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Journal of Clinical Microbiology, April 2000, p. 1676-1678, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Identification of Coryneform Bacterial Isolates by Ribosomal DNA Sequence Analysis

Yi-Wei Tang,1,* Alexander Von Graevenitz,2 Michael G. Waddington,3 Marlene K. Hopkins,4 Douglas H. Smith,3,5 Haijing Li,1 Christopher P. Kolbert,4 Stacy O. Montgomery,5 and David H. Persing6

Departments of Medicine and Pathology, Vanderbilt University School of Medicine, Nashville, Tennessee 372321; Department of Medical Microbiology, University of Zurich, Zurich, CH-8028 Switzerland2; MIDI Labs, Inc., Newark, Delaware 197133; Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota 559054; Perkin-Elmer Biosystems, Foster City, California 944045; and Corixa Corporation and the Infectious Disease Research Institute, Seattle, Washington 981046

Received 22 November 1999/Returned for modification 4 January 2000/Accepted 26 January 2000

Identification of coryneform bacteria to the species level is important in certain circumstances for differentiating contamination and/or colonization from infection, which influences decisions regarding clinical intervention. However, methods currently used in clinical microbiology laboratories for the species identification of coryneform bacteria are often inadequate. We evaluated the MicroSeq 500 16S bacterial sequencing kit (Perkin-Elmer Biosystems, Foster City, Calif.), which is designed to sequence the first 527 bp of the 16S rRNA gene for bacterial identification, by using 52 coryneform gram-positive bacilli from clinical specimens isolated from January through June 1993 at the Mayo Clinic. Compared to conventional and supplemented phenotypic methods, MicroSeq provided concordant results for identification to the genus level for all isolates. At the species level, MicroSeq provided concordant results for 27 of 42 (64.3%) Corynebacterium isolates and 5 of 6 (83.3%) Corynebacterium-related isolates, respectively. Within the Corynebacterium genus, MicroSeq gave identical species-level identifications for the clinically significant Corynebacterium diphtheriae (4 of 4) and Corynebacterium jeikeium (8 of 8), but it identified only 50.0% (15 of 30) of other species (P < 0.01). Four isolates from the genera Arthrobacter, Brevibacterium, and Microbacterium, which could not be identified to the species level by conventional methods, were assigned a species-level identification by MicroSeq. The total elapsed time for running a MicroSeq identification was 15.5 to 18.5 h. These data demonstrate that the MicroSeq 500 16S bacterial sequencing kit provides a potentially powerful method for the definitive identification of clinical coryneform bacterium isolates.

* Corresponding author. Mailing address: A3310 MCN, Division of Infectious Diseases, Vanderbilt University Medical Center, Nashville, TN 37232-2605. Phone: (615) 322-2035. Fax: (615) 343-6160. E-mail: yiwei.tang{at}vanderbilt.edu.

Journal of Clinical Microbiology, April 2000, p. 1676-1678, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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