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Title:Fast and accurate multiplex identification and quantification of seven genetically modified soybean lines using six-color digital PCR
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)
Files:URL URL - Source URL, visit https://doi.org/10.3390/foods12224156
 
.pdf PDF - Presentation file, download (1,83 MB)
MD5: 330DC3B84B961FAC517B5F92875FD051
 
Language:English
Typology:1.01 - Original Scientific Article
Organization:Logo NIB - National Institute of Biology
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.
Keywords:digital PCR, dPCR, quantification, multiplexing, genetically modified organisms, 6-color system, virus diagnostics, virology
Publication status:Published
Publication version:Version of Record
Publication date:01.11.2023
Year of publishing:2023
Number of pages:str. [1]-17
Numbering:iss. 22, art. no. ǂ4156
PID:20.500.12556/DiRROS-18583 New window
UDC:578
ISSN on article:2304-8158
DOI:10.3390/foods12224156 New window
COBISS.SI-ID:173360131 New window
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;
Publication date in DiRROS:29.03.2024
Views:809
Downloads:321
Metadata:XML DC-XML DC-RDF
:
BOGOŽALEC KOŠIR, Alexandra, MULLER, Sabine, ŽEL, Jana, MILAVEC, Mojca, MALLORY, Allison C. and DOBNIK, David, 2023, Fast and accurate multiplex identification and quantification of seven genetically modified soybean lines using six-color digital PCR. Foods [online]. 2023. Vol. 22, no. 4156, p. 1–17. [Accessed 6 April 2025]. DOI 10.3390/foods12224156. Retrieved from: https://dirros.openscience.si/IzpisGradiva.php?lang=eng&id=18583
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Record is a part of a journal

Title:Foods
Shortened title:Foods
Publisher:MDPI
ISSN:2304-8158
COBISS.SI-ID:512252472 New window

Document is financed by a project

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:P4-0165-2022
Name:Biotehnologija in sistemska biologija rastlin
Funder:Other - Other funder or multiple funders
Funding programme:European Union, European Regional Development Fund

Licences

License:CC BY 4.0, Creative Commons Attribution 4.0 International
Link:http://creativecommons.org/licenses/by/4.0/
Description:This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

Language:Slovenian
Keywords:digitalna PCR, digitalna verižna reakcija s polimerazo, kvantifikacija, multipleksiranje, gensko spremenjeni organizmi, 6-barvni sistem, diagnostika virusov, virologija


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