• 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. Jul 1997; 35(7): 1696–1700.
PMCID: PMC229824

Development of a PCR assay for rapid diagnosis of Mycobacterium ulcerans infection.

Abstract

The diagnosis of Mycobacterium ulcerans infection is hampered by the slow growth of the bacterium in culture, resulting in a delay of several months before a specific diagnosis can be obtained. In addition, M. ulcerans cannot be isolated from water even when there is convincing epidemiological evidence implicating this as the source of infection. The aim of the present study was to develop a PCR assay to circumvent the problems of delayed diagnosis and insensitivity of standard bacterial culture for M. ulcerans. For the PCR, we isolated an M. ulcerans-specific DNA fragment, 1,109 bp long, which is repeated at least 50 times throughout the genome. Use of this sequence as a target for PCR allowed us to detect as few as 2 molecules of genomic DNA in vitro. The PCR was used to detect M. ulcerans DNA in fresh tissue and paraffin-embedded sections from all seven patients with culture-confirmed cases of infection.

Full Text

The Full Text of this article is available as a PDF (1.0M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Beige J, Lokies J, Schaberg T, Finckh U, Fischer M, Mauch H, Lode H, Köhler B, Rolfs A. Clinical evaluation of a Mycobacterium tuberculosis PCR assay. J Clin Microbiol. 1995 Jan;33(1):90–95. [PMC free article] [PubMed]
  • Buchanan TM, Nomaguchi H, Anderson DC, Young RA, Gillis TP, Britton WJ, Ivanyi J, Kolk AH, Closs O, Bloom BR, et al. Characterization of antibody-reactive epitopes on the 65-kilodalton protein of Mycobacterium leprae. Infect Immun. 1987 Apr;55(4):1000–1003. [PMC free article] [PubMed]
  • Collins DM, Stephens DM. Identification of an insertion sequence, IS1081, in Mycobacterium bovis. FEMS Microbiol Lett. 1991 Sep 15;67(1):11–15. [PubMed]
  • Flood P, Street A, O'Brien P, Hayman J. Mycobacterium ulcerans infection on Phillip Island, Victoria. Med J Aust. 1994 Feb 7;160(3):160–160. [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]
  • Hayman J. Clinical features of Mycobacterium ulcerans infection. Australas J Dermatol. 1985 Aug;26(2):67–73. [PubMed]
  • Hayman J. Postulated epidemiology of Mycobacterium ulcerans infection. Int J Epidemiol. 1991 Dec;20(4):1093–1098. [PubMed]
  • Hayman J, McQueen A. The pathology of Mycobacterium ulcerans infection. Pathology. 1985 Oct;17(4):594–600. [PubMed]
  • Hernandez Perez M, Fomukong NG, Hellyer T, Brown IN, Dale JW. Characterization of IS1110, a highly mobile genetic element from Mycobacterium avium. Mol Microbiol. 1994 Jun;12(5):717–724. [PubMed]
  • Hofer M, Hirschel B, Kirschner P, Beghetti M, Kaelin A, Siegrist CA, Suter S, Teske A, Böttger EC. Brief report: disseminated osteomyelitis from Mycobacterium ulcerans after a snakebite. N Engl J Med. 1993 Apr 8;328(14):1007–1009. [PubMed]
  • Johnson PD, Veitch MG, Flood PE, Hayman JA. Mycobacterium ulcerans infection on Phillip Island, Victoria. Med J Aust. 1995 Feb 20;162(4):221–222. [PubMed]
  • Johnson PD, Veitch MG, Leslie DE, Flood PE, Hayman JA. The emergence of Mycobacterium ulcerans infection near Melbourne. Med J Aust. 1996 Jan 15;164(2):76–78. [PubMed]
  • Kunze ZM, Wall S, Appelberg R, Silva MT, Portaels F, McFadden JJ. IS901, a new member of a widespread class of atypical insertion sequences, is associated with pathogenicity in Mycobacterium avium. Mol Microbiol. 1991 Sep;5(9):2265–2272. [PubMed]
  • LUNN HF, CONNOR DH, WILKS NE, BARNLEY GR, KAMUNVI F, CLANCEY JK, BEE JD. BURULI (MYCOBACTERIAL) ULCERATION IN UGANDA. (A NEW FOCUS OF BURULI ULCER IN MADI DISTRICT, UGANDA): REPORT OF A FIELD STUDY. East Afr Med J. 1965 Jun;42:275–288. [PubMed]
  • Marston BJ, Diallo MO, Horsburgh CR, Jr, Diomande I, Saki MZ, Kanga JM, Patrice G, Lipman HB, Ostroff SM, Good RC. Emergence of Buruli ulcer disease in the Daloa region of Cote d'Ivoire. Am J Trop Med Hyg. 1995 Mar;52(3):219–224. [PubMed]
  • Portaels F. Epidémiologie des ulcères à Mycobacterium ulcerans. Ann Soc Belg Med Trop. 1989 Jun;69(2):91–103. [PubMed]
  • Portaels F, Fonteyene PA, de Beenhouwer H, de Rijk P, Guédénon A, Hayman J, Meyers MW. Variability in 3' end of 16S rRNA sequence of Mycobacterium ulcerans is related to geographic origin of isolates. J Clin Microbiol. 1996 Apr;34(4):962–965. [PMC free article] [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]
  • Thierry D, Brisson-Noël A, Vincent-Lévy-Frébault V, Nguyen S, Guesdon JL, Gicquel B. Characterization of a Mycobacterium tuberculosis insertion sequence, IS6110, and its application in diagnosis. J Clin Microbiol. 1990 Dec;28(12):2668–2673. [PMC free article] [PubMed]
  • Tsang AY, Farber ER. The primary isolation of mycobacterium ulcerans. Am J Clin Pathol. 1973 May;59(5):688–692. [PubMed]
  • Vary PH, Andersen PR, Green E, Hermon-Taylor J, McFadden JJ. Use of highly specific DNA probes and the polymerase chain reaction to detect Mycobacterium paratuberculosis in Johne's disease. J Clin Microbiol. 1990 May;28(5):933–937. [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