Open AccessRapid Identification of Mycobacterium tuberculosis and Nontuberculous Mycobacteria by Multiplex, Real-Time PCR
Author(s) -
Eugene T Richardson,
Divinia Samson,
Niaz Banaei
Publication year - 2009
Publication title -
journal of clinical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.349
H-Index - 255
eISSN - 1070-633X
pISSN - 0095-1137
DOI - 10.1128/jcm.01868-08
Subject(s) - nontuberculous mycobacteria , multiplex , microbiology and biotechnology , mycobacterium , multiplex polymerase chain reaction , mycobacterium chelonae , mycobacterium tuberculosis complex , mycobacterium fortuitum , mycobacterium tuberculosis , mycobacterium abscessus , tuberculosis , biology , polymerase chain reaction , medicine , gene , pathology , bioinformatics , genetics
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 theMycobacterium tuberculosis complex (MTC), theM. avium complex (MAC), theM. chelonae -M .abscessus group (MCAG), theM. fortuitum group (MFG), andM. 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), andM. 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.