Rapid Screening Detection of Genetically Modified Crops by Loop-Mediated Isothermal Amplification with a Lateral Flow Dipstick
| Autor: | Kosuke Nakamura, Yukari Kagiya, Reona Takabatake, Keisuke Soga, Junichi Mano, Kazumi Kitta, Yasutaka Minegishi, Kazunari Kondo, Satoshi Futo |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Crops Agricultural Loop-mediated isothermal amplification Genetically modified crops 01 natural sciences Zea mays 03 medical and health sciences Multiplex Plant Proteins Detection limit biology Chemistry 010401 analytical chemistry General Chemistry Dipstick biology.organism_classification Plants Genetically Modified Molecular biology 0104 chemical sciences Genetically modified organism 030104 developmental biology Cry1Ac Cauliflower mosaic virus Soybeans General Agricultural and Biological Sciences Nucleic Acid Amplification Techniques |
| Zdroj: | SC30201812190005 NARO成果DBa 表示-非営利-改変禁止(CC-BY-NC-ND) |
| ISSN: | 1520-5118 |
| Popis: | We developed a novel loop-mediated isothermal amplification (LAMP)-based detection method using lateral flow dipstick chromatography for genetically modified (GM) soybean and maize events. The single-stranded tag hybridization (STH) for the chromatography printed-array strip (C-PAS) system was used for detections targeting the cauliflower mosaic virus 35S promoter, mannose-6-phosphate isomerase gene, Pisum sativum ribulose 1, 5-bisphosphate carboxylase terminator, a common sequence between the Cry1Ab and Cry1Ac genes, and a GA21-specific sequence. The STH C-PAS system was applicable for multiplex analyses to perform simultaneous detections. The limit of detection was 0.5% or less for each target. By using the developed method, the LAMP amplification was visually detected. Moreover, the detection could be carried out without any expensive instruments, even for the DNA amplification steps, by virtue of the isothermal reaction. We demonstrated that the rapid and useful method developed here would be applicable for screening GM crops. |
| Databáze: | OpenAIRE |
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