Kaymaz Sümeyra Vural, Elitas Meltem
Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul 34956, Turkey.
MethodsX. 2021 Feb 19;8:101282. doi: 10.1016/j.mex.2021.101282. eCollection 2021.
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 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.
转基因(GM)食品因其提高的生产效率和改善的功能特性,正成为农业应对全球自然灾害和气候变化的未来发展方向。另一方面,它们对健康和环境的不利影响,以及大量关于转基因生物(GMO)向作物发生转基因泄漏的证据,引发了人们对其益处和风险的质疑。因此,低成本、可靠、快速且实用的转基因生物检测至关重要。转基因生物检测平台应能够在不同环境温度下,将检测试剂稳定储存以用于长距离运输。在本研究中,我们基于环介导等温扩增(LAMP)技术开发了一种针对特定事件的、闭管比色法转基因生物检测方法,该方法可集成到转基因生物检测平台中。整个检测过程在65℃等温条件下优化至30分钟。试管中LAMP混合物的耐用性表明,在其中随后加入聚合酶和DNA样品的LAMP反应混合物,在室温下可产生DNA扩增子3天,在4℃下可产生6天。
•用于转基因生物检测技术的LAMP反应混合物的简单、稳定且廉价的储存方法。
•转基因生物检测平台能够在不同环境温度下,将检测试剂稳定储存以用于长距离运输。
•未经培训的人员可在现场或资源有限的环境中使用任何DNA样品。