Development of a Microfluidic Chip for Detection of Genetically Modified Organisms by Loop-mediated Isothermal Amplification
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Abstract:
Loop-mediated isothermal amplification (LAMP) has been increasingly applied in the genetic detection of transgenic crops and microorganisms in recent years. In order to improve the detection efficiency of related inspection agencies for transgenic products and speed up the clearance of imported and exported agricultural products and foods, LAMP was combined with a microfluidic chip for the first time to simultaneously detect ten genetic recombinant elements, including P-CAMV35s, T-NOS, NPT, BAR, PAT, Cry1Ac, EPSPSI, EPSPSII, FMV35s, and GA-Hra, in samples at 65°C. The limit of detection could reach 0.5% (m/m), and the detection of transgenic ingredients in the sample could be completed with one sample loading. This method had a good specificity, a high sensitivity, and a short detection time, and the amplification signals could be acquired in real time using software. The detection results for actual samples indicated that all the results were consistent with those obtained by the real-time polymerase chain reaction (PCR) standard method. The microfluidic chip developed in this study for LAMP can be used for the inspection of transgenic ingredients in common agricultural products (soybean, corn, rice, cotton, etc.), and provides a rapid, sensitive, and high-throughput technical platform for the qualitative detection of transgenic products.
