title: Analysis of an IS2404-based nested PCR for diagnosis of Buruli ulcer disease in regions of Ghana where the disease is endemic.
abstract: Mycobacterium ulcerans causes Buruli ulcer disease (BUD), an ulcerative skin disease emerging mainly in West Africa. Laboratory confirmation of BUD is complicated as no "gold standard" for diagnosis exists. A nested primer PCR based on IS2404 has shown promise as a diagnostic assay. We evaluated the IS2404-based PCR to detect M. ulcerans DNA in tissue specimens from 143 BUD patients diagnosed according to the World Health Organization BUD clinical case definition in Ghana. Comparisons were made with culture and histopathology results. Variables influencing detection rate tested in this PCR protocol included the amount of tissue used and the stage of disease. The nested PCR was repeated on DNA extracted from a different part of the same biopsy specimen of 21 culture-positive samples. Of all 143 specimens, 107 (74.8%; 95% confidence interval, 68 to 82%) showed the presence of M. ulcerans DNA by PCR. Of the 78 histology-confirmed BUD patient samples, 64 (83%) were PCR positive. Detection rates were influenced neither by the amount of tissue processed for PCR nor by the stage of disease (preulcerative or ulcerative). Taken together, the two nested PCR tests on the subset of 21 culture-positive samples were able to detect M. ulcerans DNA in all 21 culture-confirmed patients. For future studies, small tissue samples, e.g., punch biopsy samples, might be sufficient for case confirmation.
authors.affiliation: Department of Medicine, Emory University School of Medicine, Atlanta, GA 30303, USA.
authors.forename: Ymkje
authors.initials: Y
authors.lastname: Stienstra
authors.forename: Tjip S
authors.initials: TS
authors.lastname: van der Werf
authors.forename: Jeannette
authors.initials: J
authors.lastname: Guarner
authors.forename: Pratima L
authors.initials: PL
authors.lastname: Raghunathan
authors.forename: Ellen A
authors.initials: EA
authors.lastname: Spotts Whitney
authors.forename: Winette T A
authors.initials: WT
authors.lastname: van der Graaf
authors.forename: Kwame
authors.initials: K
authors.lastname: Asamoa
authors.forename: Jordan W
authors.initials: JW
authors.lastname: Tappero
authors.forename: David A
authors.initials: DA
authors.lastname: Ashford
authors.forename: C Harold
authors.initials: CH
authors.lastname: King
mesh_terms: D016022:Case-Control Studies; D004251:DNA Transposable Elements; D019353:Endemic Diseases; D005869:Ghana / Q000453:epidemiology; D006801:Humans; D009165:Mycobacterium Infections, Nontuberculous / Q000453:epidemiology* / Q000382:microbiology; D019911:Mycobacterium ulcerans / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012883:Skin Ulcer / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology
chemical_list: D004251:DNA Transposable Elements