CULTURE OF THE CAUSATIVE ORGANISM OF DONOVANOSIS (CALYMMATOBACTERIUM-GRANULOMATIS) IN HEP-2 CELLS

We report successful culture of Calymmatobacterium granulomatis by sta ndard cell culture methods, Swabs were obtained from lesions in three patients with a clinical diagnosis of donovanosis, For two patients, t here was histological confirmation of the disease (i.e., the presence of Donovan bodies in Giemsa-stained smears), Specimens were inoculated onto cycloheximide-treated HEp-2 cell monolayers in RPMI 1640 medium (supplemented with fetal calf serum, NaHCO3, vancomycin hydrochloride, and benzylpenicillin). At 48 h, organisms resembling Donovan bodies w ere identified in monolayer cultures from all three specimens, The org anisms appeared as pleomorphic bacilli with characteristic bipolar sta ining and ''safety pin'' appearance, Using a PCR designed to different iate C. granulomatis from the Klebsiella species (which have a high de gree of molecular homology), we were able to demonstrate that the cult ured organisms produced a PCR product identical to that obtained from the original swab specimens, It is now possible to test in vitro susce ptibility of C. granulomatis to antibiotics and to provide a ready sou rce of DNA and antigenic material to enable the development of serolog ical tests and, possibly in the future, a vaccine.