Portal de Periódicos da CAPES

Comprehensive GMO Detection Using Real-Time PCR Array: Single-Laboratory Validation

2012; Oxford University Press; Volume: 95; Issue: 2 Linguagem: Inglês

10.5740/jaoacint.11-388

1944-7922

Junichi Mano, Mioko Harada, Reona Takabatake, Satoshi Furui, Kazumi Kitta, Kôsuke Nakamura, Hiroshi Akiyama, Reiko Teshima, Hiromichi Noritake, Shuko Hatano, Satoshi Futo, Yasutaka Minegishi, Tayoshi Iizuka,

Insect Resistance and Genetics

Abstract 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.