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Development of PCR screening assays focused on genecoding sequences for GMO detection


Frédéric Debode, Éric Janssen, Gilbert Berben

For the detection of genetically modifed organisms (GMO), screening is the frst step used to determine if a GMO or its derived products are present in food or feed. If the result is positive, additional tests must be done to identify and, where necessary, quantify the genetically modifed (GM) event present. The screening must be as wide as possible to cover the different possible GM events encountered on the market. In this paper, we present new real-time PCR screening methods (DNA-based methods) using hybridization probes (TaqMan probes) focused on gene-coding regions. These were selected on the basis of their occurrence in GM constructs (Block et al., 2013). Detection is also possible by searching for the new proteins using immunoassays (ELISA or immunochromatographic test strips). These techniques are well-suited for the detection of proteins produced by the EPSPS, pat, bar and cry genes (Stave, 2002; Van den Bulcke et al., 2007). However, this approach is only suitable for raw and unprocessed products. DNA-based methods were therefore preferred in this study. PCR assays focused on genes can be combined with those focused on promoters and terminators (Debode et al., 2013) in order to provide screening with wider coverage. The PCR assays developed in this paper were based on the sequences of the bar, pat, EPSPS, gox, gus and hsp70 genes.

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Published
September, 2018

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