Real-Time PCR Array as a Universal Platform for the Detection of Genetically Modified Crops and Its Application in Identifying Unapproved Genetically Modified Crops in Japan

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• 作者：Junichi Mano ; Natsuki Shigemitsu ; Satoshi Futo ; Hiroshi Akiyama ; Reiko Teshima ; Akihiro Hino ; Satoshi Furui ; Kazumi Kitta
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2009
• 出版时间：January 14, 2009
• 年：2009
• 卷：57
• 期：1
• 页码：26-37
• 全文大小：361K
• 年卷期：v.57,no.1(January 14, 2009)
• ISSN：1520-5118

文摘

We developed a novel type of real-time polymerase chain reaction (PCR) array with TaqMan chemistry as a platform for the comprehensive and semiquantitative detection of genetically modified (GM) crops. Thirty primer-probe sets for the specific detection of GM lines, recombinant DNA (r-DNA) segments, endogenous reference genes, and donor organisms were synthesized, and a 96-well PCR plate was prepared with a different primer-probe in each well as the real-time PCR array. The specificity and sensitivity of the array were evaluated. A comparative analysis with the data and publicly available information on GM crops approved in Japan allowed us to assume the possibility of unapproved GM crop contamination. Furthermore, we designed a Microsoft Excel spreadsheet application, Unapproved GMO Checker version 2.01, which helps process all the data of real-time PCR arrays for the easy assumption of unapproved GM crop contamination. The spreadsheet is available free of charge at http://cse.naro.affrc.go.jp/jmano/index.html.