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

被引:25
作者
Toyota, A
Akiyama, H
Sugimura, M
Watanabe, T
Kikuch, H
Kanamori, H
Hino, A
Esaka, M
Maitani, T
机构
[1] Natl Inst Hlth Sci, Setagaya Ku, Tokyo 1588501, Japan
[2] Hiroshima Prefectural Inst Publ Hlth & Environm, Minami Ku, Hiroshima 7340007, Japan
[3] Natl Food Res Inst, Tsukuba, Ibaraki 3058642, Japan
[4] Hiroshima Univ, Grad Sch Biosphere Sci, Hiroshima 7398528, Japan
关键词
genetically modified soybean; Roundup Ready (R) Soybean; capillary-type real-time PCR system;
D O I
10.1271/bbb.70.821
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 (R) Soybean (RRS) quantification using a combination of a capillary-type real-time PCR system, a LightCycler (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 (C-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 front 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.
引用
收藏
页码:821 / 827
页数:7
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