Rapid Detection of Genetically Modified Ingredients in Soybean Products by Real-Time Loop-Mediated Isothermal Amplification

Huiling Di¹, Lei Shi¹, Huiping Shen¹, He Yan¹, Hecheng Meng¹, Lili Li¹, M. Jahangir Alam², Shinji Yamasaki³ and Lei Ye^1,

¹College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, PR China

²University of Houston College of Pharmacy Houston, TX, USA

³Graduate Schools of Life and Environmental Sciences, Osaka Prefecture University, Osaka, Japan

Cite this paper:
Huiling Di, Lei Shi, Huiping Shen, He Yan, Hecheng Meng, Lili Li, M. Jahangir Alam, Shinji Yamasaki and Lei Ye. Rapid Detection of Genetically Modified Ingredients in Soybean Products by Real-Time Loop-Mediated Isothermal Amplification. Journal of Food and Nutrition Research. 2014; 2(7):363-368. doi: 10.12691/jfnr-2-7-6

Abstract

A real-time turbidimeter loop-mediated isothermal amplification (LAMP) assay was developed for the rapid, sensitive, and convenient detection of genetically modified (GM) ingredients in soybean products obtained from local supermarkets in China. Specific primers were designed based on the published sequence of lectin, nos terminator and endogenous genes of two GM soybean strains (GTS40-3-2 and MON89788). The total genomic DNA of soybean-made products was extracted by cetyltrimethylammonium bromide (CTAB) method. Then, the genomic DNA was used as template for LAMP assay. The results showed that this real-time turbidimeter LAMP assay was specific and sensitive enough to amplify DNA from soybean products. The limit of detection (LOD) of lectin and MON89788 was 0.01% (weight of GM soybean/weight of non-GM soybean), roughly equivalent to four copies of soybean genomic DNA in DNA mixture within one hour; and the LOD of nos and GTS40-3-2 was 0.1%, roughly equivalent to forty copies of soybean genomic DNA within 1 hour. The sensitivity reaches to 0.01%, which meets the requirements of the European Union (EU). In practical application, the total detectable ratio in soybean-made products was the same as that of PCR, suggesting that LAMP method was accurate and sensitive.

Keywords:
real-time turbidimeter-LAMP GM soybean rapid detection

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