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Journal of Clinical Microbiology, September 2003, p. 4231-4237, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4231-4237.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Development and Application of Real-Time PCR Assay for Quantification of Mycobacterium ulcerans DNA

S. Rondini,1* E. Mensah-Quainoo,2 H. Troll,3 T. Bodmer,4 and G. Pluschke1

Swiss Tropical Institute,1 Solvias AG, Basel,3 Institute for Infectious Diseases, Bern,Switzerland,4 Ghana Health Service, Amasaman, Ga District, Ghana2

Received 11 April 2003/ Returned for modification 6 June 2003/ Accepted 3 July 2003

Buruli ulcer, an infection caused by Mycobacterium ulcerans, is, after tuberculosis and leprosy, the third most common mycobacterial disease. The mode of transmission of M. ulcerans is not exactly known, but since Buruli ulcer often occurs in focalized swampy areas, it is assumed that there is a reservoir of the pathogen in stagnant water. Buruli ulcer usually starts as a painless nodule and can lead to massive destruction of skin, subcutaneous tissue, and eventually muscle and bone. Currently the only recommended treatment is wide surgical excision. In this report we describe the development of a real-time PCR method for the quantification of M. ulcerans DNA (IS2404 TaqMan). The highly specific assay is based on the detection of the M. ulcerans specific insertion sequence IS2404. The IS2404 TaqMan assay turned out to be about 10 times more sensitive than the available conventional PCR-based diagnostic test. It is demonstrated that the IS2404 TaqMan assay is suitable for the quantitative assessment of the dissemination of the mycobacteria in Buruli ulcer lesions. Prototype results obtained with excised tissue from a patient with a late preulcerative Buruli ulcer lesion reconfirmed earlier histopathological findings indicating that tissue damage occurs far beyond the regions in which large numbers of mycobacteria are detectable. The IS2404 TaqMan assay should be a useful tool for both diagnosis and research into the pathology and mode of transmission of this still inadequately investigated mycobacterial disease.

* Corresponding author. Mailing address: Socinst. 57, Swiss Tropical Institute, CH 4002 Basel, Switzerland. Phone: 41 61 2848277. Fax: 41 61 2718654. E-mail: Simona.Rondini{at}unibas.ch.

Journal of Clinical Microbiology, September 2003, p. 4231-4237, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4231-4237.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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