title: Interlaboratory study of qualitative PCR methods for genetically modified maize events MON810, bt11, GA21, and CaMV P35S.
abstract: Qualitative PCR methods for the genetically modified (GM) maize events MON810, Bt11, and GA21, and the 35S promoter (P35S) region of the cauliflower mosaic virus (CaMV) were evaluated in an interlaboratory study. Real-time PCR-based quantitative methods for these GM events using the same primer pairs had already been validated in previous studies. Fifteen laboratories in Japan participated in this interlaboratory study. Each participant extracted DNA from blind samples, performed qualitative PCR assays, and then detected the PCR products with agarose gel electrophoresis. The specificity, sensitivity, and false-negative and false-positive rates of these methods were determined with different concentrations of GM mixing samples. LODs of these methods for MON810, Bt11, GA21, and the P35S segment calculated as the amount of MON810 were 0.2, 0.2, 0.1, and 0.2% or less, respectively, indicating that the LODs of MON810, Bt11, and P35S were lower than 10 copies, and the LOD of GA21 was lower than 25 copies of maize haploid genome. The current study demonstrated that the qualitative methods would be fit for the detection and identification of these GM maize events and the P35S segment.
authors.affiliation: National Agriculture and Food Research Organization, National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
authors.forename: Reona
authors.initials: R
authors.lastname: Takabatake
authors.forename: Kaori
authors.initials: K
authors.lastname: Takashima
authors.forename: Takeyo
authors.initials: T
authors.lastname: Kurashima
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
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Koiwa
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Akiyama
authors.forename: Reiko
authors.initials: R
authors.lastname: Teshima
authors.forename: Satoshi
authors.initials: S
authors.lastname: Futo
authors.forename: Yasutaka
authors.initials: Y
authors.lastname: Minegishi
mesh_terms: D018744:DNA, Plant / Q000235:genetics*; D030841:Food, Genetically Modified; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000379:methods*; D003313:Zea mays / Q000235:genetics*
chemical_list: D018744:DNA, Plant