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All-Food-Seq (AFS): a quantifiable screen for species in biological samples by deep DNA sequencing

BACKGROUND: DNA-based methods like PCR efficiently identify and quantify the taxon composition of complex biological materials, but are limited to detecting species targeted by the choice of the primer assay. We show here how untargeted deep sequencing of foodstuff total genomic DNA, followed by bio...

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Autores principales: Ripp, Fabian, Krombholz, Christopher Felix, Liu, Yongchao, Weber, Mathias, Schäfer, Anne, Schmidt, Bertil, Köppel, Rene, Hankeln, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4131036/
https://www.ncbi.nlm.nih.gov/pubmed/25081296
http://dx.doi.org/10.1186/1471-2164-15-639
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author Ripp, Fabian
Krombholz, Christopher Felix
Liu, Yongchao
Weber, Mathias
Schäfer, Anne
Schmidt, Bertil
Köppel, Rene
Hankeln, Thomas
author_facet Ripp, Fabian
Krombholz, Christopher Felix
Liu, Yongchao
Weber, Mathias
Schäfer, Anne
Schmidt, Bertil
Köppel, Rene
Hankeln, Thomas
author_sort Ripp, Fabian
collection PubMed
description BACKGROUND: DNA-based methods like PCR efficiently identify and quantify the taxon composition of complex biological materials, but are limited to detecting species targeted by the choice of the primer assay. We show here how untargeted deep sequencing of foodstuff total genomic DNA, followed by bioinformatic analysis of sequence reads, facilitates highly accurate identification of species from all kingdoms of life, at the same time enabling quantitative measurement of the main ingredients and detection of unanticipated food components. RESULTS: Sequence data simulation and real-case Illumina sequencing of DNA from reference sausages composed of mammalian (pig, cow, horse, sheep) and avian (chicken, turkey) species are able to quantify material correctly at the 1% discrimination level via a read counting approach. An additional metagenomic step facilitates identification of traces from animal, plant and microbial DNA including unexpected species, which is prospectively important for the detection of allergens and pathogens. CONCLUSIONS: Our data suggest that deep sequencing of total genomic DNA from samples of heterogeneous taxon composition promises to be a valuable screening tool for reference species identification and quantification in biosurveillance applications like food testing, potentially alleviating some of the problems in taxon representation and quantification associated with targeted PCR-based approaches. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2164-15-639) contains supplementary material, which is available to authorized users.
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spelling pubmed-41310362014-08-18 All-Food-Seq (AFS): a quantifiable screen for species in biological samples by deep DNA sequencing Ripp, Fabian Krombholz, Christopher Felix Liu, Yongchao Weber, Mathias Schäfer, Anne Schmidt, Bertil Köppel, Rene Hankeln, Thomas BMC Genomics Methodology Article BACKGROUND: DNA-based methods like PCR efficiently identify and quantify the taxon composition of complex biological materials, but are limited to detecting species targeted by the choice of the primer assay. We show here how untargeted deep sequencing of foodstuff total genomic DNA, followed by bioinformatic analysis of sequence reads, facilitates highly accurate identification of species from all kingdoms of life, at the same time enabling quantitative measurement of the main ingredients and detection of unanticipated food components. RESULTS: Sequence data simulation and real-case Illumina sequencing of DNA from reference sausages composed of mammalian (pig, cow, horse, sheep) and avian (chicken, turkey) species are able to quantify material correctly at the 1% discrimination level via a read counting approach. An additional metagenomic step facilitates identification of traces from animal, plant and microbial DNA including unexpected species, which is prospectively important for the detection of allergens and pathogens. CONCLUSIONS: Our data suggest that deep sequencing of total genomic DNA from samples of heterogeneous taxon composition promises to be a valuable screening tool for reference species identification and quantification in biosurveillance applications like food testing, potentially alleviating some of the problems in taxon representation and quantification associated with targeted PCR-based approaches. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1471-2164-15-639) contains supplementary material, which is available to authorized users. BioMed Central 2014-07-31 /pmc/articles/PMC4131036/ /pubmed/25081296 http://dx.doi.org/10.1186/1471-2164-15-639 Text en © Ripp et al.; licensee BioMed Central Ltd. 2014 This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.
spellingShingle Methodology Article
Ripp, Fabian
Krombholz, Christopher Felix
Liu, Yongchao
Weber, Mathias
Schäfer, Anne
Schmidt, Bertil
Köppel, Rene
Hankeln, Thomas
All-Food-Seq (AFS): a quantifiable screen for species in biological samples by deep DNA sequencing
title All-Food-Seq (AFS): a quantifiable screen for species in biological samples by deep DNA sequencing
title_full All-Food-Seq (AFS): a quantifiable screen for species in biological samples by deep DNA sequencing
title_fullStr All-Food-Seq (AFS): a quantifiable screen for species in biological samples by deep DNA sequencing
title_full_unstemmed All-Food-Seq (AFS): a quantifiable screen for species in biological samples by deep DNA sequencing
title_short All-Food-Seq (AFS): a quantifiable screen for species in biological samples by deep DNA sequencing
title_sort all-food-seq (afs): a quantifiable screen for species in biological samples by deep dna sequencing
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4131036/
https://www.ncbi.nlm.nih.gov/pubmed/25081296
http://dx.doi.org/10.1186/1471-2164-15-639
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