| Title: | Quantitative analysis of food and feed samples with droplet digital PCR |
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| Authors: | ID Morisset, Dany (Author)
ID Štebih, Dejan (Author)
ID Milavec, Mojca (Author)
ID Gruden, Kristina (Author)
ID Žel, Jana (Author) |
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| Files: |  URL - Source URL, visit http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0062583;jsessionid=DF4F7162FA7C190252A6F73DBD180ECA
 PDF - Presentation file, download (357,00 KB)
MD5: AA586C8A0317BEE867A5B42590EEC1C9
URL - Source URL, visit https://doi.org/10.1371/journal.pone.0062583
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| Language: | English |
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| Typology: | 1.01 - Original Scientific Article |
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| Organization: |  NIB - National Institute of Biology
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| Abstract: | In this study, the applicability of droplet digital PCR (ddPCR) for routine analysis in food and feed samples was demonstrated with the quantification of genetically modified organisms (GMOs). Real-time quantitative polymerase chain reaction (qPCR) is currently used for quantitative molecular analysis of the presence of GMOs in products. However, its use is limited for detecting and quantifying very small numbers of DNA targets, as in some complex food and feed matrices. Using ddPCR duplex assay, we have measured the absolute numbers of MON810 transgene and hmg maize reference gene copies in DNA samples. Key performance parameters of the assay were determined. The ddPCR system is shown to offer precise absolute and relative quantification of targets, without the need for calibration curves. The sensitivity (five target DNA copies) of the ddPCR assay compares well with those of individual qPCR assays and of the chamber digital PCR (cdPCR) approach. It offers a dynamic range over four orders of magnitude, greater than that of cdPCR. Moreover, when compared to qPCR, the ddPCR assay showed better repeatability at low target concentrations and a greater tolerance to inhibitors. Finally, ddPCR throughput and cost are advantageous relative to those of qPCR for routine GMO quantification. It is thus concluded that ddPCR technology can be applied for routine quantification of GMOs, or any other domain where quantitative analysis of food and feed samples is needed.
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| Keywords: | droplet digital PCR, ddPCR, genetically modified organisms |
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| Publication status: | Published |
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| Publication version: | Version of Record |
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| Publication date: | 02.05.2013 |
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| Year of publishing: | 2013 |
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| Number of pages: | str. e62583-1-e62583-9 |
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| Numbering: | Vol. 8, issue 5 |
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| PID: | 20.500.12556/DiRROS-21609  |
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| UDC: | 577.2 |
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| ISSN on article: | 1932-6203 |
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| DOI: | 10.1371/journal.pone.0062583 |
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| COBISS.SI-ID: | 2785359 |
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| Note: | Soavtorji: Dejan Štebih, Mojca Milavec, Kristina Gruden, Jana Žel;
Popis vira z dne 13. 5. 2013;
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| Publication date in DiRROS: | 04.03.2025 |
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| Views: | 545 |
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| Downloads: | 508 |
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