title: Loop-mediated isothermal amplification: rapid visual and real-time methods for detection of genetically modified crops.
abstract: 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.
authors.affiliation: Division of Genomic Resources (National Research Centre on DNA Fingerprinting), National Bureau of Plant Genetic Resources , New Delhi 110 012, India.
authors.forename: Gurinder Jit
authors.initials: GJ
authors.lastname: Randhawa
authors.forename: Monika
authors.initials: M
authors.lastname: Singh
authors.forename: Dany
authors.initials: D
authors.lastname: Morisset
authors.forename: Payal
authors.initials: P
authors.lastname: Sood
authors.forename: Jana
authors.initials: J
authors.lastname: Zel
mesh_terms: D017796:Caulimovirus / Q000235:genetics; D018556:Crops, Agricultural; D017931:DNA Primers / Q000235:genetics; D005819:Genetic Markers; D005966:Glucuronidase / Q000235:genetics; D003368:Gossypium / Q000235:genetics; D019868:Kanamycin Kinase / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009713:Nucleotidyltransferases / Q000235:genetics; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D005819:Genetic Markers; D019868:Kanamycin Kinase; D009713:Nucleotidyltransferases; C022678:streptomycin 3''-adenylyltransferase; D005966:Glucuronidase