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Journal of Clinical Microbiology, August 1999, p. 2667-2673, Vol. 37, No. 8
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Detection of Staphylococcus aureus and Staphylococcus epidermidis in Clinical Samples by 16S rRNA-Directed In Situ Hybridization

Vanessa Krimmer,1 Hilde Merkert,1 Christof von Eiff,2 Matthias Frosch,3 Jochen Eulert,4 Jochen F. Löhr,5 Jörg Hacker,1 and Wilma Ziebuhr1,*

Institut für Molekulare Infektionsbiologie1 and Institüt für Hygiene und Mikrobiologie,3 Universität Würzburg, and König-Ludwig-Haus, Orthopädische Klinik,4 Würzburg, and Institut für Medizinische Mikrobiologie, Universität Münster, Münster,2 Germany, and Schultheißklinik, Orthopädische Chirurgie, Zürich, Switzerland5

Received 17 November 1998/Returned for modification 1 February 1999/Accepted 29 March 1999

Staphylococcus epidermidis and Staphylococcus aureus are the most common causes of medical device-associated infections, including septicemic loosenings of orthopedic implants. Frequently, the microbiological diagnosis of these infections remains ambiguous, since at least some staphylococci have the capacity to reduce their growth rate considerably. These strains exhibit a small-colony phenotype, and often they are not detectable by conventional microbiological techniques. Moreover, clinical isolates of S. aureus and S. epidermidis adhere to polymer and metal surfaces by the generation of thick, multilayered biofilms consisting of bacteria and extracellular polysaccharides. This study reports improved detection and identification of S. aureus and S. epidermidis by an in situ hybridization method with fluorescence-labeled oligonucleotide probes specific for staphylococcal 16S rRNA. The technique has proven to be suitable for the in situ detection of staphylococci, which is illustrated by the identification of S. epidermidis in a connective tissue sample obtained from a patient with septicemic loosening of a hip arthroplasty. We also show that this technique allows the detection of intracellularly persisting bacteria, including small-colony variants of S. aureus, and the differentiation of S. epidermidis from other clinically relevant staphylococci even when they are embedded in biofilms. These results suggest that the 16S rRNA in situ hybridization technique could represent a powerful diagnostic tool for the detection and differentiation of many other fastidious microorganisms.

* Corresponding author. Mailing address: Institut für Molekulare Infektionsbiologie, Röntgenring 11, 97070 Würzburg, Germany. Phone: 49 931-312154. Fax: 49 931-312578. E-mail: w.ziebuhr{at}mail.uni-wuerzburg.de.

Journal of Clinical Microbiology, August 1999, p. 2667-2673, Vol. 37, No. 8
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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