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Journal of Clinical Microbiology, May 2001, p. 1763-1770, Vol. 39, No. 5
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.5.1763-1770.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Classification and Identification of Enterococci: a Comparative Phenotypic, Genotypic, and Vibrational Spectroscopic Study

C. Kirschner,1,* K. Maquelin,2 P. Pina,3 N. A. Ngo Thi,1 L.-P. Choo-Smith,2 G. D. Sockalingum,4 C. Sandt,4 D. Ami,5 F. Orsini,5 S. M. Doglia,5 P. Allouch,3 M. Mainfait,4 G. J. Puppels,2 and D. Naumann1,*

Robert Koch Institute, Biophysical Structure Analyses, 13353 Berlin, Germany1; Laboratory for Intensive Care Research and Optical Spectroscopy, Erasmus University Rotterdam, and Department of General Surgery 10M, University Hospital Rotterdam Dijkzigt, 3015 GD Rotterdam, The Netherlands2; Service d'Hygiène Hospitalière, Centre Hospitalier de Versailles, 78157 Le Chesnay Cedex,3 and Unité MéDIAN, CNRS FRE 2141, IFR 53, UFR de Pharmacie, Université de Reims Champagne-Ardenne, 51096 Reims Cedex,4 France; and UdR INFM Milano-Bicocca, Dipartimento di Biotecnologie e Bioscienze, 20126 Milan, Italy5

Received 6 November 2000/Returned for modification 29 January 2001/Accepted 23 February 2001

Rapid and accurate identification of enterococci at the species level is an essential task in clinical microbiology since these organisms have emerged as one of the leading causes of nosocomial infections worldwide. Vibrational spectroscopic techniques (infrared [IR] and Raman) could provide potential alternatives to conventional typing methods, because they are fast, easy to perform, and economical. We present a comparative study using phenotypic, genotypic, and vibrational spectroscopic techniques for typing a collection of 18 Enterococcus strains comprising six different species. Classification of the bacteria by Fourier transform (FT)-IR spectroscopy in combination with hierarchical cluster analysis revealed discrepancies for certain strains when compared with results obtained from automated phenotypic systems, such as API and MicroScan. Further diagnostic evaluation using genotypic methods---i.e., PCR of the species-specific ligase and glycopeptide resistance genes, which is limited to the identification of only four Enterococcus species and 16S RNA sequencing, the "gold standard" for identification of enterococci---confirmed the results obtained by the FT-IR classification. These results were later reproduced by three different laboratories, using confocal Raman microspectroscopy, FT-IR attenuated total reflectance spectroscopy, and FT-IR microspectroscopy, demonstrating the discriminative capacity and the reproducibility of the technique. It is concluded that vibrational spectroscopic techniques have great potential as routine methods in clinical microbiology.

* Corresponding author. Mailing address for D. Naumann: P34, Robert Koch Institute, Biophysical Structure Analyses, Nordufer 20, 13353 Berlin, Germany. Phone: 49-30-45472259. Fax: 49-30-45472606. E-mail: NaumannD{at}rki.de. Mailing address for C. Kirschner: P34, Robert Koch Institute, Biophysical Structure Analyses, Nordufer 20, 13353 Berlin, Germany. Phone: 49-30-45472519. Fax: 49-30-45472606. E-mail: KirschnerC{at}rki.de.

Journal of Clinical Microbiology, May 2001, p. 1763-1770, Vol. 39, No. 5
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.5.1763-1770.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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