Seven different agar-based media were compared to determine the optimal set of culture media for primary isolation of . Also, a new method for sampling genital ulcers – with a disposable sterile plastic loop – and processing specimens that provides a standardised inoculum for culture of on various media is described. A total of 202 patients with genital ulcer disease was enrolled in this study. A sterile swab or plastic loop was used to sample the base of the ulcers and ulcer material was suspended in sterile phosphate-buffered saline. A 100-μl sample of this suspension was mixed with an equal volume of tryptic soy broth containing IsoVitaleX and centrifuged for 1 min. This suspension was used to inoculate the different media. Plates were incubated at 33°C in micro-aerophilic conditions and examined for growth of after 48 h. Of the 202 specimens, 77 (38.1%) were culture positive for . None of the agar bases supported the growth of all strains. Based on this observation, we recommend the universal use of Mueller-Hinton agar base supplemented with chocolate horse blood and IsovitaleX (MH-HBC) and Columbia agar base supplemented with bovine haemoglobin, activated charcoal, fetal calf serum and IsovitaleX (C-HgCh) to enable comparison of prevalence figures for chancroid. In addition, the novel sampling technique described in this study eliminates sampling bias normally associated with genital ulcer specimens.


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