Abstract
Combinatory SYBR®Green real-time PCR Screening (CoSYPS) is an efficient, sensitive approach for detecting complex targets such as genetically modified organisms (GMOs) in food and feed products. GMO analysis for legal purposes has become increasingly complex and costly due to the diversity in recombinant targets present in the different GMOs. For this reason, screening for the presence of GMOs is in general the first step in the detection of GM material in a product. CoSYPS allows detecting the large majority of globally commercial GMOs using SYBR®Green real-time PCR methods for six GM targets (P35S, Tnos, CryIAb, CP4-EPSPS, PAT and BAR) combined with species-specific PCR methods (e.g., maize, soy, rapeseed). Here, the results of an inter-laboratory trial on seven samples with different GMO mixtures at different levels are presented. In total, 13 laboratories participated in the trial and the currently most frequently used PCR analysis platforms are represented. The inter-laboratory study clearly demonstrates that PCR methods used in CoSYPS form a very robust GMO screening system. Sensitivity, specificity, positive and negative predictive values are for all PCR methods higher than 95 % for all samples. Together, these results show that the SYBR®Green real-time PCR methods used in CoSYPS are effectively applicable to different PCR platforms and amendable to configuration into a sensitive high-throughput GMO screening and decision support tool.
Highlights
Modified (GM) food and feed products have become a reality today (James 2012)
The aim of this study was dual: first, to demonstrate a “proof of concept” of Combinatory SYBR®Green real-time PCR Screening (CoSYPS) to detect various levels of genetically modified organisms (GMOs) in different backgrounds and the evaluation of pre-fixed “cut-off values” and the CoSYPS decision support system (DSS); and second, to demonstrate the general applicability of CoSYPS to estimate the impact of introduction of SYBR®Green quantitative PCR (qPCR) analysis and the potential adaptive actions to take in the context of implementation of the CoSYPS methodology in a new laboratory
All laboratories were satisfied with the included protocols, 31 % of the participants considered the transfer of the analytical results into the CoSYPS Decision Support System as “tedious”
Summary
Modified (GM) food and feed products have become a reality today (James 2012) Since their initial commercialization in 1994, about 150 genetically modified plants have received approval for use as food or feed (http://ceragmc.org). Considering their large diversity (different GM elements in various species), the application of a generic screening for the presence of GM material is most often the first step in GMO analysis (Holst-Jensen et al 2012). Such qualitative screening methods provide a presence/absence response (taxon-specific sequence or a GM element) and help reducing the number of subsequent identification analyses
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