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J Clin Microbiol. Apr 1996; 34(4): 866–869.
PMCID: PMC228907

Rapid identification of mycobacterial species by PCR amplification of hypervariable 16S rRNA gene promoter region.


A total of 0.3 to 0.4 kb of the promoter region of the 16S rRNA genes from Mycobacterium tuberculosis, M. gordonae, M. xenopi, and M. leprae was PCR amplified, cloned, and sequenced. The observed number of substitutions, insertions, and deletions exceeded those found in previously used target sequences, including the entire 16S coding region. A simple and generally applicable restriction fragment length polymorphism method that can be used to distinguish between mycobacterial species is described.

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Selected References

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  • Böddinghaus B, Rogall T, Flohr T, Blöcker H, Böttger EC. Detection and identification of mycobacteria by amplification of rRNA. J Clin Microbiol. 1990 Aug;28(8):1751–1759. [PMC free article] [PubMed]
  • Böttger EC, Hirschel B, Coyle MB. Mycobacterium genavense sp. nov. Int J Syst Bacteriol. 1993 Oct;43(4):841–843. [PubMed]
  • Compton J. Nucleic acid sequence-based amplification. Nature. 1991 Mar 7;350(6313):91–92. [PubMed]
  • Dams E, Yamada T, De Baere R, Huysmans E, Vandenberghe A, De Wachter R. Structure of 5S rRNA in actinomycetes and relatives and evolution of eubacteria. J Mol Evol. 1987;25(3):255–260. [PubMed]
  • Haas WH, Kirschner P, Ziesing S, Bremer HJ, Böttger EC. Cervical lymphadenitis in a child caused by a previously unknown mycobacterium. J Infect Dis. 1993 Jan;167(1):237–240. [PubMed]
  • Higgins DG, Sharp PM. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. [PubMed]
  • Ji YE, Colston MJ, Cox RA. Nucleotide sequence and secondary structures of precursor 16S rRNA of slow-growing mycobacteria. Microbiology. 1994 Jan;140(Pt 1):123–132. [PubMed]
  • Kapur V, Li LL, Hamrick MR, Plikaytis BB, Shinnick TM, Telenti A, Jacobs WR, Jr, Banerjee A, Cole S, Yuen KY, et al. Rapid Mycobacterium species assignment and unambiguous identification of mutations associated with antimicrobial resistance in Mycobacterium tuberculosis by automated DNA sequencing. Arch Pathol Lab Med. 1995 Feb;119(2):131–138. [PubMed]
  • Kempsell KE, Ji YE, Estrada IC, Colston MJ, Cox RA. The nucleotide sequence of the promoter, 16S rRNA and spacer region of the ribosomal RNA operon of Mycobacterium tuberculosis and comparison with Mycobacterium leprae precursor rRNA. J Gen Microbiol. 1992 Aug;138(Pt 8):1717–1727. [PubMed]
  • Kirschner P, Teske A, Schröder KH, Kroppenstedt RM, Wolters J, Böttger EC. Mycobacterium confluentis sp. nov. Int J Syst Bacteriol. 1992 Apr;42(2):257–262. [PubMed]
  • Liesack W, Pitulle C, Sela S, Stackebrandt E. Nucleotide sequence of the 16S rRNA from Mycobacterium leprae. Nucleic Acids Res. 1990 Sep 25;18(18):5558–5558. [PMC free article] [PubMed]
  • Mehra V, Sweetser D, Young RA. Efficient mapping of protein antigenic determinants. Proc Natl Acad Sci U S A. 1986 Sep;83(18):7013–7017. [PMC free article] [PubMed]
  • Meier A, Kirschner P, Schröder KH, Wolters J, Kroppenstedt RM, Böttger EC. Mycobacterium intermedium sp. nov. Int J Syst Bacteriol. 1993 Apr;43(2):204–209. [PubMed]
  • Pitulle C, Dorsch M, Kazda J, Wolters J, Stackebrandt E. Phylogeny of rapidly growing members of the genus Mycobacterium. Int J Syst Bacteriol. 1992 Jul;42(3):337–343. [PubMed]
  • Rogall T, Wolters J, Flohr T, Böttger EC. Towards a phylogeny and definition of species at the molecular level within the genus Mycobacterium. Int J Syst Bacteriol. 1990 Oct;40(4):323–330. [PubMed]
  • Ross BC, Jackson K, Yang M, Sievers A, Dwyer B. Identification of a genetically distinct subspecies of Mycobacterium kansasii. J Clin Microbiol. 1992 Nov;30(11):2930–2933. [PMC free article] [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Sela S, Clark-Curtiss JE. Cloning and characterization of the Mycobacterium leprae putative ribosomal RNA promoter in Escherichia coli. Gene. 1991 Feb 1;98(1):123–127. [PubMed]
  • Smida J, Kazda J, Stackebrandt E. Molecular-genetic evidence for the relationship of Mycobacterium leprae to slow-growing pathogenic mycobacteria. Int J Lepr Other Mycobact Dis. 1988 Sep;56(3):449–454. [PubMed]
  • Springer B, Kirschner P, Rost-Meyer G, Schröder KH, Kroppenstedt RM, Böttger EC. Mycobacterium interjectum, a new species isolated from a patient with chronic lymphadenitis. J Clin Microbiol. 1993 Dec;31(12):3083–3089. [PMC free article] [PubMed]
  • Stahl DA, Urbance JW. The division between fast- and slow-growing species corresponds to natural relationships among the mycobacteria. J Bacteriol. 1990 Jan;172(1):116–124. [PMC free article] [PubMed]
  • Telenti A, Marchesi F, Balz M, Bally F, Böttger EC, Bodmer T. Rapid identification of mycobacteria to the species level by polymerase chain reaction and restriction enzyme analysis. J Clin Microbiol. 1993 Feb;31(2):175–178. [PMC free article] [PubMed]
  • van der Vliet GM, Schukkink RA, van Gemen B, Schepers P, Klatser PR. Nucleic acid sequence-based amplification (NASBA) for the identification of mycobacteria. J Gen Microbiol. 1993 Oct;139(10):2423–2429. [PubMed]

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