Quantification of genetically modified soybeans [Glycine max] using a combination of a capillary-type real-time PCR system and a plasmid reference standard

• Published in: Bioscience, biotechnology, and biochemistry Vol. 70; no. 4; pp. 821 - 827
• Main Authors: Toyota, A.(Hiroshima-ken. Inst. of Public Health and Environmental Sciences, Hiroshima (Japan)), Akiyama, H, Sugimura, M, Watanabe, T, Kikuchi, H, Kanamori, H, Hino, A, Esaka, M, Maitani, T
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 2006; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: ADN RECOMBINADO; ADN RECOMBINE; Biological and medical sciences; capillary-type real-time PCR system; CONTROL DEL ETIQUETADO; CONTROLE DU MARQUAGE; DIAGNOSIS; DIAGNOSTIC; DIAGNOSTICO; Food, Genetically Modified; Fundamental and applied biological sciences. Psychology; genetically modified soybean; Genome, Plant - genetics; GLYCINE MAX; Glycine max - genetics; LABELLING CONTROLS; PCR; PLANTAS TRANSGENICAS; PLANTE TRANSGENIQUE; Plants, Genetically Modified; PLASMIDE; PLASMIDIOS; PLASMIDS; Plasmids - genetics; Polymerase Chain Reaction - instrumentation; Polymerase Chain Reaction - methods; RECOMBINANT DNA; Reference Standards; Reproducibility of Results; Roundup Ready; Soybean; Time Factors; TRANSGENIC PLANTS; Soybean; Real time; Glycine max; Plasmid; Polymerase chain reaction; Leguminosae; Dicotyledones; Roundup Ready® Soybean; Angiospermae; genetically modified soybean; Spermatophyta; Genetically modified organism; Quantitative analysis; capillary-type real-time PCR system
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.70.821

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 Readysup(R) Soybean (RRS) quantification using a combination of a capillary-type real-time PCR system, a LightCyclersup(R) 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 (Csub(f)) 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.