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J Med Microbiol 54 (2005), 1127-1131; DOI: 10.1099/jmm.0.46129-0
© 2005 Society for General Microbiology
ISSN 0022-2615

Microarray-based pncA genotyping of pyrazinamide-resistant strains of Mycobacterium tuberculosis

Steven Denkin1, Dmitriy Volokhov2, Vladimir Chizhikov2 and Ying Zhang1

1Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland 21205, USA 2Center for Biologics Evaluation and Research, Food and Drug Administration, Kensington, MD 20895, USA

Correspondence Ying Zhang yzhang{at}jhsph.edu

Received 19 April 2005
Accepted 12 August 2005

Drug-resistant Mycobacterium tuberculosis poses a significant threat to the treatment of tuberculosis (TB). The current susceptibility testing for the first-line TB drug pyrazinamide (PZA) is not only time-consuming but also difficult, due to the requirement for acid pH for drug activity. Predominantly, resistance to PZA in M. tuberculosis is caused by mutations in the pncA gene, and the detection of pncA mutations can be an indicator of PZA resistance. In this study, the use of a previously developed microarray method for the rapid detection of PZA-resistant M. tuberculosis based on identifying mutations in the pncA gene was evaluated. Microarray analysis was performed in a blind manner on 33 clinical isolates of M. tuberculosis for which the sequence of the pncA gene had not previously been determined. The results showed that all mutations in PZA-resistant strains identified by DNA sequencing could be unambiguously detected by the microarray method. It is concluded that the microarray method is a valuable tool for the rapid screening and genetic identification of potential PZA-resistant M. tuberculosis strains.

Abbreviations: POA, pyrazinoic acid; PZA, pyrazinamide; PZase, pyrazinamidase; ssRNA, single-stranded RNA; TB, tuberculosis.




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