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J. Clin. Microbiol. doi:10.1128/JCM.00301-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

PRIMARY CULTURE OF MYCOBACTERIUM ULCERANS FROM HUMAN TISSUE SPECIMENS AFTER STORAGE IN A SEMI-SOLID TRANSPORT MEDIUM

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

	Abstract

Tissue specimens collected from clinically suspected Buruli ulcer patients treated in two Buruli ulcer treatment centres in Benin between 1998 and 2004 were placed in a semi-solid transport medium and transported at ambient temperature for microbiological analysis in the Institute of Tropical Medicine in Antwerp, Belgium. The impact of the delay before microbiological analysis on primary culture of M. ulcerans was investigated. Storage in semi-solid transport medium varied between 6 days up to 26 weeks.

Among the 1273 tissue fragments positive for M. ulcerans-DNA by IS2404-PCR, 576 (45.2%) yielded cultures. The sensitivity of direct smear examination was 64.6% (822/1273). The median time to obtain a positive culture was 11 weeks. Positive cultures were obtained even from samples kept more than two months at ambient temperatures. Moreover, there was no reduction in viability of M. ulcerans as detected by culture when specimens remain for long periods of time (up to 26 weeks) in a semi-solid transport medium.

We can conclude that the semi-solid transport method used is very robust for clinical specimens from patients with Buruli ulcer that, due to circumstances, can not be timely analysed. This transport medium is thus very useful for the confirmation of diagnosis of Buruli ulcer in specimens collected in the field.