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Applied and Environmental Microbiology, January 2000, p. 375-382, Vol. 66, No. 1
0099-2240/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Quantification of Different Eubacterium spp. in Human Fecal Samples with Species-Specific 16S rRNA-Targeted Oligonucleotide Probes

Andreas Schwiertz, Gwenaelle Le Blay, and Michael Blaut*

Deutsches Institut für Ernährungsforschung, Abteilung Gastrointestinale Mikrobiologie, 14558 Bergholz-Rehbrücke, Germany

Received 10 August 1999/Accepted 15 October 1999

Species-specific 16S rRNA-targeted, Cy3 (indocarbocyanine)-labeled oligonucleotide probes were designed and validated to quantify different Eubacterium species in human fecal samples. Probes were directed at Eubacterium barkeri, E. biforme, E. contortum, E. cylindroides (two probes), E. dolichum, E. hadrum, E. lentum, E. limosum, E. moniliforme, and E. ventriosum. The specificity of the probes was tested with the type strains and a range of common intestinal bacteria. With one exception, none of the probes showed cross-hybridization under stringent conditions. The species-specific probes were applied to fecal samples obtained from 12 healthy volunteers. E. biforme, E. cylindroides, E. hadrum, E. lentum, and E. ventriosum could be determined. All other Eubacterium species for which probes had been designed were under the detection limit of 107 cells g (dry weight) of feces-1. The cell counts obtained are essentially in accordance with the literature data, which are based on colony counts. This shows that whole-cell in situ hybridization with species-specific probes is a valuable tool for the enumeration of Eubacterium species in feces.


* Corresponding author. Mailing address: Deutsches Institut für Ernährungsforschung, Abteilung Gastrointestinale Mikrobiologie, Arthur-Scheunert-Allee 114-116, D-14558 Bergholz-Rehbrücke, Germany. Phone: 49-33200-88470. Fax: 49-33200-88407. E-mail: blaut{at}www.dife.de.


Applied and Environmental Microbiology, January 2000, p. 375-382, Vol. 66, No. 1
0099-2240/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



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