title: A high-throughput method for GMO multi-detection using a microfluidic dynamic array.
abstract: The ever-increasing production of genetically modified crops generates a demand for high-throughput DNA-based methods for the enforcement of genetically modified organisms (GMO) labelling requirements. The application of standard real-time PCR will become increasingly costly with the growth of the number of GMOs that is potentially present in an individual sample. The present work presents the results of an innovative approach in genetically modified crops analysis by DNA based methods, which is the use of a microfluidic dynamic array as a high throughput multi-detection system. In order to evaluate the system, six test samples with an increasing degree of complexity were prepared, preamplified and subsequently analysed in the Fluidigm system. Twenty-eight assays targeting different DNA elements, GM events and species-specific reference genes were used in the experiment. The large majority of the assays tested presented expected results. The power of low level detection was assessed and elements present at concentrations as low as 0.06 % were successfully detected. The approach proposed in this work presents the Fluidigm system as a suitable and promising platform for GMO multi-detection.
authors.affiliation: RIKILT Wageningen UR, P.O. Box 230, 6700 AE, Wageningen, The Netherlands.
authors.forename: Fábio Cristiano Angonesi
authors.initials: FC
authors.lastname: Brod
authors.forename: Jeroen P
authors.initials: JP
authors.lastname: van Dijk
authors.forename: Marleen M
authors.initials: MM
authors.lastname: Voorhuijzen
authors.forename: Andréia Zilio
authors.initials: AZ
authors.lastname: Dinon
authors.forename: Luis Henrique S
authors.initials: LH
authors.lastname: Guimarães
authors.forename: Ingrid M J
authors.initials: IM
authors.lastname: Scholtens
authors.forename: Ana Carolina Maisonnave
authors.initials: AC
authors.lastname: Arisi
authors.forename: Esther J
authors.initials: EJ
authors.lastname: Kok
mesh_terms: D027461:Beta vulgaris / Q000235:genetics*; D002138:Calibration; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification*; D057166:High-Throughput Screening Assays; D006801:Humans; D057230:Limit of Detection; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D013025:Glycine max / Q000235:genetics*; D003313:Zea mays / Q000235:genetics*
chemical_list: D017931:DNA Primers; D018744:DNA, Plant