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

摘要

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.