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


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Article Info

DATE

2020-01-22

AUTHORS

Tingting Deng, Wensheng Huang, Ranran Xing, Jiukai Zhang, Junan Ren, Yiqiang Ge, Ying Chen

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. More... »

PAGES

631-641

References to SciGraph publications

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  • 2010-01-20. Comparison of nine different real-time PCR chemistries for qualitative and quantitative applications in GMO detection in ANALYTICAL AND BIOANALYTICAL CHEMISTRY
  • 2015-03-31. GMO detection in food and feed through screening by visual loop-mediated isothermal amplification assays in ANALYTICAL AND BIOANALYTICAL CHEMISTRY
  • 2015-03-11. Collaborative trial validation of cry1Ab/Ac and Pubi-cry TaqMan-based real-time PCR assays for detection of DNA derived from genetically modified Bt plant products in ACCREDITATION AND QUALITY ASSURANCE
  • 2014-05-09. Establishment and application of a loop-mediated isothermal amplification (LAMP) system for detection of cry1Ac transgenic sugarcane in SCIENTIFIC REPORTS
  • 2015-08-04. A highly sensitive and specific method for the screening detection of genetically modified organisms based on digital PCR without pretreatment in SCIENTIFIC REPORTS
  • 2014-03-26. Inter-laboratory Testing of GMO Detection by Combinatory SYBR®Green PCR Screening (CoSYPS) in FOOD ANALYTICAL METHODS
  • 2011-02-20. Construction of a reference plasmid molecule containing eight targets for the detection of genetically modified crops in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2009-10-02. Development of a qualitative, multiplex real-time PCR kit for screening of genetically modified organisms (GMOs) in ANALYTICAL AND BIOANALYTICAL CHEMISTRY
  • 2008-04-24. Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products in NATURE PROTOCOLS
  • 2012-03-24. In-depth analysis of the endogenous reference genes used in the quantitative PCR detection systems for rice in EUROPEAN FOOD RESEARCH AND TECHNOLOGY
  • 2018-10-06. Event-Specific Qualitative and Quantitative Detection of Genetically Modified Rice G6H1 in FOOD ANALYTICAL METHODS
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    URI

    http://scigraph.springernature.com/pub.10.1007/s00217-020-03430-1

    DOI

    http://dx.doi.org/10.1007/s00217-020-03430-1

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