Analysis of 16S Ribosomal DNA Sequences of Francisella Strains and Utilization for Determination of the Phylogeny of the Genus and for Identification of Strains by PCR
The 16S ribosomal DNAs (rDNAs) of two strains of Francisella tularensis and one strain of Francisella philomiragia were sequenced. On the basis of phylogenetic analysis data, the genus Francisella was placed in the ? subclass of the Proteobacteria. The most closely related organism was the intracellular bacterium Wolbachia persica. The sequenced 16S rDNA molecules of the Francisella species exhibited very high levels of similarity (98.5 to 99.9%). Two variable regions, comprising 390 to 450 nucleotides of the 16S rDNA molecules of 17 additional Francisella strains, including members of the species F. tularensis and F. philomiragia, were also sequenced. At most, six nucleotide differences were observed among the sequences of the F. tularensis strains. The sequence of Francisella novicida was virtually identical to the sequences of the F. tularensis strains, thereby supporting the hypothesis that these organisms are members of the same species. On the basis of the observed differences, primer pairs were designed to distinguish strains by using the PCR at the genus, species, and subspecies levels. This permitted sensitive identification of strains belonging to the genus Francisella and discrimination of the species F. tularensis and F. philomiragia.
DamsE.,
HendriksL.,
Van de PeerY.,
NeefsJ.-M.,
SmitsG.,
VandenbemptI.,
de WächterR.1988;
Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Res. Suppl.16v87v173
DewhirstF. E.,
PasterB. J.,
OlsenI.,
FraserG. J.1992;
Phylogeny of 54 representative strains of species in the family Pasteurellaceae as determined by comparison of 16S rRNA sequences. J. Bacteriol.17420022013
EigelsbachH. T.,
DownsC. M.1961;
Prophylactic effectiveness of live and killed tularemia vaccines. I. Production of vaccine and evaluation in the white mouse and guinea pig. J. Immunol.87415425
FitchW. M.,
MargoliashE.1967;
Construction of phylogenetic trees: a method based on mutational distances as estimated from cytochrome c sequences is of general applicability. Science155279284
ForemanM.,
SandströmG.,
JaurinB.1990;
Identification of Francisella species and discrimination of type A and type B strains of F. tularensis by 16S rRNA analysis. Appl. Environ. Microbiol.56949955
FoxG. E.,
WisotzkeyJ. D.,
JurtshukP.Jr.1992;
How close is close: 16S rRNA sequence identity may not be sufficient to guarantee species identity. Int. J. Syst. Bacteriol.42166170
FryN. K.,
WarwickS.,
SaundersN. A.,
EmbleyT. M.1991;
The use of 16S ribosomal RNA analyses to investigate the phylogeny of the family Legionellaceae. J. Gen. Microbiol.13712151222
HollisD. G.,
WeaverR. E.,
SteigerwaltA. G.,
WengerJ. D.,
MossC. W.,
BrennerD. J.1989;
Francisella philomiragia comb. nov. (formerly Yersinia philomiragia) and Francisella tularensis biogroup novicida (formerly Francisella novicida) associated with human disease. J. Clin. Microbiol.2716011608
HultmanT.,
StåhlS.,
HornesE.,
UhlénM.1989;
Direct solid phase sequencing of genomic and plasmid DNA using magnetic beads as a support. Nucleic Acids Res.1749374946
KimuraM.1980;
A simple model for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J. Mol. Evol.16111120
MarconiR. T.,
GaronC. F.1992;
Phylogenetic analysis of the genus Borrelia: a comparison of North American and European isolates of Borrelia burgdorferi. J. Bacteriol.174241244
Martinez-MurciaA. J.,
BenlochS.,
CollinsD.1992;
Phylogenetic interrelationships of members of the genera Aeromonas and Plesiomonas as determined by 16S ribosomal DNA sequencing: lack of congruence with results of DNA-DNA hybridizations. Int. J. Syst. Bacteriol.42412421
MoodyM. D.,
DownsC. M.1955;
Studies on tularemia. I. The relation between certain pathogenic and immunogenic properties of variants of Pasteurella tularensis. J. Bacteriol.70297304
MurrayR. G. E.,
BrennerD. J.,
ColwellR. R.,
de VosP.,
GoodfellowM.,
GrimontP. A. D.,
PfennigN.,
StackebrandtE.,
ZavarzinG. A.1990;
Report of the Ad Hoc Committee on Approaches to Taxonomy within the Proteobacteria. Int. J. Syst. Bacteriol.40213215
OlsufievN. G.,
EmelyanovaO. S.,
DunayevaT. N.1959;
Comparative study of strains of Bacterium tularense. II. Evaluation of criteria of virulence of Bacterium tularense in the old and the new world and their taxonomy. J. Hyg. Epidemiol. Microbiol. Immunobiol.3138149
SandströmG.,
SjöstedtA.,
ForemanM.,
PavlovichN. V.,
MishankinB. N.1992;
Characterization and classification of strains of Francisella tularensis isolated in the Central Asian focus of the Soviet Union and in Japan. J. Clin. Microbiol.30172175
StackebrandtE.,
MurrayR. G. E.,
TrüperH. G.1988;
Proteobacteria classis nov., a name for the phylogenetic taxon that includes the “purple bacteria and their relatives.” Int. J. Syst. Bacteriol.38321325
WeisburgW. G.,
DobsonM. E.,
SamuelJ. E.,
DaschG. A.,
MallaviaL. P.,
BacaO.,
MandelcoL.,
SechrestJ. E.,
WeissE.,
WoeseC. R.1989;
Phylogenetic diversity of the rickettsiae. J. Bacteriol.17142024206
WeissE.,
DaschG. A.,
ChangK.-P.1984;
Gram-negative aerobic cocci. 711713KriegN. R.,
HoltJ. G.Bergey’s manual of systemic bacteriology1
Williams & Wilkins Co.;
Baltimore:
Analysis of 16S Ribosomal DNA Sequences of Francisella Strains and Utilization for Determination of the Phylogeny of the Genus and for Identification of Strains by PCR