Affiliation:
1. Department of Pathology, Stanford University School of Medicine, Stanford, California 94305
2. Clinical Microbiology Laboratory, Stanford Hospital and Clinics, Palo Alto, California 94304
Abstract
ABSTRACT
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.
Publisher
American Society for Microbiology
Cited by
76 articles.
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