20.500.12556/DiRROS-18583 Fast and accurate multiplex identification and quantification of seven genetically modified soybean lines using six-color digital PCR 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. digital PCR dPCR quantification multiplexing genetically modified organisms 6-color system virus diagnostics virology digitalna PCR digitalna verižna reakcija s polimerazo kvantifikacija multipleksiranje gensko spremenjeni organizmi 6-barvni sistem diagnostika virusov virologija true false true Angleški jezik Slovenski jezik Neznano 2024-03-29 11:42:31 2024-03-29 11:42:32 2024-05-23 11:45:26 0000-00-00 00:00:00 2023 0 0 Soavtorji: Sabine Muller, Jana Žel, Mojca Milavec, Allison C. Mallory and David Dobnik; Nasl. z nasl. zaslona; Opis vira z dne 23. 11. 2023; str. [1]-17 art. no. ǂ4156 iss. 22 Nov. 2023 0000-00-00 Zaloznikova Objavljeno apc 0000-00-00 0000-00-00 2023-11-01 578 2304-8158 10.3390/foods12224156 173360131 https://doi.org/10.3390/foods12224156 1 aad91238-edb0-11ee-9dd4-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=25342 RAZ_Bogozalec_Kosir_Alexandra_2023.pdf RAZ_Bogozalec_Kosir_Alexandra_2023.pdf 1 330DC3B84B961FAC517B5F92875FD051 9c824a004e6cf20896ad9445cc3228b7a6f76246e692f9b2f8a74fdbcb429d5b 239f3f8d-edb1-11ee-9dd4-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=25343 Nacionalni inštitut za biologijo 0 0 0