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Journal of Clinical Microbiology, January 2008, p. 69-72, Vol. 46, No. 1
0095-1137/08/$08.00+0     doi:10.1128/JCM.00301-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Primary Culture of Mycobacterium ulcerans from Human Tissue Specimens after Storage in Semisolid Transport Medium{triangledown}

Miriam Eddyani,1* Martine Debacker,1 Anandi Martin,1 Julia Aguiar,2 Christian R. Johnson,3 Cécile Uwizeye,1 Krista Fissette,1 and Françoise Portaels1

Mycobacteriology Unit, Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerpen, Belgium,1 Centre Sanitaire et Nutritionel Gbemoten, Zagnanado, Benin,2 Programme National de Lutte contre l'Ulcère de Buruli, Cotonou, Benin3

Received 7 February 2007/ Returned for modification 19 June 2007/ Accepted 30 September 2007

Tissue specimens collected from patients with clinically suspected Buruli ulcer treated in two Buruli ulcer treatment centers in Benin between 1998 and 2004 were placed in semisolid transport medium and transported at ambient temperature for microbiological analysis at the Institute of Tropical Medicine in Antwerp, Belgium. The impact of the delay before microbiological analysis on primary culture of Mycobacterium ulcerans was investigated. The length of storage in semisolid transport medium varied from 6 days to 26 weeks. Of the 1,273 tissue fragments positive for M. ulcerans DNA by an IS2404-specific PCR, 576 (45.2%) yielded positive culture results. The sensitivity of direct smear examination was 64.6% (822/1,273 tissue fragments). The median time required to obtain a positive culture result was 11 weeks. Positive cultures were obtained even from samples kept for more than 2 months at ambient temperatures. Moreover, there was no reduction in the viability of M. ulcerans, as detected by culture, when specimens remained in semisolid transport medium for long periods of time (up to 26 weeks). We can conclude that the method with semisolid transport medium is very robust for clinical specimens from patients with Buruli ulcer that, due to circumstances, cannot be analyzed in a timely manner. This transport medium is thus very useful for the confirmation of a diagnosis of Buruli ulcer with specimens collected in the field.

* Corresponding author. Mailing address: Mycobacteriology Unit, Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerpen, Belgium. Phone: 32-32476548. Fax: 32-32476333. E-mail: meddyani{at}itg.be

{triangledown} Published ahead of print on 7 November 2007.

Journal of Clinical Microbiology, January 2008, p. 69-72, Vol. 46, No. 1
0095-1137/08/$08.00+0     doi:10.1128/JCM.00301-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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