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

D Affolabi, ND Sanoussi, K Vandelannoote, M Odoun, F Faihun, G Sopoh, S Anagonou, F Portaels, M Eddyani

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Abstract

We compared two DNA extraction methods (a semi-automated method using Maxwell(R) 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 clinically suspected Buruli ulcer patients. All 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(R) 16 extraction method, performed on non-decontaminated suspensions are the best for the molecular diagnosis of Mycobacterium ulcerans disease.
Original languageEnglish
JournalJournal of Clinical Microbiology
Volume50
Issue number4
Pages (from-to)1195-1198
ISSN0095-1137
DOIs
Publication statusPublished - 2012

Keywords

  • B780-tropical-medicine
  • Bacterial diseases
  • Buruli ulcer
  • Mycobacterium ulcerans
  • Molecular diagnostic techniques
  • Decontamination
  • DNA extraction
  • Amplification
  • Polymerase chain reaction
  • PCR
  • Comparison
  • Methodology
  • Smear-positive
  • Laboratory techniques and procedures

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