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Journal of Clinical Microbiology, February 2000, p. 814-817, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Rapid Identification of Bacteria in Blood Cultures by Using Fluorescently Labeled Oligonucleotide Probes

G. J. Jansen, M. Mooibroek, J. Idema, H. J. M. Harmsen, G. W. Welling,* and J. E. Degener

Department of Medical Microbiology, University of Groningen, Groningen, The Netherlands

Received 11 June 1999/Returned for modification 10 September 1999/Accepted 8 November 1999

The applicability of whole-cell hybridization for the identification of pathogenic bacteria in blood from septic patients was examined. Oligonucleotide probes, fluorescently labeled with fluorescein isothiocyanate, directed against the variable regions of the 16S rRNAs of the following bacterial species and/or genera were used: Streptococcus spp., Enterococcus faecalis, Staphylococcus aureus, coagulase-negative staphylococci (CoNS), Escherichia coli, Pseudomonas aeruginosa, and the Enterobacteriaceae family. A probe specific for the rRNAs of almost all bacteria and its complementary, reversed counterpart was used as positive and negative control, respectively. The probes were used in conjunction with a fast and simple-to-use protocol for whole-cell hybridization. This protocol yields an identification after 25 to 45 min, depending on whether the bacterium is gram positive or gram negative. A total of 182 blood samples which tested positive in a blood culture machine were investigated. All probes except for the ones for S. aureus and the CoNS showed sensitivities and specificities of 1.000. It was concluded that whole-cell hybridization is well suited for the fast screening of septic blood containing streptococci and/or enterococci or gram-negative rods.

* Corresponding author. Mailing address: Department of Medical Microbiology, University of Groningen, Hanzeplein 1, Entrance 21, 2nd floor, 9713 GZ, Groningen, The Netherlands. Phone: 31-503633513. Fax: 31-503633528. E-mail: g.w.welling{at}med.rug.nl.

Journal of Clinical Microbiology, February 2000, p. 814-817, Vol. 38, No. 2
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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