The Study On The Detection Of Genetically Modified Soybean And Soybean Products Using Loop-mediated Isothermal Aplification

The transgenic technique makes the improvement of varieties, including the increase ofoutput, the enhancement of quality and resistant and so on. Since1980s, genetic engineeringmade a rapid development, and it brings more convenient for our life. Nowadays, many GMplants have appeared in our lives. While bringing the infinite profit to human beings, thesecurity of transgenic technique was still questioned by people. Since the transgenic techniqueemerged, people were all along thinking and arguing about its food safety, ecology security.Considering the GM food’s safety, relevant laws and regulations have been issued to label theGMOs and their derived products in many counties. It is necessary to strengthen the GMOdetection. Only when we detect transgenic plant exactly can we monitor the commercialtransgenic food effectively. And many countries have strict rule about transgenic content infoods, so it is a key process, especially during the imports and exports trade.This paper was established by DNA loop-mediated isothermal amplification (LAMP)method for detection of genetically modified soybean and soybean products. LAMP is a novelnucleic acid amplification method developed by Eiken Chemical Co., Ltd., Japan. It ischaracterized by the use of4different primers specifically designed to recognize6distinctregions on the target gene and the reaction process proceeds at a constant temperature using aDNA polymerase with strand displacement activity.Firstly, the specific LAMP primers of genetically Modified were designed on the basispublished sequence of lectin gene, CaMV35S promoter, NOS terminator and two GMsoybean strain GTS40-3-2and MON89788, with the LAMP primer design support softwareprogram, using primer explorer software. The reaction conditions were optimized. There weretwo methods to observe the reaction results: the real-time turbiditimeter and adding SYBIGreen I. Secondary, the CTAB method was selected to extraction DNA from soybeanproducts. And PCR and LAMP assays were implemented to verify the accuracy of theLAMP method. The results showed that the CTAB method can successfully extract DNAfrom soybean products. The PCR results are consistent with LAMP results, showing thatLAMP method has a good accuracy. Finally, the extraction efficiency of three differentextraction methods (CTAB, the kit, carrier precipitation) for soybean oil was compared.Then genetically modified soybean oil were detected by the established LAMP methods andcorresponding fluorescent PCR reactions. And it was found the third method_carrierprecipitation was the most efficiency method which can be successfully extracted DNA fromsoybean oil. Results of the LAMP reaction is consistent with the results of fluorescent PCR,further illustrated the accuracy of the LAMP method.The combination of DNA extraction and detection system of response and detectionprocess, the overall specificity and sensitivity of detection the EU standard have reached. Itestablished a sound technology platform to realize rapid detection of genetically modified,laid an important foundation for further development of the industry or national standards.