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Mycobacteriology and Aerobic Actinomycetes

Genetic Diversity of Mycobacterium africanum Clinical Isolates Based on IS6110-Restriction Fragment Length Polymorphism Analysis, Spoligotyping, and Variable Number of Tandem DNA Repeats

Cristina Viana-Niero, Cristina Gutierrez, Christophe Sola, Ingrid Filliol, Fadila Boulahbal, Véronique Vincent, Nalin Rastogi
Cristina Viana-Niero
Centre National de Référence des Mycobactéries, Institut Pasteur, 75724-Paris Cedex 15, France, and
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Cristina Gutierrez
Centre National de Référence des Mycobactéries, Institut Pasteur, 75724-Paris Cedex 15, France, and
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Christophe Sola
Unité de la Tuberculose et des Mycobactéries, Institut Pasteur de Guadeloupe, F-97165 Pointe-à-Pitre Cedex, Guadeloupe
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Ingrid Filliol
Unité de la Tuberculose et des Mycobactéries, Institut Pasteur de Guadeloupe, F-97165 Pointe-à-Pitre Cedex, Guadeloupe
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Fadila Boulahbal
Centre National de Référence des Mycobactéries, Institut Pasteur, 75724-Paris Cedex 15, France, and
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Véronique Vincent
Centre National de Référence des Mycobactéries, Institut Pasteur, 75724-Paris Cedex 15, France, and
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Nalin Rastogi
Unité de la Tuberculose et des Mycobactéries, Institut Pasteur de Guadeloupe, F-97165 Pointe-à-Pitre Cedex, Guadeloupe
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DOI: 10.1128/JCM.39.1.57-65.2001
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    Fig. 1.

    Dendrogram showing the results obtained by spoligotyping of 81 M. africanum clinical isolates of various geographic origins (index, 1-Jaccard; algorithm, unweighted pair group using arithmetic average). Three subgroups (A1 to A3) were defined on the basis of clinical isolates showing a similarity index of ≥0.7. The asterisk followed by a number (from 1 to 30) after some isolate numbers designates the 10 isolates per subgroup (A1 to A3) that were randomly chosen for further analysis using VNTR. Isolates marked by an asterisk under “Origin” show identical spoligotypes between our isolates and those reported in previously published studies (13, 16, 17). Geographic origins, according to ISO code 3166, FXX, metropolitan France; SEN, Senegal; CIV, Ivory Coast; MRT, Mauritania; MDG, Madagascar; CMR, Cameroon; BEN, Benin; BFA, Burkina Faso; CAF, Central African Republic.

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    Fig. 2.

    Representative spoligotyping patterns of some of the clinical isolates of the three subgroups (A1 to A3) compared toM. tuberculosis and M. bovis, which mostly do not hybridize with spacers 33 to 36 and 39 to 43, respectively. Note thatM. africanum isolates show a specific pattern that is intermediate between those of M. tuberculosis and M. bovis. Subgroups A1 and A2 contain isolates with homogeneous spoligotyping patterns, whereas A3 constitutes a very heterogeneous group of isolates. Profiles obtained for type strains of M. tuberculosis and M. bovis are shown at the top.

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    Fig. 3.

    IS6110-RFLP genomic patterns of representative clinical isolates of the three subgroups (A1 to A3) as previously defined using spoligotyping. Note that the A1 subgroup contains significantly fewer copies of IS6110 than the A2 and A3 subgroups. Lanes M, reference strain Mt14323.

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    Fig. 4.

    Phylogenetic trees, constructed using the neighbor-joining method and the calculation of the 1-Jaccard index, for a collection of M. africanum isolates representative of subtypes A1, A2, and A3. (A) Spoligotyping-derived tree; (B) VNTR-derived tree; (C) combined numerical analysis based on spoligotyping and VNTR results. Each branch shows three levels of information; strain number (in boldface), showing the 10 isolates selected from each subgroup, which are numbered 1 to 10 for A1, 11 to 20 for A2, and 21 to 30 for A3; the major subgroups A1 to A3; and the geographic origin according to ISO code 3166 as described in the legend to Fig. 1.

Tables

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  • Table 1.

    Summary of VNTR resultsa

    SubgroupNo. of isolates with the following VNTR allele (ETR-A to ETR-E)
    6454474545323336453574535635357453642432714427253642232324324223242422
    A111123110000000
    A2000000010000000
    A300000003111211
    Total111231113111211
    • ↵a VNTR results (ETR-A to ETR-E, five-digit representation) for 30 clinical isolates of M. africanum (10 isolates randomly selected from subgroups A1 to A3) that were defined using spoligotyping and IS6110-RFLP analysis (Fig. 1 to 3).

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Genetic Diversity of Mycobacterium africanum Clinical Isolates Based on IS6110-Restriction Fragment Length Polymorphism Analysis, Spoligotyping, and Variable Number of Tandem DNA Repeats
Cristina Viana-Niero, Cristina Gutierrez, Christophe Sola, Ingrid Filliol, Fadila Boulahbal, Véronique Vincent, Nalin Rastogi
Journal of Clinical Microbiology Jan 2001, 39 (1) 57-65; DOI: 10.1128/JCM.39.1.57-65.2001

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Genetic Diversity of Mycobacterium africanum Clinical Isolates Based on IS6110-Restriction Fragment Length Polymorphism Analysis, Spoligotyping, and Variable Number of Tandem DNA Repeats
Cristina Viana-Niero, Cristina Gutierrez, Christophe Sola, Ingrid Filliol, Fadila Boulahbal, Véronique Vincent, Nalin Rastogi
Journal of Clinical Microbiology Jan 2001, 39 (1) 57-65; DOI: 10.1128/JCM.39.1.57-65.2001
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KEYWORDS

Bacterial Typing Techniques
Genetic Variation
mycobacterium
tuberculosis

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