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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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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.

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Abbreviations

LAMP:

Loop-mediated isothermal amplification

GM:

Genetically modified

PCR:

Polymerase chain reaction

LOD:

Limit of detection

LOQ:

Limit of quantification

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Acknowledgements

This work was supported by National GMO Cultivation Major Project of New Varieties (Grant no. 2018ZX08012001).

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Correspondence to Yiqiang Ge or Ying Chen.

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The authors declare no conflict of interest.

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Deng, T., Huang, W., Xing, R. et al. Establishment and application of a loop-mediated isothermal amplification method with double-stranded displacement probes to quantify the genetically modified rice M12 event. Eur Food Res Technol 246, 631–641 (2020). https://doi.org/10.1007/s00217-020-03430-1

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Keywords

  • Loop-mediated isothermal amplification (LAMP)
  • Rice
  • Genetically modified rice M12 event
  • Quantification