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Comparative Study
. 2012 Apr;50(4):1195-8.
doi: 10.1128/JCM.05592-11. Epub 2012 Jan 18.

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

Dissou Affolabi  1 , N'Dira Sanoussi, Koen Vandelannoote, Mathieu Odoun, Frank Faïhun, Ghislain Sopoh, Séverin Anagonou, Françoise Portaels, Miriam Eddyani
Affiliations
Comparative Study

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

Dissou Affolabi et al. J Clin Microbiol. 2012 Apr.

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.

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Figures

Fig 1
Fig 1
Summary of procedures used.
Fig 2
Fig 2
Distribution of specimens positive with at least one procedure before decontamination (n = 55).
Fig 3
Fig 3
Quantification of bacillary load by direct-smear examination versus quantitative real-time PCR for the 50 specimens positive by real-time PCR before decontamination. The values of all specimens are plotted with the average cycle threshold (CT) value per acid-fast bacillus (AFB).
Fig 4
Fig 4
Distribution of specimens positive with at least one procedure after decontamination (n = 46).
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