Development and Evaluation of Event-Specific Quantitative PCR Method for Genetically Modified Soybean MON87701

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  • TSUKAHARA Keita
    Division of Analytical Science, Food Research Institute, National Agriculture and Food Research Organization FASMAC Co., Ltd.
  • TAKABATAKE Reona
    Division of Analytical Science, Food Research Institute, National Agriculture and Food Research Organization
  • MASUBUCHI Tomoko
    Division of Analytical Science, 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
  • NISHIMAKI-MOGAMI Tomoko
    National Institute of Health Sciences
  • KURASHIMA Takeyo
    Division of Analytical Science, Food Research Institute, National Agriculture and Food Research Organization
  • MANO Junichi
    Division of Analytical Science, Food Research Institute, National Agriculture and Food Research Organization
  • KITTA Kazumi
    Division of Analytical Science, Food Research Institute, National Agriculture and Food Research Organization

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  • 遺伝子組換えダイズMON87701の系統特異的定量PCR法の開発と性能評価

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Abstract

<p>A real-time PCR-based analytical method was developed for the event-specific quantification of a genetically modified (GM) soybean event, MON87701. First, a standard plasmid for MON87701 quantification was constructed. The conversion factor (Cf) required to calculate the amount of genetically modified organism (GMO) was experimentally determined for a real-time PCR instrument. The determined Cf for the real-time PCR instrument was 1.24. For the evaluation of the developed method, a blind test was carried out in an inter-laboratory trial. The trueness and precision were evaluated as the bias and reproducibility of relative standard deviation (RSDr), respectively. The determined biases and the RSDr values were less than 30 and 13%, respectively, at all evaluated concentrations. The limit of quantitation of the method was 0.5%, and the developed method would thus be applicable for practical analyses for the detection and quantification of MON87701.</p>

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