Development and evaluation of event-specific qualitative PCR methods for genetically modified Bt10 maize.
Development and evaluation of event-specific qualitative PCR methods for genetically modified Bt10 maize.
复制标题
转基因 Bt10 玉米事件特异性定性 PCR 方法的开发和评估。
DOI：
10.1021/jf062818v
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发表时间：
2007
影响因子：
6.1
通讯作者：
T. Maitani
中科院分区：
文献类型：
--
作者：
Takahiro Watanabe;S. Tokishita;F. Spiegelhalter;S. Furui;K. Kitta;A. Hino;R. Matsuda;H. Akiyama;T. Maitani
2005 年，有报道称，被称为 Bt10 的转基因 (GM) 玉米品种或“事件”在过去 4 年里无意中在美国传播。为了确保用于食品和饲料生产的谷物不含有微量 Bt10 玉米并符合适用法规，需要对 Bt10 玉米进行高灵敏度和特异性检测。因此，我们开发了一种新型定性 PCR 系统，用于特异性检测 Bt10 玉米。此外，我们还充分评估了我们自己的和 Bt10 开发商先正达有限公司提供的两种 PCR 系统的性能特征。证实两种定性 PCR 系统都能特异性检测 Bt10 玉米，单实验室检查结果表明，两种方法的检出限均低于 0.05%。为了评估方法的重现性，我们组织了一项有 6 个实验室参与的实验室间研究，并对 240 个盲测样品进行了分析。在本文中，我们首次报告了从实验室间研究中获得的定性 PCR 数据的统计分析。分析结果还显示，上述两种方法的灵敏度没有显着差异，两种方法的检出限均小于0.05%。因此，我们得出结论，这两种方法同样适用于测试样品中未经批准的转基因玉米 Bt10 事件的正确识别和灵敏检测。
In 2005 it was reported that the genetically modified (GM) maize strain or "event" called Bt10 had been distributed inadvertently in the United States over the previous 4 years. In order to ensure that grain for food and feed production did not contain trace amounts of Bt10 maize and complied with the applicable regulation, highly sensitive and specific detection of Bt10 maize was required. Accordingly, we developed a novel qualitative PCR system for specific detection of Bt10 maize. Moreover, we amply evaluated the performance characteristics of two PCR systems, our own and the one provided by the developer of Bt10, Syngenta Co. Ltd. It was confirmed that both of the qualitative PCR systems can specifically detect Bt10 maize, and the results of a single-laboratory examination suggested that the limit of detection was approximately less than 0.05% for both methods. To evaluate the reproducibility of the methods, we organized an interlaboratory study with the participation of 6 laboratories and analysis of 240 blind test samples. In this paper, we report, for the first time, the statistical analysis of the qualitative PCR data obtained from the interlaboratory study. The results of this analysis also revealed that there was no significant difference in the sensitivity between the two aforementioned methods and that the limit of detection of both the methods was less than 0.05%. Thus, we conclude that both of the methods are equally suitable for correct identification and sensitive detection of the unapproved GM maize Bt10 event in test samples.