Interlaboratory Validation of a DNA Metabarcoding Assay for Mammalian and Poultry Species to Detect Food Adulteration
Meat species authentication in food is most commonly based on the detection of genetic variations. Official food control laboratories frequently apply single and multiplex real-time polymerase chain reaction (PCR) assays and/or DNA arrays. However, in the near future, DNA metabarcoding, the generati...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-04-01
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| Series: | Foods |
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| Online Access: | https://www.mdpi.com/2304-8158/11/8/1108 |
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| author | Stefanie Dobrovolny Steffen Uhlig Kirstin Frost Anja Schlierf Kapil Nichani Kirsten Simon Margit Cichna-Markl Rupert Hochegger |
| author_facet | Stefanie Dobrovolny Steffen Uhlig Kirstin Frost Anja Schlierf Kapil Nichani Kirsten Simon Margit Cichna-Markl Rupert Hochegger |
| author_sort | Stefanie Dobrovolny |
| collection | DOAJ |
| description | Meat species authentication in food is most commonly based on the detection of genetic variations. Official food control laboratories frequently apply single and multiplex real-time polymerase chain reaction (PCR) assays and/or DNA arrays. However, in the near future, DNA metabarcoding, the generation of PCR products for DNA barcodes, followed by massively parallel sequencing by next generation sequencing (NGS) technologies, could be an attractive alternative. DNA metabarcoding is superior to well-established methodologies since it allows simultaneous identification of a wide variety of species not only in individual foodstuffs but even in complex mixtures. We have recently published a DNA metabarcoding assay for the identification and differentiation of 15 mammalian species and six poultry species. With the aim to harmonize analytical methods for food authentication across EU Member States, the DNA metabarcoding assay has been tested in an interlaboratory ring trial including 15 laboratories. Each laboratory analyzed 16 anonymously labelled samples (eight samples, two subsamples each), comprising six DNA extract mixtures, one DNA extract from a model sausage, and one DNA extract from maize (negative control). Evaluation of data on repeatability, reproducibility, robustness, and measurement uncertainty indicated that the DNA metabarcoding method is applicable for meat species authentication in routine analysis. |
| first_indexed | 2024-03-09T10:36:22Z |
| format | Article |
| id | doaj.art-91f30c0cd898429a8a7eecff0231bd4f |
| institution | Directory Open Access Journal |
| issn | 2304-8158 |
| language | English |
| last_indexed | 2024-03-09T10:36:22Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Foods |
| spelling | doaj.art-91f30c0cd898429a8a7eecff0231bd4f2023-12-01T20:53:43ZengMDPI AGFoods2304-81582022-04-01118110810.3390/foods11081108Interlaboratory Validation of a DNA Metabarcoding Assay for Mammalian and Poultry Species to Detect Food AdulterationStefanie Dobrovolny0Steffen Uhlig1Kirstin Frost2Anja Schlierf3Kapil Nichani4Kirsten Simon5Margit Cichna-Markl6Rupert Hochegger7Austrian Agency for Health and Food Safety (AGES), Department for Molecular Biology and Microbiology, Institute for Food Safety Vienna, Spargelfeldstrasse 191, 1220 Vienna, AustriaQuoData GmbH, Prellerstrasse 14, 01309 Dresden, GermanyQuoData GmbH, Prellerstrasse 14, 01309 Dresden, GermanyQuoData GmbH, Prellerstrasse 14, 01309 Dresden, GermanyQuoData GmbH, Prellerstrasse 14, 01309 Dresden, GermanyQuoData GmbH, Prellerstrasse 14, 01309 Dresden, GermanyDepartment of Analytical Chemistry, Faculty of Chemistry, University of Vienna, Währinger Strasse 38, 1090 Vienna, AustriaAustrian Agency for Health and Food Safety (AGES), Department for Molecular Biology and Microbiology, Institute for Food Safety Vienna, Spargelfeldstrasse 191, 1220 Vienna, AustriaMeat species authentication in food is most commonly based on the detection of genetic variations. Official food control laboratories frequently apply single and multiplex real-time polymerase chain reaction (PCR) assays and/or DNA arrays. However, in the near future, DNA metabarcoding, the generation of PCR products for DNA barcodes, followed by massively parallel sequencing by next generation sequencing (NGS) technologies, could be an attractive alternative. DNA metabarcoding is superior to well-established methodologies since it allows simultaneous identification of a wide variety of species not only in individual foodstuffs but even in complex mixtures. We have recently published a DNA metabarcoding assay for the identification and differentiation of 15 mammalian species and six poultry species. With the aim to harmonize analytical methods for food authentication across EU Member States, the DNA metabarcoding assay has been tested in an interlaboratory ring trial including 15 laboratories. Each laboratory analyzed 16 anonymously labelled samples (eight samples, two subsamples each), comprising six DNA extract mixtures, one DNA extract from a model sausage, and one DNA extract from maize (negative control). Evaluation of data on repeatability, reproducibility, robustness, and measurement uncertainty indicated that the DNA metabarcoding method is applicable for meat species authentication in routine analysis.https://www.mdpi.com/2304-8158/11/8/1108DNA metabarcodinganimal speciesspecies identificationNGSfood adulterationvalidation |
| spellingShingle | Stefanie Dobrovolny Steffen Uhlig Kirstin Frost Anja Schlierf Kapil Nichani Kirsten Simon Margit Cichna-Markl Rupert Hochegger Interlaboratory Validation of a DNA Metabarcoding Assay for Mammalian and Poultry Species to Detect Food Adulteration Foods DNA metabarcoding animal species species identification NGS food adulteration validation |
| title | Interlaboratory Validation of a DNA Metabarcoding Assay for Mammalian and Poultry Species to Detect Food Adulteration |
| title_full | Interlaboratory Validation of a DNA Metabarcoding Assay for Mammalian and Poultry Species to Detect Food Adulteration |
| title_fullStr | Interlaboratory Validation of a DNA Metabarcoding Assay for Mammalian and Poultry Species to Detect Food Adulteration |
| title_full_unstemmed | Interlaboratory Validation of a DNA Metabarcoding Assay for Mammalian and Poultry Species to Detect Food Adulteration |
| title_short | Interlaboratory Validation of a DNA Metabarcoding Assay for Mammalian and Poultry Species to Detect Food Adulteration |
| title_sort | interlaboratory validation of a dna metabarcoding assay for mammalian and poultry species to detect food adulteration |
| topic | DNA metabarcoding animal species species identification NGS food adulteration validation |
| url | https://www.mdpi.com/2304-8158/11/8/1108 |
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