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Evaluation of Direct 16S rDNA sequencing as a metagenomics-based approach to screening bacteria in bottled water
DTU Technical University of Denmark, Denmark.ORCID iD: 0000-0001-6492-1245
2013 (English)In: Biosecurity and bioterrorism, ISSN 1538-7135, E-ISSN 1557-850X, Vol. 11, no SUPPL. 1, p. S158-S165Article in journal (Refereed) Published
Abstract [en]

Deliberate or accidental contamination of food, feed, and water supplies poses a threat to human health worldwide. A rapid and sensitive detection technique that could replace the current labor-intensive and time-consuming culture-based methods is highly desirable. In addition to species-specific assays, such as PCR, there is a need for generic methods to screen for unknown pathogenic microorganisms in samples. This work presents a metagenomics-based direct-sequencing approach for detecting unknown microorganisms, using Bacillus cereus (as a model organism for B. anthracis) in bottled water as an example. Total DNA extraction and 16S rDNA gene sequencing were used in combination with principle component analysis and multicurve resolution to study detection level and possibility for identification. Results showed a detection level of 10 5 to 106 CFU/L. Using this method, it was possible to separate 2 B. cereus strains by the principal component plot, despite the close sequence resemblance. A linear correlation between the artificial contamination level and the relative amount of the Bacillus artificial contaminant in the metagenome was observed, and a relative amount value above 0.5 confirmed the presence of Bacillus. The analysis also revealed that background flora in the bottled water varied between the different water types that were included in the study. This method has the potential to be adapted to other biological matrices and bacterial pathogens for fast screening of unknown bacterial threats in outbreak situations. © 2013, Mary Ann Liebert, Inc.

Place, publisher, year, edition, pages
2013. Vol. 11, no SUPPL. 1, p. S158-S165
Keywords [en]
drinking water, ribosome DNA, article, Bacillus cereus, biological warfare, DNA sequence, gene amplification, genetics, human, isolation and purification, metagenomics, methodology, microbiology, polymerase chain reaction, principal component analysis, Bioterrorism, DNA, Ribosomal, Humans, Sequence Analysis, DNA
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Natural Sciences
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URN: urn:nbn:se:ri:diva-39009DOI: 10.1089/bsp.2012.0073Scopus ID: 2-s2.0-84883255803OAI: oai:DiVA.org:ri-39009DiVA, id: diva2:1324955
Available from: 2019-06-14 Created: 2019-06-14 Last updated: 2019-06-14Bibliographically approved

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Löfström, Charlotta

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