ArticleApplied and environmental microbiology2010
High-resolution in situ genotyping of Legionella pneumophila populations in drinking water by multiple-locus variable-number tandem-repeat analysis using environmental DNA.
Article in Applied and environmental microbiology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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14 citing papers in PubMed, 31 citations in OpenAlex.
- Quantitative microbial risk assessment ofNew microbes and new infections · 2025Article
- On-site filtration of large sample volumes improves the detection of opportunistic pathogens in drinking water distribution systems.Applied and environmental microbiology · 2024Article
- Chasing Waterborne Pathogens in Antarctic Human-Made and Natural Environments, with Special Reference toApplied and environmental microbiology · 2021Article
- Asymptomatic Carriage ofFrontiers in microbiology · 2021Article
- Biogeography and Environmental Drivers ofPathogens (Basel, Switzerland) · 2020Article
- Antimicrobial agent susceptibilities of Legionella pneumophila MLVA-8 genotypes.Scientific reports · 2019Article
- Methodological approaches for monitoring opportunistic pathogens in premise plumbing: A review.Water research · 2017Review
- Temperature-Dependent Growth Modeling of Environmental and Clinical Legionella pneumophila Multilocus Variable-Number Tandem-Repeat Analysis (MLVA) Genotypes.Applied and environmental microbiology · 2017Article
- Development of a genus-specific next generation sequencing approach for sensitive and quantitative determination of the Legionella microbiome in freshwater systems.BMC microbiology · 2017Article
- Comparison of sputum microbiome of legionellosis-associated patients and other pneumonia patients: indications for polybacterial infections.Scientific reports · 2017Article
- Legionella species diversity and dynamics from surface reservoir to tap water: from cold adaptation to thermophily.The ISME journal · 2016Article
- Dendrimers and Polyamino-Phenolic Ligands: Activity of New Molecules Against Legionella pneumophila Biofilms.Frontiers in microbiology · 2016Article
- Multilocus variable-number tandem-repeat analysis of Neisseria meningitidis serogroup C in China.Epidemiology and infection · 2015Article
- Assessing the viability of bacterial species in drinking water by combined cellular and molecular analyses.Microbial ecology · 2012Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Central to the understanding of infections by the waterborne pathogen Legionella pneumophila is its detection at the clonal level. Currently, multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) of L. pneumophila isolates can be used as a tool for high-resolution genotyping. Since L. pneumophila is difficult to isolate, the isolation of outbreak strains often fails due to a viable but nonculturable (VBNC) state of the respective environmental population. Therefore, we developed a cultivation-independent approach to detect single clones in drinking water. This approach is based on the extraction of DNA from drinking water followed by PCR using a set of eight VNTR primer pairs necessary for MLVA genotyping of L. pneumophila. The PCR amplicons were analyzed by single-strand conformation polymorphism (SSCP) and capillary electrophoresis to obtain the respective MLVA profiles. Parallel to the high-resolution analysis, we used the same environmental DNA to quantify the number of L. pneumophila cells in drinking water using real-time PCR with 16S rRNA gene-targeted primers. We used a set of drinking water samples from a small-scale drinking water network to test our approach. With these samples we demonstrated that the developed approach was directly applicable to DNA obtained from drinking water. We were able to detect more L. pneumophila MLVA genotypes in drinking water than we could detect by isolation. Our approach could be a valuable tool to identify outbreak strains even after the outbreak has occurred and has the potential to be applied directly to clinical material.
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