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Loop-mediated isothermal amplification: rapid visual and real-time methods for detection of genetically modified
Gurinder Jit Randhawa1, Monika Singh, Dany Morisset
1Division of Genomic Resources (National Research Centre on DNA Fingerprinting), National Bureau of Plant Genetic Resources , New Delhi 110 012, India.
Journal of Agricultural and Food Chemistry
|November 6, 2013
Summary
A new loop-mediated isothermal amplification (LAMP) method rapidly screens genetically modified organisms (GMOs). This cost-effective technique offers sensitive detection for on-site GMO testing.
Area of Science:
- Biotechnology
- Molecular Biology
- Food Science
Background:
- Genetically modified organisms (GMOs) require reliable detection methods.
- Existing screening methods can be time-consuming and expensive.
- Isothermal amplification offers potential for rapid, field-deployable diagnostics.
Purpose of the Study:
- To develop and validate a rapid, sensitive, and reliable loop-mediated isothermal amplification (LAMP) system for GMO screening.
- To target commonly used genetic elements in GMOs, including promoters and marker genes.
- To assess the performance of LAMP assays across different detection systems and chemistries.
Main Methods:
- Designed and optimized LAMP primers targeting Cauliflower Mosaic Virus 35S (P-35S) and Figwort Mosaic Virus promoter (P-FMV).
- Targeted marker genes: aminoglycoside 3'-adenyltransferase (aadA), neomycin phosphotransferase II (nptII), and β-glucuronidase (uidA).
- Evaluated end-point and real-time LAMP assays using eight genetically modified (GM) cotton events on four detection systems with two chemistries.
Main Results:
- Real-time LAMP assays demonstrated high sensitivity, detecting down to four target copies within 35 minutes.
- Assays confirmed specificity and performance across various GM cotton events and detection platforms.
- The developed LAMP system proved effective for detecting common GMO promoter and marker gene sequences.
Conclusions:
- The developed LAMP assays provide a rapid, cost-effective, and sensitive method for GMO screening.
- These assays are versatile and applicable irrespective of crop species or specific GM trait.
- Integration with simple DNA extraction methods can facilitate on-site GMO detection.
