Primary culture of Mycobacterium ulcerans from human tissue specimens after storage in semisolid transport medium.
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.
Eddyani
Miriam
M
Mycobacteriology Unit, Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerpen, Belgium. meddyani@itg.be
Debacker
Martine
M
Martin
Anandi
A
Aguiar
Julia
J
Johnson
Christian R
CR
Uwizeye
Cécile
C
Fissette
Krista
K
Portaels
Françoise
F
D001431:Bacteriological Techniques / Q000379:methods*; D001530:Belgium; D001541:Benin; D054312:Buruli Ulcer / Q000175:diagnosis / Q000382:microbiology; D015169:Colony Count, Microbial; D003470:Culture Media / Q000737:chemistry*; D006801:Humans; D050296:Microbial Viability; D019911:Mycobacterium ulcerans / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*; D013997:Time Factors
D003470:Culture Media