1887

Abstract

Polymerase chain reaction (PCR) identification assays were designed for eight major species of coagulase-negative staphylococci (CNS) on the basis of three variable regions found in the 16S rRNA gene. The PCR assays were tested with 41 staphylococcal strains representing the diversity of staphylococci defined by classical biotyping schemes. Each PCR result was compared with species-specific polymorphism in and around the 16S rRNA gene (i.e., 16S ribotype) and the phenotypic identification of the strain in a miniaturised biochemical test gallery (bioMérieux ATB 32 Staph). Twenty-six of the 41 strains were identified by PCR as belonging to one of the eight species for which primers had been designed and none of the remaining strains was misidentified. For 22 of the 26 strains there was complete agreement between the PCR identification, 16S ribotype and ATB identification. For the remaining four strains there was agreement between PCR identification and 16S ribotype. Two National Collection of Type Culture strains were re-assigned to different species and 10 previously unassigned strains were formally speciated for the first time. These PCR assays are suitable for rapid and definitive speciation of CNS.

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1997-01-01
2022-08-13
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References

  1. Kloos W. E., Bannerman T. L. Update on clinical significance of coagulase-negative staphylococci. Clin Microbiol Rev 1994; 7:117–140
    [Google Scholar]
  2. Baird-Parker A. C. A classification of Micrococci and Staphylococci based on physiological and biochemical tests. J Gen Microbiol 1963; 30:409–427
    [Google Scholar]
  3. Kloos W. E., Schleifer K. H. Simplified scheme for routine identification of human Staphylococcus species. J Clin Microbiol 1975; 1:82–88
    [Google Scholar]
  4. Marples R. R. Taxonomic studies of Staphylococci and Micrococci. Zentralble Bakteriol Suppl 1981; 10:9–13
    [Google Scholar]
  5. Marples R. R., Richardson J. F. Evaluation of a micromethod gallery (API Staph) for the identification of staphylococci and micrococci. J Clin Pathol 1982; 35:650–656
    [Google Scholar]
  6. Freney J., Brun Y., Bes M. Staphylococcus lugdunensis sp. nov. and Staphylococcus schleiferi sp. nov., two species from human clinical specimens. Int J Syst Bacteriol 1988; 38:168–172
    [Google Scholar]
  7. Zimmerman R. J., Kloos W. E. Comparative zone electrophoresis of esterases of Staphylococcus species isolated from mammalian skin. Can J Microbiol 1976; 22:771–779
    [Google Scholar]
  8. Clink J., Pennington T. H. Staphylococcal whole-cell polypeptide analysis: evaluation as a taxonomic and typing tool. J Med Microbiol 1987; 23:41–44
    [Google Scholar]
  9. Kotilainen P., Huovinen P., Eerola E. Application of gas-liquid chromatographic analysis of cellular fatty acids for species identification and typing of coagulase-negative Staphylococci. J Clin Microbiol 1991; 29:315–322
    [Google Scholar]
  10. Schleifer K. H., Kroppenstedt R. M. Chemical and molecular classifcation of staphylococci. J Appl Bacteriol Symp Suppl 1990; 19:9S–24S
    [Google Scholar]
  11. Grimont F., Grimont P. A. D. Ribosomal ribonucleic acid gene restriction patterns as potential taxonomic tools. Ann Inst Pasteur Microbiol 1986; 137B:165–175
    [Google Scholar]
  12. De Buyser M.-L., Morvan A., Aubert S., Dilasser F., El Solh N. Evaluation of a ribosomal RNA gene probe for the identification of species and subspecies within the genus Staphylococcus. J Gen Microbiol 1992; 138:889–899
    [Google Scholar]
  13. Thomson-Carter F. M., Carter P. E., Pennington T. H. Differentiation of staphylococcal species and strains by ribosomal RNA gene restriction patterns. J Gen Microbiol 1989; 135:2093–2097
    [Google Scholar]
  14. Bialkowska-Hobrzanska H., Harry V., Jaskot D., Hammemberg O. Typing of coagulase-negative staphylococci by Southern hybridization of chromosomal DNA fingerprints using a ribosomal RNA probe. Pur J Clin Microbiol Infect Dis 1990; 9:588–594
    [Google Scholar]
  15. Wada A., Ohta H., Kulthanan K., Hiramatsu K. Molecular cloning and mapping of 16S-23S rRNA gene complexes of Staphylococcus aureus. J Bacteriol 1993; 175:7483–7487
    [Google Scholar]
  16. Zakrzewska-Czerwiska J., Gaszewska-Mastalarz A., Pulverer G., Mordarski M. Identification of Staphylococcus epidermidis using a 16S rRNA-directed oligonucleotide probe. FEMS Microbiol Lett 1992; 100:51–58
    [Google Scholar]
  17. Hedin G. Comparison of genotypic and phenotypic methods for species identification of coagulase-negative staphylococcal isolates from blood cultures. APMIS 1994; 102:855–864
    [Google Scholar]
  18. Brun Y., Bes M., Boeufgras J. M. International collaborative evaluation of the ATB 32 Staph gallery for identification of the Staphylococcus species. Int J Med Microbiol 1990; 273:319–326
    [Google Scholar]
  19. Wilson K. Preparation of genomic DNA from bacteria. In Current protocols in molecular biology New York: John Wiley and Sons Inc; 1987 unit 2.4.1
    [Google Scholar]
  20. Baquar N., Bumens A., Stanley J. Comparative evaluation of molecular typing of strains from a national epidemic due to Salmonella brandenburg by rRNA gene and IS200 probes and Pulsed-Field Gel Electrophoresis. J Clin Microbiol 1994; 32:1876–1880
    [Google Scholar]
  21. Sambrook J., Fritsch E. F., Maniatis T. Molecular cloning. A laboratory manual. 2nd edn Cold Spring Harbor, NY: Cold Spring Harbor Laboratory; 1989
    [Google Scholar]
  22. Lucas K., Busch M., Mossinger S., Thompson J. A. An improved microcomputer programme for finding gene- or family-specific oligonucleotides suitable as primers for polymerase chain reactions or as probes. Comput Appl Biosci 1991; 7:525–529
    [Google Scholar]
  23. Higgins D. G., Sharp P. M. Fast and sensitive multiple sequence alignments on a microcomputer. Comput Appl Biosci 1989; 5:151–153
    [Google Scholar]
  24. Cookson B. D., Stapleton P., Ludlam H. Ribotyping of coagulase-negative staphylococci. J Med Microbiol 1992; 36:414–419
    [Google Scholar]
  25. Schumacher-Perdreau F. Clinical significance and laboratory diagnosis of coagulase-negative Staphylococci. Clin Microbiol Newslett 1991; 13:13
    [Google Scholar]
  26. Marrie T. J., Kwan C., Noble A., West A., Duffield L. Staphylococcus saprophyticus as a cause of urinary tract infections. J Clin Micrbiol 1982; 16:427–431
    [Google Scholar]
  27. Fleurette J., Brun Y., Coulet M., Forey F. Infections caused by coagulase-negative staphylococci other than S. epidermidis and S. saprophyticus. Zentralbl Bakteriol Suppl 1987; 16:195–208
    [Google Scholar]
  28. Kleeman K. T., Bannerman T. L., Kloos W. E. Species distribution of coagulase-negative staphylococcal isolates at a community hospital and implications for selection of staphylococcal identification procedures. J Clin Microbiol 1993; 31:1318–1321
    [Google Scholar]
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