Loop-Mediated Isothermal Amplification: Rapid Visual and Real-Time Methods for Detection of Genetically Modified Crops
| Autor: | Gurinder Jit Randhawa, Jana Zel, Dany Morisset, Monika Singh, Payal Sood |
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| Rok vydání: | 2013 |
| Předmět: |
Crops
Agricultural Genetic Markers Detection of genetically modified organisms Gossypium Kanamycin Kinase biology Kanamycin kinase Loop-mediated isothermal amplification General Chemistry Nucleic acid amplification technique Genetically modified crops Plants Genetically Modified biology.organism_classification Nucleotidyltransferases Sensitivity and Specificity DNA extraction Molecular biology Genetically modified organism Caulimovirus Cauliflower mosaic virus Promoter Regions Genetic General Agricultural and Biological Sciences Nucleic Acid Amplification Techniques DNA Primers Glucuronidase |
| Zdroj: | Journal of Agricultural and Food Chemistry. 61:11338-11346 |
| ISSN: | 1520-5118 0021-8561 |
| DOI: | 10.1021/jf4030085 |
| Popis: | A rapid, reliable, and sensitive loop-mediated isothermal amplification (LAMP) system was developed for screening of genetically modified organisms (GMOs). The optimized LAMP assays using designed primers target commonly employed promoters, i.e., Cauliflower Mosaic Virus 35S (P-35S) and Figwort Mosaic Virus promoter (P-FMV), and marker genes, i.e., aminoglycoside 3'-adenyltransferase (aadA), neomycin phosphotransferase II (nptII), and β-glucuronidase (uidA). The specificity and performance of the end-point and real-time LAMP assays were confirmed using eight genetically modified (GM) cotton events on four detection systems, employing two chemistries. LAMP assays on the isothermal real-time system were found to be most sensitive, detecting up to four target copies, within 35 min. The LAMP assays herein presented using alternate detection systems can be effectively utilized for rapid and cost-effective screening of the GM status of a sample, irrespective of the crop species or GM trait. These assays coupled with a fast and simple DNA extraction method may further facilitate on-site GMO screening. |
| Databáze: | OpenAIRE |
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