| Title: | Fast and accurate multiplex identification and quantification of seven genetically modified soybean lines using six-color digital PCR |
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| Authors: | ID Bogožalec Košir, Alexandra (Author)
ID Muller, Sabine (Author)
ID Žel, Jana (Author)
ID Milavec, Mojca (Author)
ID Mallory, Allison C. (Author)
ID Dobnik, David (Author) |
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| Files: |  URL - Source URL, visit https://doi.org/10.3390/foods12224156
 PDF - Presentation file, download (1,83 MB)
MD5: 330DC3B84B961FAC517B5F92875FD051
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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: | The proliferation of genetically modified organisms (GMOs) presents challenges to GMO testing laboratories and policymakers. Traditional methods, like quantitative real-time PCR (qPCR), face limitations in quantifying the increasing number of GMOs in a single sample. Digital PCR (dPCR), specifically multiplexing, offers a solution by enabling simultaneous quantification of multiple GMO targets. This study explores the use of the Naica six-color Crystal dPCR platform for quantifying five GM soybean lines within a single six-plex assay. Two four-color assays were also developed for added flexibility. These assays demonstrated high specificity, sensitivity (limit of detection or LOD < 25 copies per reaction) and precision (bias to an estimated copy number concentration <15%). Additionally, two approaches for the optimization of data analysis were implemented. By applying a limit-of-blank (LOB) correction, the limit of quantification (LOQ) and LOD could be more precisely determined. Pooling of reactions additionally lowered the LOD, with a two- to eight-fold increase in sensitivity. Real-life samples from routine testing were used to confirm the assays’ applicability for quantifying GM soybean lines in complex samples. This study showcases the potential of the six-color Crystal dPCR platform to revolutionize GMO testing, facilitating comprehensive analysis of GMOs in complex samples. |
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| Keywords: | digital PCR, dPCR, quantification, multiplexing, genetically modified organisms, 6-color system, virus diagnostics, virology |
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| Publication status: | Published |
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| Publication version: | Version of Record |
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| Publication date: | 01.11.2023 |
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| Year of publishing: | 2023 |
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| Number of pages: | str. [1]-17 |
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| Numbering: | iss. 22, art. no. ǂ4156 |
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| PID: | 20.500.12556/DiRROS-18583  |
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| UDC: | 578 |
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| ISSN on article: | 2304-8158 |
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| DOI: | 10.3390/foods12224156 |
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| COBISS.SI-ID: | 173360131 |
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| Note: | Soavtorji: Sabine Muller, Jana Žel, Mojca Milavec, Allison C. Mallory and David Dobnik;
Nasl. z nasl. zaslona;
Opis vira z dne 23. 11. 2023;
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| Publication date in DiRROS: | 29.03.2024 |
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| Views: | 598 |
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| Downloads: | 230 |
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