• 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. May 1997; 35(5): 1097–1100.
PMCID: PMC232709

Direct detection and identification of Mycobacterium ulcerans in clinical specimens by PCR and oligonucleotide-specific capture plate hybridization.

Abstract

We compared various diagnostic tests for their abilities to detect Mycobacterium ulcerans infection in specimens from patients with clinically active disease. Specimens from 10 patients from the area of Zangnanado (Department of Zou, Benin) with advanced, ulcerated active M. ulcerans infections were studied by direct smear, histopathology, culture, PCR, and oligonucleotide-specific capture plate hybridization (OSCPH). A total of 27 specimens, including 12 swabs of exudate collected before debridement and 15 fragments of tissue obtained during debridement, were submitted to bacteriologic and histopathologic analysis. The histopathologic evaluation of tissues from all six patients so tested revealed changes typical of those caused by M. ulcerans infection. Five specimens were contaminated, and M. ulcerans was cultivated on Löwenstein-Jensen medium from 12 of the remaining 22 (54.5%) specimens. Detection of mycobacteria was performed by PCR, and M. ulcerans was detected by OSCPH with a new probe (5'-CACGGGATTCATGTCCTGT-3') reacting with M. ulcerans and Mycobacterium marinum. In 10 of 22 (45.5%) specimens, M. ulcerans was identified by PCR-OSCPH. There was no statistically significant difference between the detection of M. ulcerans by culture and by PCR-OSCPH (P > 0.05). This is the first demonstration of an amplification system (PCR-OSCPH) with a sensitivity similar to that of culture for the direct and rapid recognition of M. ulcerans in clinical specimens. This system is capable of identifying M. ulcerans, even in paucibacillary lesions. Our findings suggest that PCR-OSCPH should be used in the quest for the elusive environmental reservoir(s) of M. ulcerans.

Full Text

The Full Text of this article is available as a PDF (124K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • De Beenhouwer H, Liang Z, De Rijk P, Van Eekeren C, Portaels F. Detection and identification of mycobacteria by DNA amplification and oligonucleotide-specific capture plate hybridization. J Clin Microbiol. 1995 Nov;33(11):2994–2998. [PMC free article] [PubMed]
  • Dumonceau JM, Fonteyne PA, Realini L, Van Gossum A, Van Vooren JP, Portaels F. Species-specific Mycobacterium genavense DNA in intestinal tissues of individuals not infected with human immunodeficiency virus. J Clin Microbiol. 1995 Sep;33(9):2514–2515. [PMC free article] [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]
  • Josse R, Guedenon A, Darie H, Anagonou S, Portaels F, Meyers WM. Les infections cutanées à Mycobacterium ulcerans: ulcères de Buruli. Med Trop (Mars) 1995;55(4):363–373. [PubMed]
  • Mangiapan G, Vokurka M, Schouls L, Cadranel J, Lecossier D, van Embden J, Hance AJ. Sequence capture-PCR improves detection of mycobacterial DNA in clinical specimens. J Clin Microbiol. 1996 May;34(5):1209–1215. [PMC free article] [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]
  • Meyers WM, Tignokpa N, Priuli GB, Portaels F. Mycobacterium ulcerans infection (Buruli ulcer): first reported patients in Togo. Br J Dermatol. 1996 Jun;134(6):1116–1121. [PubMed]
  • Pettit JH, Marchette NJ, Rees RJ. Mycobacterium ulcerans infection. Clinical and bacteriological study of the first cases recognized in South East Asia. Br J Dermatol. 1966 Apr;78(4):187–197. [PubMed]
  • Portaels F. Epidémiologie des ulcères à Mycobacterium ulcerans. Ann Soc Belg Med Trop. 1989 Jun;69(2):91–103. [PubMed]
  • Portaels F. Epidemiology of mycobacterial diseases. Clin Dermatol. 1995 May-Jun;13(3):207–222. [PubMed]
  • Portaels F, De Muynck A, Sylla MP. Selective isolation of mycobacteria from soil: a statistical analysis approach. J Gen Microbiol. 1988 Mar;134(3):849–855. [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]
  • Saxegaard F. Isolation of Mycobacterium paratuberculosis from intestinal mucosa and mesenteric lymph nodes of goats by use of selective Dubos medium. J Clin Microbiol. 1985 Aug;22(2):312–313. [PMC free article] [PubMed]
  • Lévy-Frébault VV, Portaels F. Proposed minimal standards for the genus Mycobacterium and for description of new slowly growing Mycobacterium species. Int J Syst Bacteriol. 1992 Apr;42(2):315–323. [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

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...