title: 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.
authors.affiliation: Laboratoire de Référence des Mycobactéries, Cotonou, Benin.
authors.forename: Dissou
authors.initials: D
authors.lastname: Affolabi
authors.forename: N'Dira
authors.initials: N
authors.lastname: Sanoussi
authors.forename: Koen
authors.initials: K
authors.lastname: Vandelannoote
authors.forename: Mathieu
authors.initials: M
authors.lastname: Odoun
authors.forename: Frank
authors.initials: F
authors.lastname: Faïhun
authors.forename: Ghislain
authors.initials: G
authors.lastname: Sopoh
authors.forename: Séverin
authors.initials: S
authors.lastname: Anagonou
authors.forename: Françoise
authors.initials: F
authors.lastname: Portaels
authors.forename: Miriam
authors.initials: M
authors.lastname: Eddyani
mesh_terms: D058491:Bacterial Load; D054312:Buruli Ulcer / Q000175:diagnosis* / Q000382:microbiology; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D003666:Decontamination; D005798:Genes, Bacterial; D006801:Humans; D025202:Molecular Diagnostic Techniques; D019911:Mycobacterium ulcerans / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial