Phylogenetic analysis of Calymmatobacterium granulomatis based on 16S rRNA gene sequences.
Calymmatobacterium granulomatis is the aetiological agent of granuloma inguinale - a chronic granulomatous genital infection - and is morphologically similar to members of the genus Klebsiella. This study determined the 16S rRNA gene sequence of C. granulomatis and the taxonomic position of the organism in relation to the genus Klebsiella. Genomic DNA was extracted from C. granulomatis-infected monocytes and from frozen and formalin-fixed paraffin wax-embedded tissue biopsy specimens from patients with histologically proven granuloma inguinale. The 16S rDNA was amplified by PCR with broad range oligonucleotide primers. The amplified DNA fragments were cloned into pMOS vector, digested with Bam HI and Pst1 restriction endonucleases, hybridised with a gram-negative bacterial probe (DL04), sequenced in both directions by the automated ALF DNA sequencer, verified on an ABI Prism 377 automated sequencer and analysed with DNASIS and MEGA software packages. Sequence analysis revealed DNA homology of 99% in C. granulomatis from the different sources, supporting the belief that the bacteria in the culture and the biopsy specimens belonged to the same species, although there was some diversity within the species. Phylogenetically, the strains were closely related to the genera Klebsiella and Enterobacter with similarities of 95% and 94% respectively. C. granulomatis is a unique species, distinct from other related organisms belonging to the gamma subclass of Proteobacteria. (+info)
Phylogenetic evidence for reclassification of Calymmatobacterium granulomatis as Klebsiella granulomatis comb. nov.
By sequencing a total of 2089 bp of the 16S rRNA and phoE genes it was demonstrated that Calymmatobacterium granulomatis (the causative organism of donovanosis) shows a high level of identity with Klebsiella species pathogenic to humans (Klebsiella pneumoniae, Klebsiella rhinoscleromatis). It is proposed that C. granulomatis should be reclassified as Klebsiella granulomatis comb. nov. An emended description of the genus Klebsiella is given. (+info)
A colorimetric detection system for Calymmatobacterium granulomatis.
OBJECTIVE: To incorporate the first polymerase chain reaction (PCR) assay for Calymmatobacterium granulomatis into a colorimetric detection system for use in routine diagnostic laboratories. METHODS: A capture oligonucleotide specific for the Klebsiella phoE gene was covalently linked to tosyl activated magnetic beads. Biotinylated phoE PCR products obtained from 14 positive specimens from patients with donovanosis and isolates of Klebsiella pneumoniae, K rhinoscleromatis, and K ozaenae were cleaved with HaeIII for the purpose of differentiation, captured by the prepared beads, and subjected to standard EIA detection methodology. Eight samples from unrelated genital conditions underwent the same procedure. It was anticipated from the sequence data that the biotinylated fragment would be cleaved from the capture oligonucleotide target region in the three Klebsiella phoE products (that is, a negative colorimetric result) while the entire fragment of interest would remain intact in the positive C granulomatis phoE products (that is, a positive colorimetric result). RESULTS: All 14 positive specimens from patients with donovanosis gave strong colorimetric readings with this detection system. Isolates of K pneumoniae, K rhinoscleromatis, K ozaenae, and the eight specimens from unrelated genital conditions were negative. CONCLUSION: The successful development of a colorimetric detection system for C granulomatis incorporating two levels of specificity enables the molecular diagnosis of this condition to be undertaken by routine diagnostic laboratories. This should have an important role in the Australian government's campaign to eradicate donovanosis by 2003 though the test still needs to undergo trials and be validated using a larger number of samples from geographically diverse parts of the world in order to ascertain the generalisability of the methodology. (+info)
Dorsal perforation of prepuce: a common end point of severe ulcerative genital diseases?
Severe ulcerative genital diseases can cause destruction of the prepuce, glans, or sometimes of the whole penis (phagedena). We observed a characteristic pattern of partial destruction of the prepuce as a result of a wide variety of ulcerative genital diseases. Five patients, two with severe genital herpes, one with hidradenitis suppurativa, and two with donovanosis presented with perforation on the dorsal surface of the prepuce. In four of them, the glans protruded through the defect and in one, the defect was not large enough to allow protrusion of the glans. In two patients, the preputial sac was obliterated. The relatively decreased blood supply of the prepuce is the probable explanation for perforation at this selective site. (+info)
Simplified microimmunofluorescence test with trachoma-lymphogranuloma venereum (Chlamydia trachomatis) antigens for use as a screening test for antibody.
A simplified microimmunofluorescence test with trachoma-lymphogranuloma venereum (Chlamydia trachomatis) antigens has been devised as a screening test for antibody in human sera. The test differs from our standard procedure by amalgamating 15 different immunotypes into nine antigen pools, by using only three serum dilutions, and by dropping use of duplicate slides. The screening test could be performed on at least six times as many sera as the standard test for a given unit of effort. It was shown to have a sensitivity equal to the standard test and an ability to determine specific immunotype on the basis of antibody pattern (72%) only slightly reduced from the standard test (84%). The screening test was carried out on 876 patients from different population groups in the Seattle area. The tests showed that antibody was present frequently in persons attending venereal disease clinics (60%) and commonly (25%) in a group of adults without venereal disease. Nine percent of children under 15 years of age tested had antibody; the significance of this finding is unknown. (+info)
Donovanosis, a chronic cause of genital ulceration, has recently been the subject of renewed interest after a long period of relative obscurity. The causative organism, Calymmatobacterium granulomatis, has been cultured for the first time in many years and a polymerase chain reaction diagnostic using a colorimetric detection system has been developed. Phylogenetic analysis confirms close similarities with the genus Klebsiella and a proposal made that C granulomatis be reclassified as Klebsiella granulomatis comb nov. Azithromycin has emerged as the drug of choice and should be used if the diagnosis is confirmed or suspected. In donovanosis endemic areas, syndromic management protocols for genital ulceration may need to be adapted locally. A significant donovanosis epidemic was reported in Durban from 1988-97 but the current status of this epidemic is unclear. The donovanosis elimination programme among Aboriginals in Australia appears successful and is a model that could be adopted in other donovanosis endemic areas. Overall, the incidence of donovanosis seems to be decreasing. Increased attention would undoubtedly be paid to donovanosis if policy makers recognised more readily the importance of genital ulcers in fuelling the HIV epidemic. (+info)
A serological test for granuloma inguinale.
OBJECTIVES: An indirect immunofluorescence technique applied to paraffin embedded tissue sections of lesions containing Donovan bodies was evaluated as a serological test for the diagnosis of granuloma inguinale. METHODS: Sera from patients with proven granuloma inguinale, other sexually acquired genital ulcerations and blood donors from areas where granuloma inguinale is rarely encountered as well as from disease-endemic regions were tested. Sera were tested either unabsorbed or following absorption with whole Klebsiella pneumoniae bacteria. RESULTS: Using unabsorbed sera at a dilution of 1:160 the test was found to have a sensitivity of 100%, specificity of 98%, positive predictive value (PPV) of 89% and negative predictive value (NPV) of 100%. There proved to be no advantage in preabsorbing sera with K. pneumoniae antigen. CONCLUSIONS: In the absence of culture methods for Calymmatobacterium granulomatis, an indirect immunofluorescence technique may prove valuable for the diagnosis of individual cases of granuloma inguinale and as an epidemiological tool in studies of the disease. (+info)
Trends in reported cases of donovanosis in Durban, South Africa.
OBJECTIVE: To investigate recent trends in reported cases of donovanosis (granuloma inguinale) in Durban, South Africa. DESIGN: The annual reports of the Medical Officer of Health for Durban 1958-1988 were reviewed to identify cases of donovanosis, genital ulcer disease (GUD) and new patients with sexually transmitted diseases (STD). A rapid staining technique for the detection of Donovan bodies was introduced in 1988. SETTING: City Health STD Clinic, King Edward VIII Hospital, Durban. RESULTS: An initial peak was identified in men 1969-1974. A second peak was recorded in 1988 when reported cases of donovanosis (313) were the highest since records commenced. Both peaks were unrelated to either increases in the numbers of new attenders with STD or patients with GUD. CONCLUSION: The recent increase in donovanosis in Durban may reflect either a new epidemic or under-reporting of a disease previously diagnosed on clinical grounds. Improved control of donovanosis, a condition sometimes causing extensive GUD, and which has been implicated in HIV-1 transmission in local men, should be targeted in HIV control programmes. (+info)