title: [Development and validation of event-specific quantitative PCR method for genetically modified maize LY038].
abstract: In this article, we report a novel real-time PCR-based analytical method for quantitation of the GM maize event LY038. We designed LY038-specific and maize endogenous reference DNA-specific PCR amplifications. After confirming the specificity and linearity of the LY038-specific PCR amplification, we determined the conversion factor required to calculate the weight-based content of GM organism (GMO) in a multilaboratory evaluation. Finally, in order to validate the developed method, an interlaboratory collaborative trial according to the internationally harmonized guidelines was performed with blind DNA samples containing LY038 at the mixing levels of 0, 0.5, 1.0, 5.0 and 10.0%. The precision of the method was evaluated as the RSD of reproducibility (RSDR), and the values obtained were all less than 25%. The limit of quantitation of the method was judged to be 0.5% based on the definition of ISO 24276 guideline. The results from the collaborative trial suggested that the developed quantitative method would be suitable for practical testing of LY038 maize.
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization, Kannondai, Tsukuba, Japan.
authors.forename: Junichi
authors.initials: J
authors.lastname: Mano
authors.forename: Tomoko
authors.initials: T
authors.lastname: Masubuchi
authors.forename: Shuko
authors.initials: S
authors.lastname: Hatano
authors.forename: Satoshi
authors.initials: S
authors.lastname: Futo
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Koiwa
authors.forename: Yasutaka
authors.initials: Y
authors.lastname: Minegishi
authors.forename: Akio
authors.initials: A
authors.lastname: Noguchi
authors.forename: Kazunari
authors.initials: K
authors.lastname: Kondo
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Akiyama
authors.forename: Reiko
authors.initials: R
authors.lastname: Teshima
authors.forename: Takeyo
authors.initials: T
authors.lastname: Kurashima
authors.forename: Reona
authors.initials: R
authors.lastname: Takabatake
authors.forename: Kazumi
authors.initials: K
authors.lastname: Kitta
mesh_terms: D018744:DNA, Plant / Q000032:analysis* / Q000302:isolation & purification; D005504:Food Analysis / Q000379:methods*; D005515:Food Labeling / Q000592:standards; D030841:Food, Genetically Modified / Q000592:standards; D059021:Laboratory Proficiency Testing; D030821:Plants, Genetically Modified / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D003313:Zea mays / Q000235:genetics*
chemical_list: D018744:DNA, Plant