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Development and application of loop-mediated isothermal amplification for detection of genetically modified crops.
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2009
Year
EngineeringConventional PcrNucleic Acid Amplification TestPlant PathologyBioanalysisBiochemical EngineeringAnalytical BiotechnologyMolecular DiagnosticsLoop-mediated Isothermal AmplificationMolecular Biological MethodAgricultural BiotechnologyBiomolecular EngineeringMolecular Diagnostic TechniquesLamp MethodBiotechnologyGenetic EngineeringNucleic Acid AmplificationMicrobiologyMedicineGenetically Modified Organism
【Objective】 The objective of the study is to develop a loop-mediated isothermal amplification method for detection of genetically modified crops(GMC). 【Method】 The detection method is based on the loop-mediated isothermal amplification (LAMP) reaction. Cauliflower mosaic virus 35S (CaMV35S) promoter gene, a widespread genetic element, was amplified by a set of four specially primers that recognize six distinct sequences of the target. The amplification can be obtained in 1 h by incubating all of the reagents in a single tube with Bst DNA polymerase at 65℃. 【Result】 The LAMP method can detect a specific promoter containing CaMV35S from GMC and their test results were consistent with the results of conventional PCR methods. LAMP assay results were found to be 10 times more sensitive than the conventional PCR. 【Conclusion】 The LAMP detection method is specific, stable and reliable, and will be an effective tool for rapid detection of GMC.