Development and Evaluation of Event-Specific Qualitative PCR Methods for Genetically Modified Bt10 Maize

• Published in: Journal of agricultural and food chemistry Vol. 55; no. 4; pp. 1274 - 1279
• Main Authors: Watanabe, Takahiro, Tokishita, Shoko, Spiegelhalter, Frank, Furui, Satoshi, Kitta, Kazumi, Hino, Akihiro, Matsuda, Rieko, Akiyama, Hiroshi, Maitani, Tamio
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 21.02.2007
• Subjects: Biological and medical sciences; Bt10 corn; Cereal and baking product industries; corn; food analysis; Food Contamination - analysis; Food industries; Fundamental and applied biological sciences. Psychology; genetically modified foods; Plants, Genetically Modified - genetics; polymerase chain reaction; Polymerase Chain Reaction - methods; qualitative analysis; qualitative polymerase chain reaction; recombinant DNA; Sensitivity and Specificity; Zea mays; Zea mays - classification; Zea mays - genetics; Evaluation; Monocotyledones; Zea mays; Corn; Method; Polymerase chain reaction; Gramineae; Angiospermae; Development; Cereal; Spermatophyta; Molecular biology; Genetically modified organism
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf062818v

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.