Determination of unprocessed genetically modified soybean in foods using simplex and duplex real-time PCR with an internal standard

摘要

We developed a method using an internal standard to determine the amount of unprocessed genetically modified organisms (GMO) in foods as GMO weight per weight of total food (w/w_(tf)) using real-time PCR. Food samples were spiked with an internal standard, a ColEl plasmid, and DNA was extracted from the samples using a silica membrane-type kit and analysed using real-time PCR. The relationship between the GMO amount and the copy number ratio of the transgene to ColEl in 0.1-5% w/w_(tf) GM soybean powders was found to have a correlation coefficient (r) of 0.97. GMO was quantified in food samples spiked with GM soybean (final amount 0.5% w/w_(tf) GMO). The average GMO amount ranged from 0.35% to 0.63% w/w_(tf). The results show that our method should be useful for quantifying unprocessed GMO in foods. We also developed a duplex assay, which is simpler and more accurate than the simplex assay.