Validation of the Method for Event Specific Mon810 and NK603 and Endogenous Gene Identification by using Multiplex Real-time PCR Technique
DOI:
https://doi.org/10.14456/thaidoa-agres.2019.19Keywords:
genetically modified maize; multiplex real-time PCR technique; Mon810; NK603; validationAbstract
Genetically modified maize has been widely accepted for commercialization. The most popular approved events are lepidopteran insect resistance (Mon810) and glyphosate herbicide tolerance (NK603). However, importation conditions of GM products in some countries requires identification of the GM event and biosafety assessment. At present, the international laboratory analyzes GM event by using simplex real-time PCR technique which is lengthy, contains many steps and is prone to contamination. The multiplex real-time PCR had been developed for detecting multiple modified genes at the same time. We validated this method for GM maize: Mon810 and NK603 events detection. The results revealed that Mon810, NK603 and HMG primers and probes were absolutely specific to Mon810 and NK603 maize. The parameters of this multiplex real-time PCR method were within the acceptable parameter standard including 97 to 112 percentage of PCR efficiency, -3.38 to -3.07 of slope, 0.99 of linearity (R2), limit of detection in 0.1 percentage and no false positive or negative results. In summary, multiplex real-time PCR in this study can be used for the events screening method of GM maize and maize products
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