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Journal of Clinical Microbiology, July 2008, p. 2241-2246, Vol. 46, No. 7
0095-1137/08/$08.00+0     doi:10.1128/JCM.00347-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Multiplex Real-Time PCR Assay for Rapid Identification of Mycobacterium tuberculosis Complex Members to the Species Level{triangledown}

Benjamin A. Pinsky1* and Niaz Banaei1,2

Department of Pathology, Stanford University School of Medicine, Stanford, California 94305,1 Clinical Microbiology Laboratory, Stanford Hospital and Clinics, Palo Alto, California 943042

Received 19 February 2008/ Returned for modification 3 May 2008/ Accepted 20 May 2008

The species identification of members of the Mycobacterium tuberculosis complex is critical to the timely initiation of both appropriate antibiotic therapy and proper public health control measures. However, the current commercially available molecular assays identify mycobacteria only to the complex level and are unable to differentiate M. tuberculosis from the closely related M. bovis and M. bovis BCG. We describe here a rapid and robust two-step, multiplex, real-time PCR assay based on genomic deletions to definitively identify M. tuberculosis, M. bovis, M. bovis BCG, and other members of the complex. When tested against a panel of well-characterized mycobacterial reference strains, the assay was both sensitive and specific, correctly identifying all strains. We applied this assay to 60 clinical isolates previously identified as M. tuberculosis complex and found 57 M. tuberculosis isolates and 3 M. bovis BCG isolates from patients who had received intravesical BCG. Furthermore, analysis of 15 clinical specimens previously identified as M. bovis by spoligotyping revealed an isolate of M. tuberculosis that had been misidentified. We propose that this assay will allow the routine identification of M. tuberculosis complex members in the clinical laboratory.

* Corresponding author. Mailing address: Stanford University School of Medicine, Department of Pathology, 300 Pasteur Drive, Lane 235, Stanford, CA 94305-5324. Phone: (650) 723-5252. Fax: (650) 725-6902. E-mail: bpinsky{at}stanford.edu

{triangledown} Published ahead of print on 28 May 2008.

Journal of Clinical Microbiology, July 2008, p. 2241-2246, Vol. 46, No. 7
0095-1137/08/$08.00+0     doi:10.1128/JCM.00347-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Richardson, E. T., Samson, D., Banaei, N. (2009). Rapid Identification of Mycobacterium tuberculosis and Nontuberculous Mycobacteria by Multiplex, Real-Time PCR. J. Clin. Microbiol. 47: 1497-1502 [Abstract] [Full Text]  


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