Development of Multiplex PCR Method for Simultaneous Detection of Four Events of Genetically Modified Maize: DAS-59122-7, MIR604, MON863 and MON88017

Abstract

A novel multiplex PCR method was developed for simultaneous event-specific detection of four events of GM maize, i.e., DAS-59122-7, MIR604, MON88017, and MON863. The single laboratory examination of analytical performance using simulated DNA mixtures containing GM DNA at various concentrations in non-GM DNA suggested that the limits of detection (LOD) of the multiplex PCR method were 0.16ῌ for MON863, MIR604, and MON88017, and 0.078ῌ for DAS-59122-7. We previously developed a nonaplex (9plex) PCR method for eight events of GM maize, i.e., Bt11, Bt176, GA21, MON810, MON863, NK603, T25, and TC1507. Together with the nonaplex PCR method, the newly developed method enabled the detection and identification of eleven GM maize events that are frequently included in commercial GM seed used in Japan. In addition, this combinational analysis may be useful for the identification of combined event products of GM maize.

Application of polymerase chain reaction with disposable amplicon detection device for identification of regulatory gene introduced into genetically modified maize

“…We also added a primer pair specific to the SSIIb gene as an experimental control for amplification. The SSIIb gene is a single copy gene in the maize genome [Oguchi et al, 2010].…”

A novel trait-specific real-time PCR method enables quantification of genetically modified (GM) maize content in ground grain samples containing stacked GM maize

“…Furthermore, the corrected C f for cp4epsps or pat would need to be consecutively updated by monitoring the GM maize events in non-IP maize samples imported into Japan, because the rates of the GM maize events would be predicted to vary every year. In addition, the trait-specific method can prove GMO content, but not the existence of stacked GM maize events in maize sample, whereas the individual kernel detection method enables identification of stacked GM maize event in a maize kernel by combining with the event-specific method [17,18,44].…”

“…The GMO content of maize samples containing stacked GM maize is generally overestimated when determined using qPCR methods, as compared to the actual w/w percentage of GM maize, because a kernel of stacked GM maize contains the GMspecific DNAs in proportion to the number of GM events. To avoid overestimation, we developed an individual kernel detection method that involves multiplex real-time PCR using the extracted DNA from individual ground maize kernels [15][16][17][18]. This detection system has already been implemented in Japan as an official GM maize detection method [19].…”

Composition and rheological properties of flour and dough from genetically modified wheat (Triticum aestivum L.) Hi-Line 111

“…Consumers have demanded appropriate information and labeling for foods derived from GM crops. Thus, various labeling systems have been introduced for GM foods (Oguchi et al 2010). Pressure from consumers and public demand has led many countries to require labeling for the presence of GMOs in foods and sensitive detection methods for GM foods and feeds (Yoke-Kqueen et al 2011;Ramadan and Elsanhoty 2012;Elsanhoty et al 2013;Castigliego et al 2015;Turkec et al 2016).…”