@article {13, title = {A comparison of DNA extraction procedures for the detection of Mycobacterium ulcerans, the causative agent of Buruli ulcer, in clinical and environmental specimens.}, journal = {J Microbiol Methods}, volume = {76}, year = {2009}, month = {2009 Feb}, pages = {152-8}, 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.

}, keywords = {Animals, Buruli Ulcer, DNA Transposable Elements, DNA, Bacterial, DNA, Plant, Environmental Monitoring, Humans, Mycobacterium ulcerans, Polymerase Chain Reaction, Reagent Kits, Diagnostic, Sensitivity and Specificity, Soil Microbiology, Water Microbiology}, issn = {0167-7012}, doi = {10.1016/j.mimet.2008.10.002}, author = {Durnez, Lies and Stragier, Pieter and Roebben, Karen and Ablordey, Anthony and Leirs, Herwig and Portaels, Fran{\c c}oise} } @article {14, title = {Persistence of Mycobacterium ulcerans disease (Buruli Ulcer) in the historical focus of Kasongo Territory, the Democratic Republic of Congo.}, journal = {Am J Trop Med Hyg}, volume = {81}, year = {2009}, month = {2009 Nov}, pages = {888-94}, abstract = {

Fifty years after the last report of Mycobacterium ulcerans infections (Buruli ulcer [BU]) in Kasongo Territory, Maniema Province, Democratic Republic of Congo (DRC), we conducted a small-scale cross-sectional survey to assess if this historical BU focus was still active and if so to explore the disease epidemiology. Seventy-five active and inactive BU cases were identified on clinical grounds of which two of 28 BU active cases were laboratory confirmed. We used a modified BU02 form to reconstruct the local disease dynamics and we believe that the horrific conflict in eastern DRC and exceptional flooding were the most likely causes of the re-emergence of the disease. There is a need in the DRC to decentralize and integrate surveillance and control activities at local level to increase the effectiveness of patient management.

}, keywords = {Adolescent, Adult, Age Distribution, Aged, Buruli Ulcer, Child, Democratic Republic of the Congo, Female, Humans, Male, Middle Aged, Mycobacterium ulcerans, Sex Characteristics, Young Adult}, issn = {1476-1645}, doi = {10.4269/ajtmh.2009.09-0049}, author = {Suykerbuyk, Patrick and Wambacq, Julie and Phanzu, Delphin M and Haruna, Hemedi and Nakazawa, Yoshinori and Ooms, Kristien and Kamango, Kalambo and Stragier, Pieter and Singa, Jackie N and Ekwanzala, Florent and De Herdt, Eric and De Maeyer, Philippe and Kestens, Luc and Portaels, Fran{\c c}oise} } @article {15, title = {Buruli ulcer in United Kingdom tourist returning from Latin America.}, journal = {Emerg Infect Dis}, volume = {15}, year = {2009}, month = {2009 Nov}, pages = {1827-9}, abstract = {

We report a case of Buruli ulcer in a tourist from the United Kingdom. The disease was almost certainly acquired in Brazil, where only 1 case had previously been reported. The delay in diagnosis highlights the need for physicians to be aware of the disease and its epidemiology.

}, keywords = {Adult, Anti-Bacterial Agents, Brazil, Buruli Ulcer, Communicable Diseases, Emerging, Great Britain, Humans, Latin America, Male, Mycobacterium ulcerans, Skin Transplantation, Travel}, issn = {1080-6059}, doi = {10.3201/eid1511.090460}, author = {McGann, Hugh and Stragier, Pieter and Portaels, Fran{\c c}oise and Gascoyne Binzi, Deborah and Collyns, Timothy and Lucas, Sebastian and Mawer, Damian} } @article {16, title = {Usefulness of the European Epidemic Intelligence Information System in the management of an outbreak of listeriosis, Belgium, 2011.}, journal = {Euro Surveill}, volume = {17}, year = {2012}, month = {2012}, abstract = {

A cluster of time-linked cases and the identification of a clonal strain suggest the occurrence of an outbreak of listeriosis in Belgium in 2011, presumably due to the consumption of hard cheese made with pasteurised milk and produced by a Belgium manufacturer. The outbreak clone was identified as Listeria monocytogenes serovar 1/2a, sensitive to arsenic and cadmium and of multilocus sequence typing MLST-type 37. Food investigation of this outbreak was facilitated by the European Epidemic Intelligence Information System and data exchanged between French and Belgium listeriosis surveillance systems.

}, keywords = {Aged, Aged, 80 and over, Arsenites, Bacterial Typing Techniques, Belgium, Cadmium Chloride, Cluster Analysis, Disease Outbreaks, Drug Resistance, Microbial, Electrophoresis, Gel, Pulsed-Field, Europe, Female, Foodborne Diseases, Geographic Information Systems, Hospitalization, Humans, Information Dissemination, Listeria monocytogenes, Listeriosis, Male, Microbial Sensitivity Tests, Middle Aged, Molecular Sequence Data, Population Surveillance}, issn = {1560-7917}, author = {Yde, M and Naranjo, M and Mattheus, W and Stragier, P and Pochet, B and Beulens, K and De Schrijver, K and Van den Branden, D and Laisnez, V and Flipse, W and Leclercq, A and Lecuit, M and Dierick, K and Bertrand, S} } @article {3, title = {Comparative nucleotide sequence analysis of polymorphic variable-number tandem-repeat Loci in Mycobacterium ulcerans.}, journal = {J Clin Microbiol}, volume = {43}, year = {2005}, month = {2005 Oct}, pages = {5281-4}, abstract = {

We analyzed a set of variable-number tandem-repeat (VNTR) loci to assess their nucleotide sequence diversity in isolates of three Mycobacterium ulcerans genotypes. Sequence variants in two loci resulted in intraspecies resolution of Southeast Asian and Asian genotypes in contrast to a homogenous sequence composition among African isolates. Nucleotide sequence polymorphism in repeat units can enhance discrimination of VNTR loci.

}, keywords = {Bacterial Typing Techniques, Base Sequence, Genetic Variation, Genotype, Humans, Minisatellite Repeats, Molecular Sequence Data, Mycobacterium ulcerans, Polymorphism, Genetic, Sequence Alignment, Species Specificity}, issn = {0095-1137}, doi = {10.1128/JCM.43.10.5281-5284.2005}, author = {Ablordey, Anthony and Hilty, Markus and Stragier, Pieter and Swings, Jean and Portaels, Fran{\c c}oise} } @article {5, title = {Heterogeneity among Mycobacterium ulcerans isolates from Africa.}, journal = {Emerg Infect Dis}, volume = {12}, year = {2006}, month = {2006 May}, pages = {844-7}, abstract = {

Mycobacterium ulcerans causes Buruli ulcer, an ulcerative skin disease in tropical and subtropical areas. Despite restricted genetic diversity, mycobacterial interspersed repetitive unit-variable-number tandem repeat analysis on M. ulcerans revealed 3 genotypes from different African countries. It is the first time this typing method succeeded directly on patient samples.

}, keywords = {Africa, Bacterial Typing Techniques, Base Sequence, Genetic Variation, Genotype, Humans, Molecular Epidemiology, Molecular Sequence Data, Mycobacterium Infections, Nontuberculous, Mycobacterium ulcerans, Phylogeny, Polymorphism, Genetic, Tandem Repeat Sequences}, issn = {1080-6040}, doi = {10.3201/eid1205.051191}, author = {Stragier, Pieter and Ablordey, Anthony and Bayonne, L Manou and Lugor, Yatta L and Sindani, Ireneaus S and Suykerbuyk, Patrick and Wabinga, Henry and Meyers, Wayne M and Portaels, Fran{\c c}oise} } @article {6, title = {Evidence for an intramacrophage growth phase of Mycobacterium ulcerans.}, journal = {Infect Immun}, volume = {75}, year = {2007}, month = {2007 Feb}, pages = {977-87}, abstract = {

Mycobacterium ulcerans is the etiologic agent of Buruli ulcer (BU), an emerging tropical skin disease. Virulent M. ulcerans secretes mycolactone, a cytotoxic exotoxin with a key pathogenic role. M. ulcerans in biopsy specimens has been described as an extracellular bacillus. In vitro assays have suggested a mycolactone-induced inhibition of M. ulcerans uptake by macrophages in which its proliferation has not been demonstrated. Therefore, and uniquely for a mycobacterium, M. ulcerans has been classified as an extracellular pathogen. In specimens from patients and in mouse footpad lesions, extracellular bacilli were concentrated in central necrotic acellular areas; however, we found bacilli within macrophages in surrounding inflammatory infiltrates. We demonstrated that mycolactone-producing M. ulcerans isolates are efficiently phagocytosed by murine macrophages, indicating that the extracellular location of M. ulcerans is not a result of inhibition of phagocytosis. Additionally, we found that M. ulcerans multiplies inside cultured mouse macrophages when low multiplicities of infection are used to prevent early mycolactone-associated cytotoxicity. Following the proliferation phase within macrophages, M. ulcerans induces the lysis of the infected host cells, becoming extracellular. Our data show that M. ulcerans, like M. tuberculosis, is an intracellular parasite with phases of intramacrophage and extracellular multiplication. The occurrence of an intramacrophage phase is in accordance with the development of cell-mediated and delayed-type hypersensitivity responses in BU patients.

}, keywords = {Animals, Bacterial Toxins, Cells, Cultured, Disease Models, Animal, Female, Foot, Histocytochemistry, Humans, Macrolides, Macrophages, Mice, Mice, Inbred BALB C, Microscopy, Electron, Transmission, Mycobacterium Infections, Nontuberculous, Mycobacterium ulcerans, Phagocytosis, Skin Diseases, Bacterial, Skin Ulcer}, issn = {0019-9567}, doi = {10.1128/IAI.00889-06}, author = {Torrado, Eg{\'\i}dio and Fraga, Alexandra G and Castro, Ant{\'o}nio G and Stragier, Pieter and Meyers, Wayne M and Portaels, Fran{\c c}oise and Silva, Manuel T and Pedrosa, Jorge} } @article {8, title = {Identification of a new variable number tandem repeat locus in Mycobacterium ulcerans for potential strain discrimination among African isolates.}, journal = {Clin Microbiol Infect}, volume = {13}, year = {2007}, month = {2007 Jul}, pages = {734-6}, abstract = {

Intra-species discrimination in the highly clonal pathogen Mycobacterium ulcerans was studied in a diverse collection of isolates by PCR amplification of a short sequence repeat locus containing heterogeneous arrays of tri-nucleotide repeats with an ACC consensus pattern. Post-amplification analysis indicated excellent typeability and identified five M. ulcerans alleles, including a unique Angolan type differentiated for the first time among a genetically conserved cluster of African isolates. These results are consistent with previously investigated independent markers, and provide an additional locus for variable number tandem repeat-based typing of M. ulcerans.

}, keywords = {Africa, Bacterial Typing Techniques, DNA, Bacterial, Humans, Minisatellite Repeats, Mycobacterium Infections, Nontuberculous, Mycobacterium ulcerans, Polymerase Chain Reaction, Sequence Alignment, Sequence Analysis, DNA, Species Specificity}, issn = {1198-743X}, doi = {10.1111/j.1469-0691.2007.01716.x}, author = {Ablordey, A and Fonteyne, P-A and Stragier, P and Vandamme, P and Portaels, F} } @article {11, title = {[Follow-up of the first case of Mycobacterium ulcerans infection documented by PCR, genotyping and culture in the Republic of Congo-Brazzaville].}, journal = {Med Trop (Mars)}, volume = {68}, year = {2008}, month = {2008 Apr}, pages = {137-43}, abstract = {

This article presents follow-up data from the first patient in whom Mycobacterium ulcerans infection (MUI) was documented by PCR, genotyping and culture in the Republic of Congo-Brazzaville. Findings show the importance of regular clinical and microbiological evaluation for the disseminated form of the disease. The patient was probably infected in Pointe Noire where MUI has been described but never documented. Culture of specimens collected before antibiotic treatment showed that the bacterium was sensitive to the antibiotics being administered (streptomycin and rifampin) and was identical to isolates from Atlantic-coast regions of West Africa where MUI is endemic. The patient was treated with streptomycin and rifampin for 12 weeks in association with surgery. During treatment clinical examination was performed every day and microbiological analysis every two weeks. The duration of follow-up from the end of specific antibiotic treatment was 26 months. Medical treatment failed to prevent bone involvement and fistulae that were treated by surgery. However medical treatment may have limited dissemination of the disease. Serial microbiological evaluation was useful to detect bone involvement in this patient, but persistent positive gene amplification is not a proof of active disease. This study confirms that MUI is still endemic in the region of Pointe Noire. This finding underlines the need to optimize epidemiologic surveillance, laboratory diagnostic capabilities, and therapeutic management in the Republic of Congo-Brazzaville.

}, keywords = {Adult, Anti-Bacterial Agents, Biopsy, Buruli Ulcer, Congo, DNA, Bacterial, Follow-Up Studies, Genotype, Humans, Male, Mycobacterium ulcerans, Polymerase Chain Reaction, Skin}, issn = {0025-682X}, author = {Kibadi, K and Stragier, P and Muyembe-Tamfum, J J and Pedrosa, J and Portaels, F} } @article {1, title = {Genetic diversity and population structure of Mycobacterium marinum: new insights into host and environmental specificities.}, journal = {J Clin Microbiol}, volume = {50}, year = {2012}, pages = {3627-34}, abstract = {

Mycobacterium marinum causes a systemic tuberculosis-like disease in fish and skin infections in humans that can spread to deeper structures, resulting in tenosynovitis, arthritis, and osteomyelitis. However, little information is available concerning (i) the intraspecific genetic diversity of M. marinum isolated from humans and animals; (ii) M. marinum genotype circulation in the different ecosystems, and (iii) the link between M. marinum genetic diversity and hosts (humans and fish). Here, we conducted a genetic study on 89 M. marinum isolates from humans (n = 68) and fish (n = 21) by using mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) typing. The results show that the M. marinum population is genetically structured not only according to the host but also according to the ecosystem as well as to tissue tropism in humans. This suggests the existence of different genetic pools in the function of the biological and ecological compartments. Moreover, the presence of only certain M. marinum genotypes in humans suggests a different zoonotic potential of the M. marinum genotypes. Considering that the infection is linked to aquarium activity, a significant genetic difference was also detected when the human tissue tropism of M. marinum was taken into consideration, with a higher genetic polymorphism in strains isolated from patients with cutaneous forms than from individuals with deeper-structure infection. It appears that only few genotypes can produce deeper infections in humans, suggesting that the immune system might play a filtering role.

}, keywords = {Adolescent, Adult, Aged, Animals, Biota, Child, Child, Preschool, DNA, Bacterial, Female, Fish Diseases, Fishes, Genetic Variation, Genotype, Humans, Interspersed Repetitive Sequences, Male, Middle Aged, Molecular Typing, Mycobacterium Infections, Nontuberculous, Mycobacterium marinum, Young Adult}, issn = {1098-660X}, doi = {10.1128/JCM.01274-12}, author = {Broutin, Vincent and Ba{\~n}uls, Anne-Laure and Aubry, Alexandra and Keck, Nicolas and Choisy, Marc and Bernardet, Jean-Fran{\c c}ois and Michel, Christian and Raymond, Jean-Christophe and Libert, C{\'e}dric and Barnaud, Antoine and Stragier, Pieter and Portaels, Fran{\c c}oise and Terru, Dominique and Belon, Claudine and Dereure, Olivier and Gutierrez, Cristina and Boschiroli, Maria-Laura and Van De Perre, Philippe and Cambau, Emmanuelle and Godreuil, Sylvain} }