Development of a real-time PCR-based method for rapid differential identification of Mycobacterium species.
OBJECTIVE
To develop a real-time PCR method for rapid differential identification of many clinically important mycobacteria to the species level.


RESULTS
Eighteen Mycobacterium species that are considered clinically important were targeted for the identification. One primer pair and 21 pairs of hybridization probes (HybProbes) specific for the genus, species or complex were designed based on the rpoB gene sequences of mycobacteria. Twenty-five different Mycobacterium reference species were tested. In a single round of real-time PCR, all the nontuberculous mycobacteria (NTM) species tested were identified at the genus level and 16 of the 18 targeted species were differentially identified to the species or complex level during the amplification cycles; subsequent melting curve analysis allowed the specific identification of all the target species at the species or complex level without cross-reactivity with the other species.


CONCLUSIONS
The developed real-time PCR assay rapidly identifies the NTM at the genus level and 18 clinically important Mycobacterium species at the species or complex level.


CONCLUSIONS
This real-time PCR assay provides a useful tool for the rapid differentiation of most clinically important Mycobacterium species.
Lim
S Y
SY
Division of Pulmonary and Critical Care Medicine, Department of Medicine, Kangbuk Samsung Hospital, Sungkyunkwan University School of Medicine, Seoul, South Korea.
Kim
B-J
BJ
Lee
M-K
MK
Kim
K
K
D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D017931:DNA Primers / Q000235:genetics; D012321:DNA-Directed RNA Polymerases; D005784:Gene Amplification; D006801:Humans; D025202:Molecular Diagnostic Techniques; D015335:Molecular Probes / Q000235:genetics; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D009164:Mycobacterium Infections / Q000175:diagnosis / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D044366:Transition Temperature
D001426:Bacterial Proteins; D017931:DNA Primers; D015335:Molecular Probes; C505805:rpoB protein, Mycobacterium tuberculosis; D012321:DNA-Directed RNA Polymerases