• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
J Clin Microbiol. Feb 1993; 31(2): 175–178.
PMCID: PMC262730

Rapid identification of mycobacteria to the species level by polymerase chain reaction and restriction enzyme analysis.

Abstract

A method for the rapid identification of mycobacteria to the species level was developed on the basis of evaluation by the polymerase chain reaction (PCR) of the gene encoding for the 65-kDa protein. The method involves restriction enzyme analysis of PCR products obtained with primers common to all mycobacteria. Using two restriction enzymes, BstEII and HaeIII, medically relevant and other frequent laboratory isolates were differentiated to the species or subspecies level by PCR-restriction enzyme pattern analysis. PCR-restriction enzyme pattern analysis was performed on isolates (n = 330) from solid and fluid culture media, including BACTEC, or from frozen and lyophilized stocks. The procedure does not involve hybridization steps or the use of radioactivity and can be completed within 1 working day.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (949K), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • 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, Teske A, Kirschner P, Bost S, Chang HR, Beer V, Hirschel B. Disseminated "Mycobacterium genavense" infection in patients with AIDS. Lancet. 1992 Jul 11;340(8811):76–80. [PubMed]
  • Buck GE, O'Hara LC, Summersgill JT. Rapid, simple method for treating clinical specimens containing Mycobacterium tuberculosis to remove DNA for polymerase chain reaction. J Clin Microbiol. 1992 May;30(5):1331–1334. [PMC free article] [PubMed]
  • Butler WR, Jost KC, Jr, Kilburn JO. Identification of mycobacteria by high-performance liquid chromatography. J Clin Microbiol. 1991 Nov;29(11):2468–2472. [PMC free article] [PubMed]
  • Fiss EH, Chehab FF, Brooks GF. DNA amplification and reverse dot blot hybridization for detection and identification of mycobacteria to the species level in the clinical laboratory. J Clin Microbiol. 1992 May;30(5):1220–1224. [PMC free article] [PubMed]
  • Floyd MM, Silcox VA, Jones WD, Jr, Butler WR, Kilburn JO. Separation of Mycobacterium bovis BCG from Mycobacterium tuberculosis and Mycobacterium bovis by using high-performance liquid chromatography of mycolic acids. J Clin Microbiol. 1992 May;30(5):1327–1330. [PMC free article] [PubMed]
  • Fries JW, Patel RJ, Piessens WF, Wirth DF. Genus- and species-specific DNA probes to identify mycobacteria using the polymerase chain reaction. Mol Cell Probes. 1990 Apr;4(2):87–105. [PubMed]
  • Gubler JG, Salfinger M, von Graevenitz A. Pseudoepidemic of nontuberculous mycobacteria due to a contaminated bronchoscope cleaning machine. Report of an outbreak and review of the literature. Chest. 1992 May;101(5):1245–1249. [PubMed]
  • Hance AJ, Grandchamp B, Lévy-Frébault V, Lecossier D, Rauzier J, Bocart D, Gicquel B. Detection and identification of mycobacteria by amplification of mycobacterial DNA. Mol Microbiol. 1989 Jul;3(7):843–849. [PubMed]
  • Kirschner P, Böttger EC. Microheterogeneity within rRNA of Mycobacterium gordonae. J Clin Microbiol. 1992 Apr;30(4):1049–1050. [PMC free article] [PubMed]
  • Kusunoki S, Ezaki T, Tamesada M, Hatanaka Y, Asano K, Hashimoto Y, Yabuuchi E. Application of colorimetric microdilution plate hybridization for rapid genetic identification of 22 Mycobacterium species. J Clin Microbiol. 1991 Aug;29(8):1596–1603. [PMC free article] [PubMed]
  • Kwok S, Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. [PubMed]
  • Musial CE, Tice LS, Stockman L, Roberts GD. Identification of mycobacteria from culture by using the Gen-Probe Rapid Diagnostic System for Mycobacterium avium complex and Mycobacterium tuberculosis complex. J Clin Microbiol. 1988 Oct;26(10):2120–2123. [PMC free article] [PubMed]
  • Plikaytis BB, Plikaytis BD, Yakrus MA, Butler WR, Woodley CL, Silcox VA, Shinnick TM. Differentiation of slowly growing Mycobacterium species, including Mycobacterium tuberculosis, by gene amplification and restriction fragment length polymorphism analysis. J Clin Microbiol. 1992 Jul;30(7):1815–1822. [PMC free article] [PubMed]
  • Rogall T, Flohr T, Böttger EC. Differentiation of Mycobacterium species by direct sequencing of amplified DNA. J Gen Microbiol. 1990 Sep;136(9):1915–1920. [PubMed]
  • Shinnick TM. The 65-kilodalton antigen of Mycobacterium tuberculosis. J Bacteriol. 1987 Mar;169(3):1080–1088. [PMC free article] [PubMed]
  • Wayne LG, Sramek HA. Agents of newly recognized or infrequently encountered mycobacterial diseases. Clin Microbiol Rev. 1992 Jan;5(1):1–25. [PMC free article] [PubMed]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...