Interlaboratory validation of quantitative duplex real-time PCR method for screening analysis of genetically modified maize |
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Authors: | Takabatake Reona Koiwa Tomohiro Kasahara Masaki Takashima Kaori Futo Satoshi Minegishi Yasutaka Akiyama Hiroshi Teshima Reiko Oguchi Taichi Mano Junichi Furui Satoshi Kitta Kazumi |
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Affiliation: | Analytical Science Division, National Food Research Institute, National Agriculture and Food Research Organization, Ibaraki. |
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Abstract: | To reduce the cost and time required to routinely perform the genetically modified organism (GMO) test, we developed a duplex quantitative real-time PCR method for a screening analysis simultaneously targeting an event-specific segment for GA21 and Cauliflower Mosaic Virus 35S promoter (P35S) segment [Oguchi et al., J. Food Hyg. Soc. Japan, 50, 117-125 (2009)]. To confirm the validity of the method, an interlaboratory collaborative study was conducted. In the collaborative study, conversion factors (Cfs), which are required to calculate the GMO amount (%), were first determined for two real-time PCR instruments, the ABI PRISM 7900HT and the ABI PRISM 7500. A blind test was then conducted. The limit of quantitation for both GA21 and P35S was estimated to be 0.5% or less. The trueness and precision were evaluated as the bias and reproducibility of the relative standard deviation (RSD(R)). The determined bias and RSD(R) were each less than 25%. We believe the developed method would be useful for the practical screening analysis of GM maize. |
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