Alexandra Bogožalec Košir (Avtor), Sabine Muller (Avtor), Jana Žel (Avtor), Mojca Milavec (Avtor), Allison C. Mallory (Avtor), David Dobnik (Avtor)

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.

Ključne besede

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;

Podatki

Jezik: Angleški jezik
Leto izida:
Tipologija: 1.01 - Izvirni znanstveni članek
Organizacija: NIB - Nacionalni inštitut za biologijo
UDK: 578
COBISS: 173360131 Povezava se bo odprla v novem oknu
ISSN: 2304-8158
Št. ogledov: 166
Št. prenosov: 65
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Slovenski jezik
Sekundarne ključne besede: digitalna PCR;digitalna verižna reakcija s polimerazo;kvantifikacija;multipleksiranje;gensko spremenjeni organizmi;6-barvni sistem;diagnostika virusov;virologija;
Komentar vira: Soavtorji: Sabine Muller, Jana Žel, Mojca Milavec, Allison C. Mallory and David Dobnik; Nasl. z nasl. zaslona; Opis vira z dne 23. 11. 2023;
Strani: str. [1]-17
Letnik: ǂiss. ǂ22
Zvezek: ǂart. no. ǂ4156
Čas izdaje: Nov. 2023
DOI: 10.3390/foods12224156
ID: 23284779