Abstract

DNA coding for the 16S rRNA of six strains of the obligate intracellular bacterium Coxiella burnetti was directly amplified from lysed host cells using the polymerase chain reaction. The amplification product was sequenced using a linear-PCR procedure and compared with other published 16S rRNA sequences. The results of this analysis confirm the position of C. burnetii in the gamma subgroup of the proteobacteria. The data show that all of the C. burnetii strains are highly related (> 99%) on the basis of 16S rRNA sequences although they had different geographic origins and phenotypic characteristics. The data support a phylogenetic homogeneity of the genus Coxiella with only one species which is C. burnetii.

References

1
Marrie
T.J.
(
1990
)
Coxiella burnetii (Q fever)
In
Principles and practice of infectious diseases
 
Mandell
G.L.
,
Douglas
R.G.J.
Bennett
J.E.
, Eds)
3rd edn.
, pp
1472
1476
Churchill Livingstone
,
New York
.
2
Weiss
E.
Moulder
J.W.
(
1984
)
Order Ricettsiales, Gieszczkiewicz 1939
In
Bergey's Manual of Systematic Bacteriology
 
Kreig
N.R.
Holt
J.G.
, Eds) Vol.
1
, pp
687
701
Williams and Williams
,
Baltimore, MD
.
3
Stein
A.
Raoult
D.
(
1992
)
Phenotypic and genotypic heterogeneity of 8 new human Coxiella burnetii isolates
Acta. Virol. (Praha)
 ,
36
,
7
12
.
4
Hendrix
L.R.
Samuel
J.E.
Mallavia
L.P.
(
1991
)
Differenciation of Coxiella burnetii isolates by analysis of restriction-endonuclease-digested DNA separated by SDS-PAGE
J. Gen. Microbiol.
 ,
137
,
269
276
.
5
Weisburg
W.G.
Dobson
M.E.
Samuel
J.E.
Dasch
G.A.
Mallavia
L.P.
Baca
O.
Mandelco
L.
Sechrest
J.E.
Weiss
E.
Woese
C.R.
(
1989
)
Phylogenetic diversity of the Rickettsiae
J. Bacteriol.
 ,
171
,
4202
4206
.
6
Raoult
D.
Vestris
G.
Enea
M.
(
1990
)
Isolation of 16 strains of Coxiella burnetii from patients by using a sensitive centrifugation cell culture system and establishment of strains in HEL cells
J. Clin. Microbiol.
 ,
28
,
2482
2484
.
7
Stein
A.
Raoult
D.
(
1992
)
Rapid detection of Coxiella burnetii in paraffin - embedded tissues using Chelex R 100 and PCR
Nucl. Acid. Res.
 ,
20
,
5237
5238
.
8
Embley
T.M.
(
1991
)
The linear PCR reaction: a simple and robust method for sequencing amplified rRNA genes
Lett. Appl. Microbiol.
 ,
13
,
171
174
.
9
Higgins
D.G.
Sharp
P.M.
(
1989
)
Fast and sensitive multiple sequence alignments on a microcomputer
Cabios
 ,
5
,
151
153
.
10
Felsenstein
J.
(
1989
)
PHYLIP-Phylogeny inference package
Cladistics
 ,
5
,
164
166
.
11
Jukes
T.H.
Cantor
C.R.
(
1969
) In
Mammalian protein metabolism
 
Evolution of protein molecules
 .
Munro
H.N.
, Ed) pp
121
132
.
12
Fitch
W.M.
Margoliash
E.
(
1967
)
Construction of phylogenetic trees: a method based on mutational distances as estimated from cytochrome c sequences is of general applicability
Science
 ,
155
,
279
284
.
13
Vodkin
M.H.
Williams
J.C.
Stephenson
E.H.
(
1986
)
Genetic heterogeneity among isolates of Coxiella burnetii
J. Gen. Microbiol.
 ,
132
,
455
463
.
14
Savinelli
E.A.
Mallavia
L.P.
(
1990
)
Comparison of Coxiella burnetii plasmids to homologous chromosomal sequences present in a plasmidless endocarditis-causing isolate
Ann. NY. Acad. Sci.
 ,
590
,
523
533
.
15
Heinzen
R.
Stiegler
G.L.
Whiting
L.L.
Schmitt
S.A.
Mallavia
L.P.
Frazier
M.E.
(
1990
)
Use of pulsed field gel electrophoresis to differentiate Coxiella burnetii strains
Ann. NY. Acad. Sci.
 ,
590
,
504
513
.