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Establishment and application of a loop-mediated isothermal amplification method with double-stranded displacement probes to quantify the genetically modified rice M12 event

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For the purpose of supervision and academic research on GMO, it was necessary to establish a simple and effective on-site qualitative and quantitative method for GMO detection. In this paper,… Click to show full abstract

For the purpose of supervision and academic research on GMO, it was necessary to establish a simple and effective on-site qualitative and quantitative method for GMO detection. In this paper, double-stranded displacement probes were first designed to monitor the quantitative LAMP amplification, and calcein was used effectively in LAMP reaction for qualitative measurement. High specificity of the methods was identified by testing 16 crops. The rice endogenous gene SPS and M12 event-specific gene can both be quantified in a wide dynamic range (200–20,000 copies). The LOD and LOQ of both genes were 20 and 200 copies, respectively. When the methods were used to analyze rice samples, 0.5% (w/w) of the GM rice M12 event could be quantified accurately and 0.1% (w/w) could be detected qualitatively. These results indicated that the developed methods were specific, sensitive and repeatable, and proven to be an efficient alternative to real-time PCR. Moreover, the developed methods were more suitable for the reliable quantification in poor-equipped laboratories or on-site detection.

Keywords: m12 event; displacement probes; rice m12; double stranded; event; stranded displacement

Journal Title: European Food Research and Technology
Year Published: 2020

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