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Journal of Clinical Microbiology, January 1999, p. 49-55, Vol. 37, No. 1
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Mycobacterium Species Identification and Rifampin Resistance Testing with High-Density DNA Probe Arrays

A. Troesch,1,* H. Nguyen,2 C. G. Miyada,2 S. Desvarenne,1 T. R. Gingeras,2 P. M. Kaplan,2 P. Cros,1 and C. Mabilat1

bioMérieux, 69280 Marcy-L'Etoile, France,1 and Affymetrix, Santa Clara, California2

Received 27 July 1998/Returned for modification 3 September 1998/Accepted 15 October 1998

Species identification within the genus Mycobacterium and subsequent antibiotic susceptibility testing still rely on time-consuming, culture-based methods. Despite the recent development of DNA probes, which greatly reduce assay time, there is a need for a single platform assay capable of answering the multitude of diagnostic questions associated with this genus. We describe the use of a DNA probe array based on two sequence databases: one for the species identification of mycobacteria (82 unique 16S rRNA sequences corresponding to 54 phenotypical species) and the other for detecting Mycobacterium tuberculosis rifampin resistance (rpoB alleles). Species identification or rifampin resistance was determined by hybridizing fluorescently labeled, amplified genetic material generated from bacterial colonies to the array. Seventy mycobacterial isolates from 27 different species and 15 rifampin-resistant M. tuberculosis strains were tested. A total of 26 of 27 species were correctly identified as well as all of the rpoB mutants. This parallel testing format opens new perspectives in terms of patient management for bacterial diseases by allowing a number of genetic tests to be simultaneously run.

* Corresponding author. Mailing address: Affymetrix, 3380 Central Expressway, Santa Clara, CA 95051. Phone: (408) 731 5572. Fax: (408) 481 0435. E-mail: alain_troesch{at}affymetrix.com.

Journal of Clinical Microbiology, January 1999, p. 49-55, Vol. 37, No. 1
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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