Comprehensive GMO detection using real-time PCR array: single-laboratory validation.
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.
National Food Research Institute, National Agriculture and Food Research Organization, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Junichi
J
Mano
Mioko
M
Harada
Reona
R
Takabatake
Satoshi
S
Furui
Kazumi
K
Kitta
Kosuke
K
Nakamura
Hiroshi
H
Akiyama
Reiko
R
Teshima
Hiromichi
H
Noritake
Shuko
S
Hatano
Satoshi
S
Futo
Yasutaka
Y
Minegishi
Tayoshi
T
Iizuka
D001483:Base Sequence; D005504:Food Analysis / Q000379:methods*; D030781:Organisms, Genetically Modified / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D003313:Zea mays / Q000235:genetics*