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Rapid Screening for Detecting Genetically Modified Wheat by Loop-Mediated Isothermal Amplification.

Shinya Kimata1, Takumi Daimon1, Miwa Takahashi1

  • 1QE (Quality Exam.) Center, Research and Development, Quality Assurance Division, Nisshin Seifun Group Inc., 5-3-1 Tsurugaoka, Fujimino, Saitama 356-8511, Japan.

Biological & Pharmaceutical Bulletin
|March 26, 2026
PubMed
Summary

A new loop-mediated isothermal amplification (LAMP) method efficiently screens genetically modified (GM) wheat for glyphosate tolerance. This rapid detection prevents unauthorized GM crop contamination in food and the environment.

Keywords:
genetically modifiedlateral flow DNA chromatographyloop-mediated isothermal amplificationrapid detectionscreeningwheat

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Area of Science:

  • Agricultural Biotechnology
  • Molecular Biology
  • Food Safety

Background:

  • Genetically modified (GM) crops require reliable detection methods for regulatory compliance and safety.
  • Glyphosate tolerance is a common trait in GM crops, necessitating specific screening techniques.
  • Existing methods for GM detection can be time-consuming or require specialized equipment.

Purpose of the Study:

  • To develop a rapid and sensitive loop-mediated isothermal amplification (LAMP) method for screening glyphosate-tolerant GM wheat.
  • To establish a detection system that can identify specific genetic elements associated with glyphosate tolerance.
  • To differentiate between common and durum wheat varieties using endogenous reference gene detection.

Main Methods:

  • Development of LAMP assays targeting the cauliflower mosaic virus 35S promoter (P35S) and the EPSPS gene from Agrobacterium tumefaciens CP4.
  • Utilized a real-time fluorometer (Genie II) for LAMP product detection.
  • Designed primer sets for wheat endogenous reference genes (proline-rich protein [PRP] and Waxy-D1).
  • Employed single-stranded tag hybridization (STH) on a chromatography printed-array strip (C-PAS) system for enhanced detection efficiency.

Main Results:

  • The LAMP method achieved a limit of detection (LOD) of ≤0.05% (14 copies of target DNA).
  • Primer sets for PRP and Waxy-D1 enabled differentiation between common and durum wheat.
  • The STH C-PAS system successfully detected LAMP products at the LOD.
  • Duplex detection of P35S/PRP and EPSPS/PRP was achieved, demonstrating multiplexing capability.

Conclusions:

  • A simple, rapid, and sensitive LAMP-based method was successfully developed for screening glyphosate-tolerant GM wheat.
  • The STH C-PAS system enhances the efficiency and reliability of GM wheat detection.
  • This method offers a valuable tool for preventing the environmental and food supply contamination by unauthorized GM crops.