Targeting the rpoB gene using nested PCR-restriction fragment length polymorphism for identification of nontuberculous mycobacteria in hospital tap water

Shin, Ji‐Hyun; Lee, Haekyung; Cho, Eunjin; Yu, Jae-Yon; Kang, Yeon‐Ho

doi:10.1007/s12275-008-0102-6

Cited by 18 publications

(16 citation statements)

References 40 publications

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“…Similarly, a high diversity of mycobacteria, some of which, such as M. avium, M. intracellulare, M. kansasii, M. abscessus, and M. chelonae (33,47,65,73,77), are known to be opportunistic pathogens, has been demonstrated to be characteristic of the drinking water environment; among those species, M. avium is currently on the U.S. EPA candidate contaminant list. In the present study, a considerable number of mycobacteria were detected, accounting for 0.1% to 68% of total bacterial estimate, assuming that each bacterium has a single copy of the 16S rRNA gene.…”

Section: Discussionmentioning

confidence: 99%

Molecular Survey of the Occurrence of Legionella spp., Mycobacterium spp., Pseudomonas aeruginosa, and Amoeba Hosts in Two Chloraminated Drinking Water Distribution Systems

Wang

Edwards

Falkinham

et al. 2012

Appl Environ Microbiol

251

230

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ABSTRACTThe spread of opportunistic pathogens via public water systems is of growing concern. The purpose of this study was to identify patterns of occurrence among three opportunistic pathogens (Legionella pneumophila,Mycobacterium avium, andPseudomonas aeruginosa) relative to biotic and abiotic factors in two representative chloraminated drinking water distribution systems using culture-independent methods. Generally, a high occurrence ofLegionella(≥69.0%) and mycobacteria (100%), lower occurrence ofL. pneumophila(≤20%) andM. avium(≤33.3%), and rare detection ofPseudomonas aeruginosa(≤13.3%) were observed in both systems according to quantitative PCR. Also,Hartmanella vermiformiswas more prevalent thanAcanthamoeba, both of which are known hosts for opportunistic pathogen amplification, the latter itself containing pathogenic members. Three-minute flushing served to distinguish distribution system water from plumbing in buildings (i.e., premise plumbing water) and resulted in reduced numbers of copies ofLegionella, mycobacteria,H. vermiformis, and 16S rRNA genes (P< 0.05) while yielding distinct terminal restriction fragment polymorphism (T-RFLP) profiles of 16S rRNA genes. Within certain subgroups of samples, some positive correlations, including correlations of numbers of mycobacteria and total bacteria (16S rRNA genes),H. vermiformisand total bacteria, mycobacteria andH. vermiformis, andLegionellaandH. vermiformis, were noted, emphasizing potential microbial ecological relationships. Overall, the results provide insight into factors that may aid in controlling opportunistic pathogen proliferation in real-world water systems.

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Section: Discussionmentioning

confidence: 99%

Molecular Survey of the Occurrence of Legionella spp., Mycobacterium spp., Pseudomonas aeruginosa, and Amoeba Hosts in Two Chloraminated Drinking Water Distribution Systems

Wang

Edwards

Falkinham

et al. 2012

Appl Environ Microbiol

251

230

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“…The other objective of our study was to evaluate the use of rpoB gene sequencing for the molecular identification of isolates and discrimination between the different groups of M. abscessus sensu lato. Several recent studies have questioned the reliability of approaches based solely on rpoB sequencing (31,34,47,48). This is because of the possible horizontal transfer of the rpoB gene between the different groups of M. abscessus sensu lato, especially from the M. abscessus group to the M. massiliense group.…”

Section: Discussionmentioning

confidence: 99%

“…massiliense and M. bolletii were characterized as new species distinct from M. abscessus on the basis of their rpoB sequences (Ͼ3% sequence divergence) (2,5). Partial rpoB sequencing is now the gold standard for the molecular identification of the three species (3,4,16,38,48). However, recent studies have highlighted the inaccuracy of single-target sequencing, including rpoB sequencing, for distinguishing between M. abscessus, M. massiliense, and M. bolletii (18,31,36,59).…”

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confidence: 99%

Multilocus Sequence Analysis and rpoB Sequencing of Mycobacterium abscessus (Sensu Lato) Strains

et al. 2011

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Mycobacterium abscessus, Mycobacterium bolletii, and Mycobacterium massiliense (Mycobacterium abscessus sensu lato) are closely related species that currently are identified by the sequencing of the rpoB gene. However, recent studies show that rpoB sequencing alone is insufficient to discriminate between these species, and some authors have questioned their current taxonomic classification. We studied here a large collection of M. abscessus (sensu lato) strains by partial rpoB sequencing (752 bp) and multilocus sequence analysis (MLSA). The final MLSA scheme developed was based on the partial sequences of eight housekeeping genes: argH, cya, glpK, gnd, murC, pgm, pta, and purH. The strains studied included the three type strains (M. abscessus CIP 104536 T , M. massiliense CIP 108297 T , and M. bolletii CIP 108541 T ) and 120 isolates recovered between 1997 and 2007 in France, Germany, Switzerland, and Brazil. The rpoB phylogenetic tree confirmed the existence of three main clusters, each comprising the type strain of one species. However, divergence values between the M. massiliense and M. bolletii clusters all were below 3% and between the M. abscessus and M. massiliense clusters were from 2.66 to 3.59%. The tree produced using the concatenated MLSA gene sequences (4,071 bp) also showed three main clusters, each comprising the type strain of one species. The M. abscessus cluster had a bootstrap value of 100% and was mostly compact. Bootstrap values for the M. massiliense and M. bolletii branches were much lower (71 and 61%, respectively), with the M. massiliense cluster having a fuzzy aspect. Mean (range) divergence values were 2.17% (1.13 to 2.58%) between the M. abscessus and M. massiliense clusters, 2.37% (1.5 to 2.85%) between the M. abscessus and M. bolletii clusters, and 2.28% (0.86 to 2.68%) between the M. massiliense and M. bolletii clusters. Adding the rpoB sequence to the MLSA-concatenated sequence (total sequence, 4,823 bp) had little effect on the clustering of strains. We found 10/120 (8.3%) isolates for which the concatenated MLSA gene sequence and rpoB sequence were discordant (e.g., M. massiliense MLSA sequence and M. abscessus rpoB sequence), suggesting the intergroup lateral transfers of rpoB. In conclusion, our study strongly supports the recent proposal that M. abscessus, M. massiliense, and M. bolletii should constitute a single species. Our findings also indicate that there has been a horizontal transfer of rpoB sequences between these subgroups, precluding the use of rpoB sequencing alone for the accurate identification of the two proposed M. abscessus subspecies.

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“…RGM are commonly recovered from water treatment and distribution systems (31)(32)(33)(34)(35)(36)(37)(38), probably because they can form biofilms (2,(39)(40)(41) and resist chlorination and oligotrophic conditions (42,43). Mycobacterium chelonae and Mycobacterium fortuitum were the species most frequently detected in previous studies (31,34,35,40,44).…”

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confidence: 95%

Diversity, Community Composition, and Dynamics of Nonpigmented and Late-Pigmenting Rapidly Growing Mycobacteria in an Urban Tap Water Production and Distribution System

Dubrou¹,

Konjek

Macheras

et al. 2013

Appl Environ Microbiol

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e Nonpigmented and late-pigmenting rapidly growing mycobacteria (RGM) have been reported to commonly colonize water production and distribution systems. However, there is little information about the nature and distribution of RGM species within the different parts of such complex networks or about their clustering into specific RGM species communities. We conducted a large-scale survey between 2007 and 2009 in the Parisian urban tap water production and distribution system. We analyzed 1,418 water samples from 36 sites, covering all production units, water storage tanks, and distribution units; RGM isolates were identified by using rpoB gene sequencing. We detected 18 RGM species and putative new species, with most isolates being Mycobacterium chelonae and Mycobacterium llatzerense. Using hierarchical clustering and principal-component analysis, we found that RGM were organized into various communities correlating with water origin (groundwater or surface water) and location within the distribution network. Water treatment plants were more specifically associated with species of the Mycobacterium septicum group. On average, M. chelonae dominated network sites fed by surface water, and M. llatzerense dominated those fed by groundwater. Overall, the M. chelonae prevalence index increased along the distribution network and was associated with a correlative decrease in the prevalence index of M. llatzerense, suggesting competitive or niche exclusion between these two dominant species. Our data describe the great diversity and complexity of RGM species living in the interconnected environments that constitute the water production and distribution system of a large city and highlight the prevalence index of the potentially pathogenic species M. chelonae in the distribution network. N onpigmented and late-pigmenting rapidly growing mycobacteria (RGM) are ubiquitous in soil and water environments (1-6). Most are harmless saprophytes, but some, such as Mycobacterium chelonae and Mycobacterium abscessus, are opportunistic pathogens that are causing increasing concern (7-9). Potentially pathogenic RGM are associated with a wide spectrum of diseases in humans, including pulmonary tract, skin, soft tissue, and disseminated infections (10-14), mostly in patients with predisposing conditions (e.g., lung diseases, chronic obstructive pulmonary disease, cystic fibrosis, genetic predisposition, or immunosuppressive therapy). Contamination and hypersensitive reactions are often due to environmental exposure (associated with, for example, hot tubs, metalworking fluids, or contaminated dust) (15-19), and many outbreaks following invasive medical procedures have been reported over the last decade (20-25). Tap water may be an important source of contamination in urban environments (13,18,(26)(27)(28)(29)(30).RGM are commonly recovered from water treatment and distribution systems (31-38), probably because they can form biofilms (2, 39-41) and resist chlorination and oligotrophic conditions (42,43). Mycobacterium chelonae and Mycobact...

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Targeting the rpoB gene using nested PCR-restriction fragment length polymorphism for identification of nontuberculous mycobacteria in hospital tap water

Cited by 18 publications

References 40 publications

Molecular Survey of the Occurrence of Legionella spp., Mycobacterium spp., Pseudomonas aeruginosa, and Amoeba Hosts in Two Chloraminated Drinking Water Distribution Systems

Molecular Survey of the Occurrence of Legionella spp., Mycobacterium spp., Pseudomonas aeruginosa, and Amoeba Hosts in Two Chloraminated Drinking Water Distribution Systems

Multilocus Sequence Analysis and rpoB Sequencing of Mycobacterium abscessus (Sensu Lato) Strains

Diversity, Community Composition, and Dynamics of Nonpigmented and Late-Pigmenting Rapidly Growing Mycobacteria in an Urban Tap Water Production and Distribution System

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