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.
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 .
Mano
Junichi
J
National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki, Japan.
Shigemitsu
Natsuki
N
Futo
Satoshi
S
Akiyama
Hiroshi
H
Teshima
Reiko
R
Hino
Akihiro
A
Furui
Satoshi
S
Kitta
Kazumi
K
D018556:Crops, Agricultural / Q000235:genetics*; D018744:DNA, Plant / Q000032:analysis; D004274:DNA, Recombinant / Q000032:analysis; D002523:Edible Grain / Q000235:genetics; D005506:Food Contamination / Q000032:analysis; D007564:Japan; D007881:Legislation, Food; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D018744:DNA, Plant; D004274:DNA, Recombinant