Optimization of loop-mediated isothermal amplification (LAMP) reaction mixture for biosensor applications

Official URL: https://dx.doi.org/10.1016/j.mex.2021.101282

Genetically Modified (GM) foods are becoming the future of agriculture on surviving global natural disasters and climate change by their enhanced production efficiency and improved functional properties. On the other hand, their adverse health and environmental effects, ample evidence on transgene leakage of Genetically Modified Organisms (GMOs) to crops have raised questions on their benefits and risks. Consequently, low-cost, reliable, rapid, and practical detection of GMOs have been important. GMO-detection platforms should be capable of stably storing detection reagents for long-delivery distances with varying ambient temperatures. In this study, we developed an event-specific, closed tube colorimetric GMO detection method based on Loop-Mediated Isothermal Amplification (LAMP) technique which can be integrated into GMO-detection platforms. The entire detection process optimized to 30 min and isothermally at 65 °C. The durability of the LAMP mixture in the test tubes showed that the LAMP reaction mixture, in which Bst polymerase and DNA sample was later included, yielded DNA amplicons for 3 days at room temperature, and for 6 days at 4 °C. • Simple, stable, and cheap storage method of LAMP reaction mixture for GMO-detection technologies. • GMO-detection platforms can stably store detection reagents for long-delivery distances with varying ambient temperatures. • Any DNA sample can be used in the field or resource-limited setting by untrained personnel.