Evaluation of the relatedness of strains ofMycobacterium avium using pulsed-field gel electrophoresis

Burki, D; Bernasconi, C.; Bodmer, Thomas; Telenti, Amalio

doi:10.1007/bf02310358

Cited by 31 publications

(23 citation statements)

References 26 publications

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“…PFGE and AFLP analysis of M. kansasii are particularly useful because they reveal polymorphisms within each cluster. Mycobacterial strain characterization by PFGE has been previously demonstrated to be useful for epidemiological studies to trace sources of contamination and delineate outbreaks (4,8,16,29,34). Variations in the array of LRFs in both size and number may be due to sequence rearrangements, insertion or deletion of DNA, or base substitution within the restriction sites (25).…”

Section: Discussionmentioning

confidence: 99%

Genotypic characterization of five subspecies of Mycobacterium kansasii

et al. 1997

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Different molecular typing methods including restriction fragment length polymorphism (RFLP) analysis with the major polymorphic tandem repeat (MPTR) probe and the IS1652 probe, pulsed-field gel electrophoresis (PFGE), amplified fragment length polymorphism (AFLP) analysis, and PCR restriction analysis of the hsp-65 gene (PRA) were applied to clinical and water isolates of Mycobacterium kansasii. RFLP with the MPTR probe, PRA, PFGE, and AFLP analysis revealed five homogeneous clusters which appeared to be subspecies. RFLP with the MPTR probe and PRA gave patterns specific for each cluster, whereas PFGE and AFLP analysis gave polymorphic patterns. IS1652 was present in two of the five clusters and provided polymorphic patterns for one cluster only. The two IS1652-positive clusters were Accuprobe negative (Accuprobe test; Gen-Probe Inc.), and only two other clusters were Accuprobe positive. A PCR test based on the detection of a species-specific fragment (M. Yang, B. C. Ross, and B. Dwyer, J. Clin. Microbiol. 31:2769-2772, 1993) was positive for all M. kansasii strains. This PCR test is an accurate, rapid, and specific M. kansasii identification test. No subspecies was particularly more virulent, because all clusters contained clinical strains, from AIDS patients and non-AIDS patients, and environmental strains. Mycobacterium kansasii, like M. avium and M. xenopi, is one of the major pulmonary mycobacterial pathogens. The relative incidence of disease due to these pathogens varies between geographical regions. In the United States, M. avium and M. kansasii are the most frequently isolated mycobacteria, whereas M. kansasii is rare in Australia. However, in some European countries, M. kansasii and M. xenopi infections were more common than M. avium infections before the AIDS epidemic (7). M. kansasii infections are often clustered, mainly in urban areas, as reported in studies in Texas, the Czech Republic, and other regions (7, 11). M. avium predominates in human immunodeficiency virus (HIV)-infected patients, and M. avium infection represents the most frequent disseminated infection diagnosed in these patients. However, HIV-infected patients also suffer pulmonary or disseminated infections due to M. kansasii. In some areas, for example, Switzerland, the prevalence of mycobacterial disease due to M. kansasii has been unchanged by the AIDS epidemic, which was not associated with a shift to a predominance of M. avium (7). Unlike tubercle bacilli, human-to-human transmission has never been established for infections due to other mycobacteria. These mycobacterial infections are considered to be acquired from the environment. M. kansasii has been almost exclusively recovered from tap water; other environmental samples are rarely positive (3, 6, 10, 14, 23, 31). M. kansasii was found to be able to survive in tap water for up to 12 months, whereas long-term survival in soil could not be shown (10). Because the M. kansasii isolates found in drinking-water distribution systems are not contaminants from another source but are ...

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Section: Discussionmentioning

confidence: 99%

Genotypic characterization of five subspecies of Mycobacterium kansasii

et al. 1997

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“…Mycobacterial DNA plugs were prepared as previously described (54) with the following modifications. Ampicillin and D-cycloserine were added to the culture 24 h prior to harvesting at final concentrations of 0.1 and 1.0 mg/ml, respectively (8). The step requiring vortexing of the cells in the presence of 3-mm glass beads was omitted, and the Bio-Rad Genepath wash solution was replaced with TE buffer (10 mM Tris, 1 mM EDTA [pH 8.0]).…”

Section: Methodsmentioning

confidence: 99%

Comparative Genetic Analysis of Mycobacterium ulcerans and Mycobacterium marinum Reveals Evidence of Recent Divergence

et al. 2000

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Previous studies of the 16S rRNA genes from Mycobacterium ulcerans and Mycobacterium marinum have suggested a very close genetic relationship between these species (99.6% identity). However, these organisms are phenotypically distinct and cause diseases with very different pathologies. To investigate this apparent paradox, we compared 3,306 nucleotides from the partial sequences of eight housekeeping and structural genes derived from 18 M. ulcerans strains and 22 M. marinum strains. This analysis confirmed the close genetic relationship inferred from the 16S rRNA data, with nucleotide sequence identity ranging from 98.1 to 99.7%. The multilocus sequence analysis also confirmed previous genotype studies of M. ulcerans that have identified distinct genotypes within a geographical region. Single isolates of both M. ulcerans and M. marinum that were shown by the sequence analysis to be the most closely related were then selected for further study. One-and two-dimensional pulsed-field gel electrophoresis was employed to compare the architecture and size of the genome from each species. Genome sizes of approximately 4.4 and 4.6 Mb were obtained for M. ulcerans and M. marinum, respectively. Significant macrorestriction fragment polymorphism was observed between the species. However, hybridization analysis of DNA cleaved with more frequently cutting enzymes identified significant preservation of the flanking sequence at seven of the eight loci sequenced. The exception was the 16S rRNA locus. Two high-copy-number insertion sequences, IS2404 and IS2606, have recently been reported in M. ulcerans, and significantly, these elements are not present in M. marinum. Hybridization of the AseI restriction fragments from M. ulcerans with IS2404 and IS2606 indicated widespread genome distribution for both of these repeated sequences. Taken together, these data strongly suggest that M. ulcerans has recently diverged from M. marinum by the acquisition and concomitant loss of DNA in a manner analogous to the emergence of M. tuberculosis, where species diversity is being driven mainly by the activity of mobile DNA elements.

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“…Using a DNA probe consisting of IS900 and linked chromosomal DNA, it was shown that European and U.S. M. avium isolates share an RFLP pattern whereas isolates from Africa share a different pattern (398). LRF patterns have also been used as a quality control tool in a clinical mycobacteriology laboratory and as a tool for distinguishing true outbreaks of M. avium complex infection from pseudo-outbreaks due to laboratory-associated contamination (63).…”

Section: Characteristics Of the M Avium Complexmentioning

confidence: 99%