This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Richardson, E. T.
Right arrow Articles by Banaei, N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Richardson, E. T.
Right arrow Articles by Banaei, N.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, May 2009, p. 1497-1502, Vol. 47, No. 5
0095-1137/09/$08.00+0     doi:10.1128/JCM.01868-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Rapid Identification of Mycobacterium tuberculosis and Nontuberculous Mycobacteria by Multiplex, Real-Time PCR{triangledown}

E. T. Richardson,1 D. Samson,2 and N. 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 26 September 2008/ Returned for modification 15 November 2008/ Accepted 5 March 2009

The rapid identification of mycobacteria from culture is of primary importance for the administration of empirical antibiotic therapy and for the implementation of public health measures, yet there are few commercially available assays that can easily and accurately identify the mycobacteria in culture in a timely manner. Here we report on the development of a multiplex, real-time PCR assay that can identify 93% of the pathogenic mycobacteria in our laboratory in two parallel reactions. The mycobacteria identified by this assay include the Mycobacterium tuberculosis complex (MTC), the M. avium complex (MAC), the M. chelonae-M. abscessus group (MCAG), the M. fortuitum group (MFG), and M. mucogenicum. The primer targets included the 16S rRNA gene and the internal transcribed spacer. The assay was initially validated with a repository of reference strains and was subsequently tested with 314 clinical cultures identified by the AccuProbe assay or high-performance liquid chromatography. Of the 314 cultures tested, multiplex, real-time PCR produced congruent results for 99.8% of the 1,559 targets evaluated. The sensitivity and the specificity were each 99% or greater for MTC (n = 96), MAC (n = 97), MCAG (n = 68), and M. mucogenicum (n = 9) and 95% and 100%, respectively, for MFG (n = 19). We conclude that this multiplex, real-time PCR assay is a useful diagnostic tool for the rapid and accurate identification of MTC and clinically relevant nontuberculous mycobacteria.

* Corresponding author. Mailing address: Stanford University School of Medicine, Department of Pathology, 3375 Hillview Avenue, Room 1602, Palo Alto, CA 94304. Phone: (650) 736-8052. Fax: (650) 725-5671. E-mail: niazbanaei{at}stanford.edu

{triangledown} Published ahead of print on 18 March 2009.

Journal of Clinical Microbiology, May 2009, p. 1497-1502, Vol. 47, No. 5
0095-1137/09/$08.00+0     doi:10.1128/JCM.01868-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»