title: International ring trial for the validation of an event-specific Golden Rice 2 quantitative real-time polymerase chain reaction method.
abstract: This article describes the international validation of the quantitative real-time polymerase chain reaction (PCR) detection method for Golden Rice 2. The method consists of a taxon-specific assay amplifying a fragment of rice Phospholipase D α2 gene, and an event-specific assay designed on the 3' junction between transgenic insert and plant DNA. We validated the two assays independently, with absolute quantification, and in combination, with relative quantification, on DNA samples prepared in haploid genome equivalents. We assessed trueness, precision, efficiency, and linearity of the two assays, and the results demonstrate that both the assays independently assessed and the entire method fulfill European and international requirements for methods for genetically modified organism (GMO) testing, within the dynamic range tested. The homogeneity of the results of the collaborative trial between Europe and Asia is a good indicator of the robustness of the method.
authors.affiliation: †Molecular Biology and Genomics Unit, Institute for Health and Consumer Protection, European Commission Joint Research Centre, Via E. Fermi 2749, 21027 Ispra, Varese, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Jacchia
authors.affiliation: †Molecular Biology and Genomics Unit, Institute for Health and Consumer Protection, European Commission Joint Research Centre, Via E. Fermi 2749, 21027 Ispra, Varese, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Nardini
authors.affiliation: †Molecular Biology and Genomics Unit, Institute for Health and Consumer Protection, European Commission Joint Research Centre, Via E. Fermi 2749, 21027 Ispra, Varese, Italy.
authors.forename: Niccolò
authors.initials: N
authors.lastname: Bassani
authors.affiliation: †Molecular Biology and Genomics Unit, Institute for Health and Consumer Protection, European Commission Joint Research Centre, Via E. Fermi 2749, 21027 Ispra, Varese, Italy.
authors.forename: Christian
authors.initials: C
authors.lastname: Savini
authors.affiliation: ‡Plant Breeding, Genetics and Biotechnology Division, International Rice Research Institute, Los Baños, 4031 Laguna, Philippines.
authors.forename: Jung-Hyun
authors.initials: JH
authors.lastname: Shim
authors.affiliation: ‡Plant Breeding, Genetics and Biotechnology Division, International Rice Research Institute, Los Baños, 4031 Laguna, Philippines.
authors.forename: Kurniawan
authors.initials: K
authors.lastname: Trijatmiko
authors.affiliation: †Molecular Biology and Genomics Unit, Institute for Health and Consumer Protection, European Commission Joint Research Centre, Via E. Fermi 2749, 21027 Ispra, Varese, Italy.
authors.forename: Joachim
authors.initials: J
authors.lastname: Kreysa
authors.affiliation: †Molecular Biology and Genomics Unit, Institute for Health and Consumer Protection, European Commission Joint Research Centre, Via E. Fermi 2749, 21027 Ispra, Varese, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Mazzara
mesh_terms: D001208:Asia; D005060:Europe; D012275:Oryza / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D010739:Phospholipase D / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics; D030821:Plants, Genetically Modified / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards
chemical_list: D010940:Plant Proteins; D010739:Phospholipase D
keywords: Golden Rice 2 (GR2); endogenous taxon-specific gene; event-specific method; genetically modified organism (GMO); international validation; quantitative real-time PCR (qPCR)