Mano Junichi, Harada Mioko, Takabatake Reona, Furui Satoshi, Kitta Kazumi, Nakamura Kosuke, Akiyama Hiroshi, Teshima Reiko, Noritake Hiromichi, Hatano Shuko, Futo Satoshi, Minegishi Yasutaka, Iizuka Tayoshi
National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
J AOAC Int. 2012 Mar-Apr;95(2):508-16. doi: 10.5740/jaoacint.11-388.
We have developed a real-time PCR array method to comprehensively detect genetically modified (GM) organisms. In the method, genomic DNA extracted from an agricultural product is analyzed using various qualitative real-time PCR assays on a 96-well PCR plate, targeting for individual GM events, recombinant DNA (r-DNA) segments, taxon-specific DNAs, and donor organisms of the respective r-DNAs. In this article, we report the single-laboratory validation of both DNA extraction methods and component PCR assays constituting the real-time PCR array. We selected some DNA extraction methods for specified plant matrixes, i.e., maize flour, soybean flour, and ground canola seeds, then evaluated the DNA quantity, DNA fragmentation, and PCR inhibition of the resultant DNA extracts. For the component PCR assays, we evaluated the specificity and LOD. All DNA extraction methods and component PCR assays satisfied the criteria set on the basis of previous reports.