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Effects of decontamination, DNA extraction, and amplification procedures on the molecular diagnosis of Mycobacterium ulcerans disease (Buruli ulcer).

Abstract
We compared two DNA extraction methods (a semiautomated method using a Maxwell kit and a modified Boom method) and three amplification procedures (a single-step PCR, a nested PCR, and a real-time quantitative PCR) on 74 surgical tissue specimens from patients with clinically suspected Buruli ulcer. All of these procedures were compared before and after decontamination. We observed that, among the procedures tested, real-time PCR after the modified Boom extraction method or a single-run PCR assay after the Maxwell 16 extraction method, performed on nondecontaminated suspensions, are the best for the molecular diagnosis of Mycobacterium ulcerans disease.
AuthorsDissou Affolabi, N'Dira Sanoussi, Koen Vandelannoote, Mathieu Odoun, Frank Faïhun, Ghislain Sopoh, Séverin Anagonou, Françoise Portaels, Miriam Eddyani
JournalJournal of clinical microbiology (J Clin Microbiol) Vol. 50 Issue 4 Pg. 1195-8 (Apr 2012) ISSN: 1098-660X [Electronic] United States
PMID22259213 (Publication Type: Comparative Study, Journal Article, Multicenter Study, Research Support, Non-U.S. Gov't)
Chemical References
  • DNA Transposable Elements
  • DNA, Bacterial
Topics
  • Bacterial Load
  • Buruli Ulcer (diagnosis, microbiology)
  • DNA Transposable Elements (genetics)
  • DNA, Bacterial (genetics, isolation & purification)
  • Decontamination
  • Genes, Bacterial
  • Humans
  • Molecular Diagnostic Techniques
  • Mycobacterium ulcerans (genetics)
  • Real-Time Polymerase Chain Reaction

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