6533b7dbfe1ef96bd1270657

RESEARCH PRODUCT

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

Ripp FabianKrombholz Christopher FelixLiu YongchaoWeber MathiasSchäfer AnneSchmidt BertilKöppel ReneHankeln Thomas

subject

MeatMethodology ArticleChromosome MappingHigh-Throughput Nucleotide SequencingSequence Analysis DNABiosurveillanceSpecies SpecificityIlluminaCalibrationDatabases GeneticFood QualityNext-generation sequencingAnimalsHumansMetagenomicsSpecies identificationReal-time PCR

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.

10.1186/1471-2164-15-639https://pubmed.ncbi.nlm.nih.gov/25081296