Validation of real-time PCR analyses for line-specific quantitation of genetically modified maize and soybean using new reference molecules.
Validation of real-time PCR analyses for line-specific quantitation of genetically modified maize and soybean using new reference molecules.
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使用新参考分子验证转基因玉米和大豆品系特异性定量的实时 PCR 分析。
DOI：
10.1093/jaoac/85.5.1119
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发表时间：
2002
影响因子：
1.6
通讯作者：
A. Hino
中科院分区：
文献类型：
--
作者：
Yoichiro Shindo;H. Kuribara;T. Matsuoka;S. Futo;C. Sawada;Jinji Shono;H. Akiyama;Y. Goda;M. Toyoda;A. Hino
基于使用新的参考分子的实时定量聚合酶链式反应的新分析方法在实验室间的转基因玉米和大豆的定量研究中得到验证。来自日本、韩国和美国的13个实验室参与了这项研究。实验室间的研究包括两个不同的阶段：(1)转基因玉米和转基因大豆种子中系数值、重组DNA(r-DNA)序列和内源DNA序列比率的测定试验；(2)6对玉米和大豆样品的盲法试验，包括不同水平的转基因玉米和转基因大豆。试验结果表明，该方法适用于5个转基因玉米品系和1个转基因大豆品系的特定定量。经统计处理剔除异常值后，这些方法在5.0%水平上的重复性和重复性分别为13.7%和15.9%。方法对BT11、T25和MON810的定量限为0.50%，对GA21、Event176和Roundup Ready大豆的定量限为0.10%。盲测结果表明，这些方法所获得的数值信息将有助于转基因作物标签系统的实际分析。
Novel analytical methods based on real-time quantitative polymerase chain reactions by use of new reference molecules were validated in interlaboratory studies for the quantitation of genetically modified (GM) maize and soy. More than 13 laboratories from Japan, Korea, and the United States participated in the studies. The interlaboratory studies included 2 separate stages: (1) measurement tests of coefficient values, the ratio of recombinant DNA (r-DNA) sequence, and endogenous DNA sequence in the seeds of GM maize and GM soy; and (2) blind tests with 6 pairs of maize and soy samples, including different levels of GM maize or GM soy. Test results showed that the methods are applicable to the specific quantitation of the 5 lines of GM maize and one line of GM soy. After statistical treatment to remove outliers, the repeatability and reproducibility of these methods at a level of 5.0% were <13.7 and 15.9%, respectively. The quantitation limits of the methods were 0.50% for Bt11, T25, and MON810, and 0.10% for GA21, Event176, and Roundup Ready soy. The results of blind tests showed that the numerical information obtained from these methods will contribute to practical analyses for labeling systems of GM crops.