The phylogenetic relationships among 36 validly described species or subspecies within the genus Staphylococcus were investigated by cloning an sequencing their 60 kDa heat-shock protein (HSP60) genes using a set of universal degenerate HSP60 PCR primers. The cloned partial HSP60 DNA sequences from nine Staphylococcus aureus strains were highly conserved (97-100% DNA sequence similarity; mean 98%), indicating that the HSP60 gene of multiple isolates within the same species have little microheterogeneity. At the subspecies level, DNA sequence similarity among members of S. aureus, Staphylococcus schleiferi, Staphylococcus cohnii and Staphylococcus capitis ranged from 91 to 98%. At the interspecies level, sequence similarity among 23 distinct species of staphylococci ranged from 7 to 93% (mean 82%). By comparison, the highest sequence similarity of Bacill subtilis and Escherichia coli with members within the genus Staphylococcus was only 70 and 59%, respectively. Importantly, phylogenetic analysis based on the neighbour-joining distance method revealed remarkable concordance between the tree derived from partial HSP60 gene sequences and that based on genomic DNA-DNA hybridization, while 16S rRNA gene sequences correlated less well. The results demonstrate that DNA sequences from the highly conserved and ubiquitous HSP60 gene offer a convenient and accure tool for species-specific identification and phylogenetic analysis of staphylococci.
AltschulS. F.,
MaddenT.,
L, SchafferA. A.,
ZhangJ.,
ZhangZ.,
MillerW.,
LipmanD. J.1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402
BirnbaumD.,
HerwaldtL.,
LowD. E.,
NobleM.,
PfallerM.,
SherertzR.,
ChowA. W.1994; Efficacy of microbial identification system for epidemiologic typing of coagulase- negative staphylococci. J Clin Microbiol 32:2113–2119
EndlJ.,
SeidlH. P.,
FiedlerF.,
SchleiferK. H.1983; Chemical composition and structure of cell wall teichoic acids of staphylococci. Arch Microbiol 135:215–223
FoxG. E.,
WisotzkeyJ. D.,
JurtshukP.Jr1992; How close is close: 16S rRNA sequence identity may not be sufficient to guarantee soecies identity. Int J Syst Bacteriol 42:166–170
GohS. H.,
SantucciZ.,
KloosW. E.,
FaltynM.,
GeorgeC. G.,
DriedgerD.,
HemmingsenS. M.1997; Identification of Staphylococcus species and subspecies by the chaperonin 60 gene identification method and reverse checkerboard hybridization. J Clin Microbiol 35:3116–3121
GribaldoS.,
CooksonB.,
SaundersN.,
MarplesR.,
StanleyJ.1997; Rapid identification by specific PCR of coagulase- negative staphylococcal species important in hospital infection. J Med Microbiol 46:45–53
IevenM.,
VerhoevenJ.,
PattynS. R.,
GoosensH.1995; Rapid and economical method for species identification of clinically significant coagulase-negative staphylococci. J Clin Microbiol 33:1060–1063
JukesT. H.,
CantorC. R.1969; Evolution of protein molecules. In Mammalian Protein Metabolism vol. 3 pp. 21–132 Edited by
MunroH. N.
New York: Academic Press;
KloosW. E.1997; Taxonomy and systematics of staphylococci indigenous to humans. In The Staphylococci in Human Disease pp. 113–137 Edited by
CrossleyK. B.,
ArcherG. L.
New York: Churchill Livingstone;
KloosW. E.,
BannermanT. L.1995; Staphylococcus and Micrococcus. Manual of Clinical Microbiology, 6th. edn, pp. 282–298 Edited by
MurrayP. R.,
BaronE. J.,
PfallerM. A.,
TenoverF. C.,
YolkenR. H.
Washington, DC: American Society for Microbiology;
KloosW. E.,
BallardD. N.,
GeorgeC. G.,
WebsterJ. A.,
HubnerR. J.,
LudwigW.,
SchleiferK.-H.,
FiedlerF.,
SchubertK.1998; Delimiting the genus Staphylococcus through description of Macrococcus caseolyticus gen. nov., comb. nov. and Macrococcus equipercicus sp. nov., Macrococcus bovicus sp. nov. and Macrococcus carouselicus sp. nov. Int J Syst Bacteriol 48:859–877
Martin de NicolasM. M.,
VindelA.,
Saez-NietoJ. A.1995; Epidemiological typing of clinically significant strains of coagulase-negative staphylococci. J Hosp Infect 29:35–43
O’DonnellA. G.,
NahaieM. R.,
GoodfellowM.,
MinnikinD. E.,
HajekV.1985; Numerical analysis of fatty acid profiles in the identification of staphylococci. J Gen Microbiol 131:2023–2033
PaiS.,
EsenN.,
PanX.,
MusserJ. M.1997; Routine rapid mycobacterium species assignment based on species-specific allelic variation in the 65-kilodalton heat shock protein gene (hsp65). Arch Pathol Lab Med 121:859–864
PilletJ.,
OrtaB.1981; Species and serotypes in coagulase-negative Staphylococci. In. Staphylococci and Staphylocoocal Infections. pp. 147–152Zentralblatt fur Bakteriologie, Mikro- biologie und Hygiene. I. Abteilung Supplemente vol. 10 Edited by
JeljaszewiczJ.,
HryniewiczW.,
SwitalskiL. M.,
SzmigielskiS.
Stuttgart New York: Gustav Fischer Verlag;
RosypalS.,
RosypalovaA.,
HorejsJ.1966; The classification of micrococci and staphylococci based on their DNA base composition and Adansonian analysis. J Gen Microbiol 44:281–292
StackebrandtE.,
GoebelB. M.1994; A place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int J Syst Bacteriol 44:846–849
SteingrubeV. A.,
GibsonJ. L.,
BrownB. A.,
ZhangY.,
WilsonR. W.,
RajagopalanM.,
WallaceR. J.Jr1995; PCR amplification and restriction endonuclease analysis of a 65-kilodalton heat shock protein gene sequence for taxonomic separation of rapidly growing mycobacteria. J Clin Microbiol 33:149–153
ThompsonJ. D.,
HigginsD. G.,
GibsonT. J.1994; clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighing, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 224673–4680
VialeA. M.,
ArakakiA. K.,
SonciniF. C.,
FerreyraR. G.1994; Evolutionary relationships among eubacterial groups as inferred from GroEL (chaperonin) sequence comparisons. Int J Syst Bacteriol 44:527–533
WayneL. G.,
BrennerD. J.,
ColwellR. R.
9 other authors 1987; International Committee on Systematic Bacteriology. Report of the ad hoc committee on reconciliation of approaches to bacterial systematics. Int J Syst Bacteriol 37:463–464
YamamotoS.,
HarayamaS.1995; PCR amplification and direct sequencing of gyrB genes with universal primers and their application to the detection and taxonomic analysis of Pseudomonas putida strains. Appl Environ Microbiol 61:1104–1109
Zakrzewska-CzerwinskaJ.,
Gaszewska-MastalarzA.,
LisB.,
GamianA.,
MordarskiM.1995; Staphylococcus pulvereri sp. nov., isolated from human and animal specimens. Int J Syst Bacteriol 45:169–172