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
digitalna PCR;digitalna verižna reakcija s polimerazo;kvantifikacija;multipleksiranje;gensko spremenjeni organizmi;6-barvni sistem;diagnostika virusov;virologija;digital PCR;dPCR;quantification;multiplexing;genetically modified organisms;6-color system;virus diagnostics;virology;
Data
Language: |
English |
Year of publishing: |
2023 |
Typology: |
1.01 - Original Scientific Article |
Organization: |
NIB - National Institute of Biology |
UDC: |
578 |
COBISS: |
173360131
|
ISSN: |
2304-8158 |
Views: |
166 |
Downloads: |
65 |
Average score: |
0 (0 votes) |
Metadata: |
|
Other data
Secondary language: |
Slovenian |
Secondary keywords: |
digitalna PCR;digitalna verižna reakcija s polimerazo;kvantifikacija;multipleksiranje;gensko spremenjeni organizmi;6-barvni sistem;diagnostika virusov;virologija; |
Source comment: |
Soavtorji: Sabine Muller, Jana Žel, Mojca Milavec, Allison C. Mallory and David Dobnik;
Nasl. z nasl. zaslona;
Opis vira z dne 23. 11. 2023;
|
Pages: |
str. [1]-17 |
Volume: |
ǂiss. ǂ22 |
Issue: |
ǂart. no. ǂ4156 |
Chronology: |
Nov. 2023 |
DOI: |
10.3390/foods12224156 |
ID: |
23284779 |