Assessment of the Real-Time PCR Method Claiming to be Specific for Detection and Quantification of the First Commercialised Genome-Edited Plant

• DOI: 10.1007/s12161-022-02237-y
• Publication Date: 2022-04-04T00:02:17Z
• Authors: Christopher Weidner; Sophia Edelmann; Dominik Moor; Kathrin Lieske; Christian Savini; Sara Jacchia; Maria Grazia Sacco; Marco Mazzara; Jörn Lämke; Kolja Neil Eckermann; Hendrik Emons; Joachim Mankertz; Lutz Grohmann
• Keywords: genomic DNA

Abstract A real-time PCR method was recently published with a claim to be specific for the detection and identification of some genome-edited oilseed rape (OSR) lines commercialised in North America. The designed detect single base mutation AHAS1C gene, which confers herbicide tolerance. authors that is event-specific OSR line 5715 fulfils all requirements GMO analytical methods according EU regulations. We have thoroughly assessed relation minimum performance (MPR) established by European Network Laboratories (ENGL). found sufficiently sensitive robust when tested pure genomic DNA 40 K. However, our results show not detects also carrying same point caused somaclonal variation. Moreover, impaired robustness observed using non-modified at amount specified original protocol. Significant non-specific amplifications products as non-target template from numerous varieties well wild radish were three ISO/IEC 17025 accredited reference laboratories tests different master mixes cycler models. assessment shows does meet MPR qualitative therefore fit-for-purpose official controls genetically modified EU. Suggestions are provided conditions under organisms should validated.