Development and interlaboratory validation of quantitative polymerase chain reaction method for screening analysis of genetically modified soybeans.
Development and interlaboratory validation of quantitative polymerase chain reaction method for screening analysis of genetically modified soybeans.
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用于转基因大豆筛选分析的定量聚合酶链反应方法的开发和实验室间验证。
DOI：
10.1248/bpb.b12-00766
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发表时间：
2013
影响因子：
2
通讯作者：
K. Kitta
中科院分区：
文献类型：
--
作者：
R. Takabatake;Mari Onishi;T. Koiwa;S. Futo;Y. Minegishi;H. Akiyama;R. Teshima;Takeyo Kurashima;J. Mano;S. Furui;K. Kitta
针对三种转基因大豆：RRS、A2704-12 和 MON89788，开发了一种基于实时聚合酶链反应 (PCR) 的新型定量筛选方法。花椰菜花叶病毒的 35S 启动子 (P35S) 被引入 RRS 和 A2704-12，但未引入 MON89788。然后，我们设计了一种将 P35S 定量和 MON89788 事件特异性定量相结合的筛选方法。通过实验确定将转基因生物(GMO)的量从拷贝数比转换为重量比所需的转换因子(Cf)。真实度和精密度分别通过相对标准偏差 (RSDR) 的偏差和再现性进行评估。该方法测定的两个目标的 RSDR 值均小于 25%。我们认为所开发的方法适合转基因生物的简单检测和近似定量。
A novel real-time polymerase chain reaction (PCR)-based quantitative screening method was developed for three genetically modified soybeans: RRS, A2704-12, and MON89788. The 35S promoter (P35S) of cauliflower mosaic virus is introduced into RRS and A2704-12 but not MON89788. We then designed a screening method comprised of the combination of the quantification of P35S and the event-specific quantification of MON89788. The conversion factor (Cf) required to convert the amount of a genetically modified organism (GMO) from a copy number ratio to a weight ratio was determined experimentally. The trueness and precision were evaluated as the bias and reproducibility of relative standard deviation (RSDR), respectively. The determined RSDR values for the method were less than 25% for both targets. We consider that the developed method would be suitable for the simple detection and approximate quantification of GMO.