title: Development of a multiplex polymerase chain reaction method for simultaneous detection of eight events of genetically modified maize.
abstract: In this study, we developed a novel multiplex polymerase chain reaction (PCR) method for simultaneous detection of up to eight events of genetically modified (GM) maize within a single reaction. The eight detection primer pairs designed to be construct specific for eight respective GM events (i.e., Bt11, Event176, GA21, MON810, MON863, NK603, T25, and TC1507) and a primer pair for an endogenous reference gene, ssIIb, were included in the nonaplex(9plex) PCR system, and its amplified products could be distinguished by agarose gel and capillary electrophoreses based on their different lengths. The optimal condition enabled us to reliably amplify two fragments corresponding to a construct specific sequence and a taxon specific ssIIb in each of the eight events of GM maize and all of nine fragments in a simulated GM mixture containing as little as 0.25% (w/w) each of eight events of GM maize. These results indicate that this multiplex PCR method could be an effective qualitative detection method for screening GM maize.
authors.affiliation: Research and Development Division, Fasmac Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Onishi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Matsuoka
authors.forename: Takashi
authors.initials: T
authors.lastname: Kodama
authors.forename: Koichi
authors.initials: K
authors.lastname: Kashiwaba
authors.forename: Satoshi
authors.initials: S
authors.lastname: Futo
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Akiyama
authors.forename: Tamio
authors.initials: T
authors.lastname: Maitani
authors.forename: Satoshi
authors.initials: S
authors.lastname: Furui
authors.forename: Taichi
authors.initials: T
authors.lastname: Oguchi
authors.forename: Akihiro
authors.initials: A
authors.lastname: Hino
mesh_terms: D001483:Base Sequence; D018744:DNA, Plant; D004587:Electrophoresis, Agar Gel; D019075:Electrophoresis, Capillary; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D003313:Zea mays / Q000235:genetics*
chemical_list: D018744:DNA, Plant