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A comparison of DNA extraction procedures for the detection of Mycobacterium ulcerans, the causative agent of Buruli ulcer, in clinical and environmental specimens

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dc.contributor.author Durnez, L.
dc.contributor.author Stragier, P.
dc.contributor.author Roebben, K.
dc.contributor.author Ablordey, A.
dc.contributor.author Leirs, H.
dc.contributor.author Portaels, F.
dc.date.accessioned 2009-02-13T10:39:38Z
dc.date.available 2009-02-13T10:39:38Z
dc.date.issued 2009
dc.identifier.issn 0167-7012
dc.identifier.doi http://dx.doi.org/10.1016/j.mimet.2008.10.002
dc.identifier.other ITG-M1B
dc.identifier.other ITG-M2B
dc.identifier.other ITG-M3B
dc.identifier.other ITG-M4B
dc.identifier.other ITG-MLA
dc.identifier.other MICRO
dc.identifier.other U-MYCOB
dc.identifier.other JIF
dc.identifier.other DOI
dc.identifier.other UPD9
dc.identifier.other ABSTRACT
dc.identifier.uri http://hdl.handle.net/10390/2509
dc.description.abstract Mycobacterium ulcerans is the causative agent of Buruli ulcer, the third most common mycobacterial disease in humans after tuberculosis and leprosy. Although the disease is associated with aquatic ecosystems, cultivation of the bacillus from the environment is difficult to achieve. Therefore, at the moment, research is based on the detection by PCR of the insertion sequence IS2404 present in M. ulcerans and some closely related mycobacteria. In the present study, we compared four DNA extraction methods for detection of M. ulcerans DNA, namely the one tube cell lysis and DNA extraction procedure (OT), the FastPrep procedure (FP), the modified Boom procedure (MB), and the Maxwell® 16 Procedure (M16). The methods were performed on serial dilutions of M. ulcerans, followed by PCR analysis with different PCR targets in M. ulcerans to determine the detection limit (DL) of each method. The purity of the extracted DNA and the time and effort needed were compared as well. All methods were performed on environmental specimens and the two best methods (MB and M16) were tested on clinical specimens for detection of M. ulcerans DNA. When comparing the DLs of the DNA extraction methods, the MB and M16 had a significantly lower DL than the OT and FP. For the different PCR targets, IS2404 showed a significantly lower DL than mlsA, MIRU1, MIRU5 and VNTR6. The FP and M16 were considerably faster than the MB and OT, while the purity of the DNA extracted with the MB was significantly higher than the DNA extracted with the other methods. The MB performed best on the environmental and clinical specimens. This comparative study shows that the modified Boom procedure, although lengthy, provides a better method of DNA extraction than the other methods tested for detection and identification of M. ulcerans in both clinical and environmental specimens.
dc.language English en
dc.subject Bacteriology en
dc.subject Mycobacterium ulcerans en
dc.subject DNA extraction en
dc.subject Procedures en
dc.subject Comparative study en
dc.title A comparison of DNA extraction procedures for the detection of Mycobacterium ulcerans, the causative agent of Buruli ulcer, in clinical and environmental specimens en
dc.type Article en
dc.citation.issue 2 en
dc.citation.jtitle Journal of Microbiological Methods en
dc.citation.volume 76 en
dc.citation.pages 152-158 en
dc.identifier.pmid http://www.ncbi.nlm.nih.gov/pubmed/18973778
dc.citation.jabbreviation J Microbiol Meth en


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