the rapid identification of suspected isolates of Francisella sp. in clinical or environmental samples.
  
  
  CONCLUSIONS
  Virulent F. tularensis ssp. causing ulceroglandular tularaemia, or those with a potential to be used in a bioterrorism event, could rapidly be discriminated from subspecies less virulent for humans.
authors.affiliation: Center for Biological Security (ZBS 2), Robert Koch-Institut, Berlin, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Jacob
authors.forename: T
authors.initials: T
authors.lastname: Wahab
authors.forename: B
authors.initials: B
authors.lastname: Edvinsson
authors.forename: A
authors.initials: A
authors.lastname: Peterzon
authors.forename: T
authors.initials: T
authors.lastname: Boskani
authors.forename: L
authors.initials: L
authors.lastname: Farhadi
authors.forename: A
authors.initials: A
authors.lastname: Barduhn
authors.forename: R
authors.initials: R
authors.lastname: Grunow
authors.forename: G
authors.initials: G
authors.lastname: Sandström
mesh_terms: D001483:Base Sequence; D004275:DNA, Ribosomal / Q000235:genetics; D005260:Female; D005603:Francisella / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D005604:Francisella tularensis / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D006801:Humans; D008297:Male; D058889:Molecular Typing / Q000379:methods*; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D014406:Tularemia / Q000382:microbiology*
chemical_list: D004275:DNA, Ribosomal; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S