Differential identification of Mycobacterium tuberculosis complex and nontuberculous mycobacteria by duplex PCR assay using the RNA polymerase gene (rpoB).
A novel duplex PCR method that can amplify the 235- and 136-bp rpoB DNAs of Mycobacterium tuberculosis complex and nontuberculous mycobacteria (NTM), respectively, with two different sets of primers was used to differentially identify 44 reference strains and 379 clinical isolates of mycobacteria in a single-step assay. Showing 100% sensitivity and specificity, the duplex PCR method could clearly differentiate M. tuberculosis complex and NTM strains. In addition, restriction fragment length polymorphism analysis and direct sequencing of the amplicon of NTM could be used to supplement species identification.
Kim
Bum-Joon
BJ
Department of Microbiology and Cancer Research Institute, Institute of Endemic Diseases, SNUMRC, Seoul National University College of Medicine, Seoul 137-140, Korea.
Hong
Seong-Karp
SK
Lee
Keun-Hwa
KH
Yun
Yeo-Jun
YJ
Kim
Eui-Chong
EC
Park
Young-Gil
YG
Bai
Gil-Han
GH
Kook
Yoon-Hoh
YH
D001483:Base Sequence; D002874:Chromosome Mapping; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D006801:Humans; D009161:Mycobacterium / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D009164:Mycobacterium Infections / Q000175:diagnosis*; D009169:Mycobacterium tuberculosis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit