02853nas a2200349   4500000000100000008004100001260001300042653001200055653001700067653003000084653001900114653001500133653002900148653001100177653002700188653003000215653002900245653003200274653002200306653002300328100001300351700001500364700001400379700001500393700001200408700001500420245016900435300001000604490000700614520186800621022001402489       2009                            d  c2009 Feb10aAnimals10aBuruli ulcer10aDNA Transposable Elements10aDNA, Bacterial10aDNA, Plant10aEnvironmental Monitoring10aHumans10aMycobacterium ulcerans10apolymerase chain reaction10aReagent Kits, Diagnostic10aSensitivity and Specificity10aSoil Microbiology10aWater Microbiology1 aDurnez L1 aStragier P1 aRoebben K1 aAblordey A1 aLeirs H1 aPortaels F00aA comparison of DNA extraction procedures for the detection of Mycobacterium ulcerans, the causative agent of Buruli ulcer, in clinical and environmental specimens.  a152-80 v763 a<p>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.</p>  a0167-7012