Does DNA extraction affect the specificity of a PCR Method claiming the specific detectability of a genome-edited plant?

Under current EU legislation, genetically modified organisms (GMOs) and derived food and feed products must be authorised as GM food, feed or seed and appropriate detection methods must be made available for use in official controls. A Real-Time PCR method has recently been published by Chhalliyil et al. claiming to be specific for the detection and identification of genome-edited oilseed rape (OSR) lines commercialised in North America. We have independently assessed this method in three reference laboratories for sensitivity, specificity and robustness. We found that the method does not meet all the minimum performance requirements (MPR) for GMO testing in the EU, which contradicts the results of the method developer. Here we show, in addition to the previously published method assessment, that a modified DNA extraction is not the reason for the contradictory results and does not affect the specificity of the method. We also discuss the procedures recently proposed by the method developers for interpreting PCR results with high Cq values.

SOPHIA Edelmann;  SAVINI Cristian;  MOOR Dominik;  LÄMKE Jörn;  LIESKE Kathrin;  MAZZARA Marco;  EMONS Hendrik;  MANKERTZ Joachim;  WEIDNER Christopher; 

2024-11-26

TAYLOR & FRANCIS AS

JRC137284

2164-5698 (online),   

https://www.tandfonline.com/doi/citedby/10.1080/21645698.2024.2423441?scroll=top&needAccess=true,    https://doi.org/10.1080/21645698.2024.2423441,    https://publications.jrc.ec.europa.eu/repository/handle/JRC137284,   

10.1080/21645698.2024.2423441 (online),