Establishment Of Multi-Fluorescence Real-Time Pcr Assay For Gm Soybean Detection

In the past two decades, a growing number of genetically modified (GM) crops have been developed and commercialized worldwide. High-throughput methods for screening and identification of GM crops have become a requirement for food industry and government agencies. To effectively detect seven specific transgenic soybean lines named GTS40-3-2, MON89788, CV127, A5547-127, A2704-12, 305423, and 356043, seven different primer-probe sets (labeled with FAM, HEX, Taxas Red, Cy5, and Quasar 705) were designed for the multi-fluorescence real-time PCR (MF-RT-PCR) assay. Through a series of experiments and tests including of primer combination screening, system optimization, specificity test, sensitivity test, and applicability test, two GM soybean target specific systems, SOY-M1 and SOY-M2, were established. SOY-M1, a pentaplex PCR detection system, could efficiently identify Mon89788, A2704-12, 305423, and 356043, and SOY-M2, a tetraplex PCR detection system is able to detect GTS40-3-2 CV127, and A5547-127. The sensitivity of both systems is 0.1%. In summary, the MF-RT-PCR detection system established in this study is highly accurate, reliable, and efficient in detecting these seven soybean lines and their derivates. (C) 2020 Friends Science Publishers