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Quantification of genetically modified soybeans using a combination of a capillary-type real-time PCR system and a plasmid reference standard
Authors:Toyota Akie  Akiyama Hiroshi  Sugimura Mitsunori  Watanabe Takahiro  Kikuchi Hiroyuki  Kanamori Hisayuki  Hino Akihiro  Esaka Muneharu  Maitani Tamio
Affiliation:Hiroshima Prefectural Institute of Public Health and Environment, Minami-ku, Hiroshima, Japan.
Abstract:Because the labeling of grains and feed- and foodstuffs is mandatory if the genetically modified organism (GMO) content exceeds a certain level of approved genetically modified varieties in many countries, there is a need for a rapid and useful method of GMO quantification in food samples. In this study, a rapid detection system was developed for Roundup Ready Soybean (RRS) quantification using a combination of a capillary-type real-time PCR system, a LightCycler real-time PCR system, and plasmid DNA as the reference standard. In addition, we showed for the first time that the plasmid and genomic DNA should be similar in the established detection system because the PCR efficiencies of using plasmid DNA and using genomic DNA were not significantly different. The conversion factor (Cf) to calculate RRS content (%) was further determined from the average value analyzed in three laboratories. The accuracy and reproducibility of this system for RRS quantification at a level of 5.0% were within a range from 4.46 to 5.07% for RRS content and within a range from 2.0% to 7.0% for the relative standard deviation (RSD) value, respectively. This system rapidly monitored the labeling system and had allowable levels of accuracy and precision.
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