| Naslov: | Fast and accurate multiplex identification and quantification of seven genetically modified soybean lines using six-color digital PCR |
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| Avtorji: | ID Bogožalec Košir, Alexandra (Avtor)
ID Muller, Sabine (Avtor)
ID Žel, Jana (Avtor)
ID Milavec, Mojca (Avtor)
ID Mallory, Allison C. (Avtor)
ID Dobnik, David (Avtor) |
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| Datoteke: |  URL - Izvorni URL, za dostop obiščite https://doi.org/10.3390/foods12224156
 PDF - Izvorna datoteka, prenos (1,83 MB)
MD5: 330DC3B84B961FAC517B5F92875FD051
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| Jezik: | Angleški jezik |
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| Tipologija: | 1.01 - Izvirni znanstveni članek |
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| Organizacija: |  NIB - Nacionalni inštitut za biologijo
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| Povzetek: | 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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| Ključne besede: | digital PCR, dPCR, quantification, multiplexing, genetically modified organisms, 6-color system, virus diagnostics, virology |
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| Status publikacije: | Objavljeno |
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| Verzija publikacije: | Objavljena publikacija |
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| Datum objave: | 01.11.2023 |
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| Leto izida: | 2023 |
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| Št. strani: | str. [1]-17 |
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| Številčenje: | iss. 22, art. no. ǂ4156 |
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| PID: | 20.500.12556/DiRROS-18583  |
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| UDK: | 578 |
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| ISSN pri članku: | 2304-8158 |
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| DOI: | 10.3390/foods12224156 |
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| COBISS.SI-ID: | 173360131 |
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| Opomba: | 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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| Datum objave v DiRROS: | 29.03.2024 |
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| Število ogledov: | 118 |
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| Število prenosov: | 49 |
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| Metapodatki: |     |
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