Differentiation of Mycobacterial Species by PCR-Restriction Analysis of DNA (342 Base Pairs) of the RNA Polymerase Gene ( rpoB )
Open Access
- 1 June 2001
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 39 (6), 2102-2109
- https://doi.org/10.1128/jcm.39.6.2102-2109.2001
Abstract
PCR amplification-restriction analysis (PRA) of rpoB DNA (342 bp), which comprises the Rif r region, was used for the differential identification of 49 mycobacteria. The DNA had been used previously for the identification of mycobacterial species by comparative sequence analysis (B. J. Kim et al., J. Clin. Microbiol. 37:1714–1720, 1999). Digestion with four restriction enzymes ( Hae III, Hin dII, Mva I, and Acc II), which were selected on the basis of rpoB DNA sequences, generated distinctive PRA patterns that allowed not only the reference strains but also the clinical isolates of mycobacteria to be distinguished. Both rapidly and slowly growing mycobacteria were distinctly differentiated by Hae III digestion of the amplified rpoB DNA. By Hin dII digestion the Mycobacterium tuberculosis complex was distinguished from the other mycobacteria. Furthermore, six subspecies of Mycobacterium kansasii (subspecies I to VI) as well as the closely related Mycobacterium gastri , and other closely related species, were distinguished by simultaneous digestion of Mva I and Acc II. According to the rpoB PRA scheme, 240 strains of clinical isolates could be identified. It was also possible to detect and identify M. tuberculosis directly from sputa and bronchoalveolar lavage specimens. These results suggest that PRA of rpoB DNA is a simple and feasible method not only for the differentiation of culture isolates but also for the rapid detection and identification of pathogenic mycobacteria in primary clinical specimens.Keywords
This publication has 17 references indexed in Scilit:
- Incidence and Clinical Implications of Isolation of Mycobacterium kansasii: Results of a 5-Year, Population-Based StudyAnnals of Internal Medicine, 1998
- Demonstration of Mycobacterium kansasii species heterogeneity by the amplification of the 16S-23S spacer regionJournal of Medical Microbiology, 1995
- Restriction fragment length polymorphisms of 16S rRNA genes in the differentiation of fast-growing mycobacterial speciesFEMS Microbiology Letters, 1994
- Nucleotide Sequence Comparison of the Mycobacterial dnaJ Gene and PCR-Restriction Fragment Length Polymorphism Analysis for Identification of Mycobacterial SpeciesInternational Journal of Systematic and Evolutionary Microbiology, 1994
- Rapid, amplification-based fingerprinting of Mycobacterium tuberculosisJournal of General Microbiology, 1993
- Rapid characterization of Mycobacterium fortuitum-chelonei complex by restriction fragment length polymorphism of ribosomal RNA genesFEMS Microbiology Letters, 1991
- Rapid characterization ofMycobacterium fortuitum-cheloneicomplex by restriction fragment length polymorphism of ribosomal RNA genesFEMS Microbiology Letters, 1991
- Towards a Phylogeny and Definition of Species at the Molecular Level within the Genus MycobacteriumInternational Journal of Systematic and Evolutionary Microbiology, 1990
- IS6110, an IS-like element ofMycobacterium tuberculosiscomplexNucleic Acids Research, 1990
- The occurrence of Mycobacterium kansasii in tapwaterTubercle, 1980