title: Dry reagent-based polymerase chain reaction compared with other laboratory methods available for the diagnosis of Buruli ulcer disease.
abstract: BACKGROUND
  Because of the multifaceted clinical presentation of Buruli ulcer disease, misclassification of clinically diagnosed cases may occur frequently. Laboratory tests for the confirmation of suspected cases include microscopic examination, culture, polymerase chain reaction (PCR), and histopathologic examination. However, microscopic examination, the only test usually available in areas of endemicity, has a low sensitivity.
  
  
  METHODS
  To make a highly sensitive diagnostic method locally available, dry reagent-based PCR (DRB-PCR), which is well adapted to tropical conditions, was pilot-tested in Ghana. Subsequently, the assay was used for the routine diagnosis of Buruli ulcer disease over a period of 2 years. The method was compared with other diagnostic tests to evaluate its performance under field conditions.
  
  
  RESULTS
  The interassay agreement rate between DRB-PCR and standard PCR was 91.7% for swab specimens and 95% for tissue specimens. Among all of the locally available tests, DRB-PCR revealed the highest overall positivity ratio. Sixty percent of patients with clinical diagnoses of Buruli ulcer disease had the diagnoses confirmed by DRB-PCR of swab or tissue specimens, compared with 30%-40% of patients who had diagnoses confirmed by microscopic examination of swab or tissue specimens. The positivity ratio of DRB-PCR varied considerably when analyzed per treatment center. Standardization of specimen collection resulted in a 30% increase in the positivity ratio of the assay, compared with that in the pilot-testing phase.
  
  
  CONCLUSIONS
  DRB-PCR is a reliable tool for the diagnosis of Buruli ulcer disease. However, PCR assays are suitable for detection only during early stages of the disease, when samples still contain bacilli. The quality of clinical diagnosis and the quality of diagnostic specimens strongly influence the positivity ratio.
authors.affiliation: Department of Infectious Diseases and Tropical Medicine, Ludwig-Maximilians University of Munich, Munich, Germany. vera.siegmund@lrz.uni-muenchen.de
authors.forename: Vera
authors.initials: V
authors.lastname: Siegmund
authors.forename: Ohene
authors.initials: O
authors.lastname: Adjei
authors.forename: Jörg
authors.initials: J
authors.lastname: Nitschke
authors.forename: William
authors.initials: W
authors.lastname: Thompson
authors.forename: Erasmus
authors.initials: E
authors.lastname: Klutse
authors.forename: Karl Heinz
authors.initials: KH
authors.lastname: Herbinger
authors.forename: Ruth
authors.initials: R
authors.lastname: Thompson
authors.forename: Felicitas
authors.initials: F
authors.lastname: van Vloten
authors.forename: Paul
authors.initials: P
authors.lastname: Racz
authors.forename: Bernhard
authors.initials: B
authors.lastname: Fleischer
authors.forename: Thomas
authors.initials: T
authors.lastname: Loescher
authors.forename: Gisela
authors.initials: G
authors.lastname: Bretzel
mesh_terms: D003937:Diagnosis, Differential; D005869:Ghana; D006801:Humans; D008828:Microbiological Techniques / Q000379:methods*; D009165:Mycobacterium Infections, Nontuberculous / Q000175:diagnosis* / Q000473:pathology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017192:Skin Diseases, Bacterial / Q000175:diagnosis* / Q000382:microbiology / Q000473:pathology