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Journal of Clinical Microbiology, January 2000, p. 246-251, Vol. 38, No. 1
Department of Pathology and Laboratory
Medicine, The University of Pennsylvania,
Philadelphia,1 and Pennsylvania State
Public Health Laboratory, Pennsylvania Department of Health,
Lionville,3 Pennsylvania, and Department
of Pathology, Baylor College of Medicine, Houston,
Texas2
Received 8 June 1999/Returned for modification 22 July
1999/Accepted 8 October 1999
We evaluated the MicroSeq 500 16S rDNA Bacterial Sequencing Kit (PE
Applied Biosystems), a 500-bp sequence-based identification system, for
its ability to identify clinical Mycobacterium isolates. The organism identity was determined by comparing the 16S rDNA sequence
to the MicroSeq database, which consists primarily of type strain
sequences. A total of 113 isolates (18 different species), previously
recovered and identified by routine methods from two clinical
laboratories, were analyzed by the MicroSeq method. Isolates with
discordant results were analyzed by hsp65 gene sequence
analysis and in some cases repeat phenotypic identification, AccuProbe rRNA hybridization (Gen-Probe, Inc., San Diego, Calif.), or
high-performance liquid chromatography of mycolic acids. For 93 (82%)
isolates, the MicroSeq identity was concordant with the previously
reported identity. For 18 (16%) isolates, the original identification
was discordant with the MicroSeq identification. Of the 18 discrepant isolates, 7 (six unique sequences) were originally misidentified by
phenotypic analysis or the AccuProbe assay but were correctly identified by the MicroSeq assay. Of the 18 discrepant isolates, 11 (seven unique sequences) were unusual species that were difficult to
identify by phenotypic methods and, in all but one case, by molecular
methods. The remaining two isolates (2%) failed definitive phenotypic
identification, but the MicroSeq assay was able to definitively
identify one of these isolates. The MicroSeq identification system is
an accurate and rapid method for the identification of
Mycobacterium spp.
0095-1137/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Sequence-Based Identification of
Mycobacterium Species Using the MicroSeq 500 16S rDNA
Bacterial Identification System
*
Corresponding author. Mailing address: Clinical
Microbiology Laboratory, Department of Pathology and Laboratory
Medicine, 4th Floor, Gates Bldg., 3400 Spruce St., Philadelphia, PA
19104-4283. Phone: (215) 662-6651. Fax: (215) 662-6655. E-mail:
jbpatel{at}mail.med.upenn.edu.
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