Quantitative detection system for maize sample containing combined-trait genetically modified maize.
Various countries have established regulations that stipulate the labeling of agricultural commodities, feed, and food products that contain or are made from genetically modified (GM) material or that contain adventitious GM material in amounts that exceed certain threshold levels. While regulations in some countries refer to GM material on a weight per weight (w/w) percentage, the currently applied detection methods do not directly measure the w/w percentage of the GM material. Depending on the particular method and the sample matrix it is applied to, the conversion of analytical results to a w/w percentage is challenging or not possible. The first rapid PCR system for GM maize detection on a single kernel basis has been developed. The equipment for the grinding of individual kernels and a silica membrane-based 96-well DNA extraction kit were both significantly revised and optimized for this particular purpose, respectively. We developed a multiplex real-time PCR method for the rapid quantification of GM DNA sequences in the obtained DNA solutions. In addition, a multiplex qualitative PCR detection method allows for the simultaneous detection of different GM maize traits in each kernel and thereby for identification of individual kernels that contain a combination of two or more GM traits. Especially for grain samples that potentially contain combined-trait GM maize kernels, the proposed methods can deliver informative results in a rapid, precise, and reliable manner.
Akiyama
Hiroshi
H
National Institute of Health Sciences, Kamiyoga, Setagaya-ku, Tokyo, Japan. akiyama@nihs.go.jp
Watanabe
Takahiro
T
Wakabayashi
Kaoru
K
Nakade
Shinsuke
S
Yasui
Shuji
S
Sakata
Kozue
K
Chiba
Ryoko
R
Spiegelhalter
Frank
F
Hino
Akihiro
A
Maitani
Tamio
T
D018744:DNA, Plant / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism; D018745:Genome, Plant / Q000235:genetics; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D013997:Time Factors; D003313:Zea mays / Q000235:genetics*
D018744:DNA, Plant; D003851:Deoxyribonucleases