Rapid Screening Detection of Genetically Modified Crops by Loop-Mediated Isothermal Amplification with a Lateral Flow Dipstick

J Agric Food Chem. 2018 Jul 25;66(29):7839-7845. doi: 10.1021/acs.jafc.8b01765. Epub 2018 Jul 11.

Abstract

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.

Keywords: dipstick DNA chromatography; genetically modified (GM); loop-mediated isothermal amplification (LAMP); multiplex analysis; rapid.

Publication types

  • Evaluation Study

MeSH terms

  • Crops, Agricultural / chemistry
  • Crops, Agricultural / genetics
  • Glycine max / chemistry
  • Glycine max / genetics*
  • Nucleic Acid Amplification Techniques / methods*
  • Plant Proteins / genetics
  • Plants, Genetically Modified / chemistry
  • Plants, Genetically Modified / genetics*
  • Zea mays / chemistry
  • Zea mays / genetics*
  • Zea mays / metabolism

Substances

  • Plant Proteins