title: Interlaboratory validation of quantitative duplex real-time PCR method for screening analysis of genetically modified maize.
abstract: To reduce the cost and time required to routinely perform the genetically modified organism (GMO) test, we developed a duplex quantitative real-time PCR method for a screening analysis simultaneously targeting an event-specific segment for GA21 and Cauliflower Mosaic Virus 35S promoter (P35S) segment [Oguchi et al., J. Food Hyg. Soc. Japan, 50, 117-125 (2009)]. To confirm the validity of the method, an interlaboratory collaborative study was conducted. In the collaborative study, conversion factors (Cfs), which are required to calculate the GMO amount (%), were first determined for two real-time PCR instruments, the ABI PRISM 7900HT and the ABI PRISM 7500. A blind test was then conducted. The limit of quantitation for both GA21 and P35S was estimated to be 0.5% or less. The trueness and precision were evaluated as the bias and reproducibility of the relative standard deviation (RSD(R)). The determined bias and RSD(R) were each less than 25%. We believe the developed method would be useful for the practical screening analysis of GM maize.
authors.affiliation: Analytical Science Division, National Food Research Institute, National Agriculture and Food Research Organization, Ibaraki.
authors.forename: Reona
authors.initials: R
authors.lastname: Takabatake
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Koiwa
authors.forename: Masaki
authors.initials: M
authors.lastname: Kasahara
authors.forename: Kaori
authors.initials: K
authors.lastname: Takashima
authors.forename: Satoshi
authors.initials: S
authors.lastname: Futo
authors.forename: Yasutaka
authors.initials: Y
authors.lastname: Minegishi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Akiyama
authors.forename: Reiko
authors.initials: R
authors.lastname: Teshima
authors.forename: Taichi
authors.initials: T
authors.lastname: Oguchi
authors.forename: Junichi
authors.initials: J
authors.lastname: Mano
authors.forename: Satoshi
authors.initials: S
authors.lastname: Furui
authors.forename: Kazumi
authors.initials: K
authors.lastname: Kitta
mesh_terms: D017796:Caulimovirus / Q000235:genetics; D017931:DNA Primers; D018744:DNA, Plant / Q000032:analysis; D030821:Plants, Genetically Modified / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D003313:Zea mays / Q000235:genetics*
chemical_list: D017931:DNA Primers; D018744:DNA, Plant