Digital PCR as an effective tool for GMO quantification in complex matrices

Alexandra Bogožalec Košir (Author), Tina Demšar (Author), Dejan Štebih (Author), Jana Žel (Author), Mojca Milavec (Author)

Abstract

The increased use of genetically modified organisms (GMOs) is accompanied by increased complexity of the matrices that contain GMOs. The most common DNA-based approach for GMO detection and quantification is real-time quantitative polymerase chain reaction (qPCR). However, as qPCR is sensitive to inhibitors and relies on standard curves for quantification, it has limited application in GMO quantification for complex matrices. To overcome this hurdle in DNA quantification, we present droplet digital PCR (ddPCR) assays that were designed to target ‘Roundup Ready’ soybean and the soybean reference gene. Three ddPCR assays were transferred from qPCR to QX100/QX200 ddPCR platforms and characterised. Together, the fitness-for-purpose study on four real-life samples and the use of a chamber-based PCR system, showed that dPCR has great potential to improve such measurements in GMO testing and monitoring of food authenticity.

Keywords

genetically modified organisms;digital PCR;GMO quantification;complex matrices;

Data

Language:	English
Year of publishing:	2019
Typology:	1.01 - Original Scientific Article
Organization:	NIB - National Institute of Biology
UDC:	577.2
COBISS:	5062223
ISSN:	0308-8146
Views:	32
Downloads:	6
Average score:	0 (0 votes)
Metadata:

Other data

Pages:	str. 73-78
Issue:	ǂVol. ǂ294
Chronology:	2019
DOI:	10.1016/j.foodchem.2019.05.029
ID:	24579924