Identification of mycobacteria by nonradioisotopic single-strand conformation polymorphism analysis.
Clinical isolates of mycobacteria were identified to species levels using nonradioisotopic single-strand conformation polymorphism (non-RI SSCP) analysis of 16S rRNA gene fragments amplified by polymerase chain reaction with primers common to all of mycobacterial species. The method is based on a hypervariable region within the 16S rRNA in mycobacteria, which is characterized by species-specific nucleotide sequences. A total of 92 mycobacterial strains (Mycobacterium tuberculosis, M. avium, M. gordonae, M. intracellulare, M. kansasii, M. chelonae, M. nonchromogenicum, M. xenopi, and unidentified strain) were studied. They were classified into nine types of pattern showing single-strand DNA bands having different mobilities. Each strain was shown in the species-specific mobility by non-RI SSCP analysis. The results of non-RI SSCP analysis were identical to those of standard biochemical methods and 16S rRNA sequencing.
Tokue
Y
Y
Clinical Laboratory Division, National Cancer Center Hospital, Tokyo, Japan.
Sugano
K
K
Noda
T
T
Saito
D
D
Shimosato
Y
Y
Ohkura
H
H
Kakizoe
T
T
Sekiya
T
T
D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D006801:Humans; D007553:Isotope Labeling; D009161:Mycobacterium / Q000145:classification / Q000235:genetics*; D018807:Polymorphism, Single-Stranded Conformational; D012329:RNA, Bacterial / Q000032:analysis*; D012336:RNA, Ribosomal, 16S / Q000032:analysis*
D017931:DNA Primers; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S