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Development of a real‐time PCR‐based method for rapid differential identification of Mycobacterium species
Author(s) -
Lim S.Y.,
Kim B.J.,
Lee M.K.,
Kim K.
Publication year - 2008
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2007.02278.x
Subject(s) - rpob , biology , mycobacterium , nontuberculous mycobacteria , polymerase chain reaction , species complex , identification (biology) , primer (cosmetics) , mycobacterium fortuitum , genus , microbiology and biotechnology , real time polymerase chain reaction , computational biology , gene , genetics , 16s ribosomal rna , bacteria , zoology , ecology , phylogenetic tree , chemistry , organic chemistry
Aims: To develop a real‐time PCR method for rapid differential identification of many clinically important mycobacteria to the species level. Methods and 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. Significance and Impact of the Study: This real‐time PCR assay provides a useful tool for the rapid differentiation of most clinically important Mycobacterium species.