Development and Validation of an Event-Specific Quantitative PCR Method for Genetically Modified Maize MIR162

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  • TAKABATAKE Reona
    Analytical Science Division, National Food Research Institute, National Agriculture and Food Research Organization
  • MASUBUCHI Tomoko
    Analytical Science Division, National Food Research Institute, National Agriculture and Food Research Organization
  • FUTO Satoshi
    FASMAC Co., Ltd.
  • MINEGISHI Yasutaka
    NIPPON GENE Co., Ltd.
  • NOGUCHI Akio
    National Institute of Health Sciences
  • KONDO Kazunari
    National Institute of Health Sciences
  • TESHIMA Reiko
    National Institute of Health Sciences
  • KURASHIMA Takeyo
    Analytical Science Division, National Food Research Institute, National Agriculture and Food Research Organization
  • MANO Junichi
    Analytical Science Division, National Food Research Institute, National Agriculture and Food Research Organization
  • KITTA Kazumi
    Analytical Science Division, National Food Research Institute, National Agriculture and Food Research Organization

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  • 遺伝子組換えトウモロコシMIR162の系統特異的定量検知法の開発および妥当性確認

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Abstract

A novel real-time PCR-based analytical method was developed for the event-specific quantification of a genetically modified (GM) maize event, MIR162. We first prepared a standard plasmid for MIR162 quantification. The conversion factor (Cf) required to calculate the genetically modified organism (GMO) amount was empirically determined for two real-time PCR instruments, the Applied Biosystems 7900HT (ABI7900) and the Applied Biosystems 7500 (ABI7500) for which the determined Cf values were 0.697 and 0.635, respectively. To validate the developed method, a blind test was carried out in an interlaboratory study. The trueness and precision were evaluated as the bias and reproducibility of relative standard deviation (RSDr). The determined biases were less than 25% and the RSDr values were less than 20% at all evaluated concentrations. These results suggested that the limit of quantitation of the method was 0.5%, and that the developed method would thus be suitable for practical analyses for the detection and quantification of MIR162.

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