| 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 - Source URL, visit https://doi.org/10.3390/foods12224156
 PDF - Source file, download (1,83 MB)
MD5: 330DC3B84B961FAC517B5F92875FD051
|
|---|
| Language: | English |
|---|
| Typology: | 1.01 - Original Scientific Article |
|---|
| Organization: |  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  |
|---|
| UDC: | 578 |
|---|
| ISSN on article: | 2304-8158 |
|---|
| DOI: | 10.3390/foods12224156 |
|---|
| COBISS.SI-ID: | 173360131 |
|---|
| 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: | 119 |
|---|
| Downloads: | 49 |
|---|
| Metadata: |     |
|---|
|
:
|
Copy citation |
|---|
| | | | Share: |  |
|---|
Hover the mouse pointer over a document title to show the abstract or click
on the title to get all document metadata. |
