Evaluation of Quantitative PCR Methods for Genetically Modified Maize (MON863, NK603, TC1507 and T25)

Abstract

Novel real-time PCR-based quantitative methods were developed for three GM maize events; MON863, NK603 and TC1507. The quantitative methods were designed to amplify an event-specific segment for MON863 and NK603, and a construct-specific segment for TC1507. We also developed an event-specific quantitative method for T25. The conversion factor (Cf), which is required for calculating the GMO amount, was determined using three types of real-time PCR equipment; the ABI PRISM 7700,7900HT and 7500. The quantitative methods were evaluated by blind testing in an interlaboratory study using the ABI PRISM 7700 and 7900HT, and in a multilaboratory trial using the ABI PRISM 7500. The trueness, precision, and limit of quantitation were determined. Although the biases expressing the trueness for MON863, TC1507, and T25 were slightly high, all the data suggested that the developed methods were suitable for identification and quantification of these GM maize events.