SYBR®Green qPCR methods for the detection of the Roundup Ready® Cp4-EPSPS, LibertyLink® “PAT” and “BAR” and the Bacillus thuringiensis CryIAb/Ac traits as present in genetically modified organisms (GMO) were developed. The specificity, sensitivity, dynamic range and PCR method efficiency was determined for each of them. All methods comply with the performance criteria set by the European Network of GMO Laboratories (ENGL), ISO norms and Codex guidelines. The developed methods are specific and generate amplicons of 108, 73, 109 and 69 bp respectively for “Cp4-EPSPS”, “CryIAb/Ac”, “PAT” and “BAR” targets. Single-target Sybricon plasmids comprising the corresponding PCR amplicons have been constructed and used as controls in PCR analysis. These four SYBR®Green qPCR methods for GMO analysis complete the set of qPCR methods applied in the so-called combinatory SYBR®Green qPCR screening (abbreviated as "CoSYPS''; Van den Bulcke et al, 2010, Anal Bioanal Chem, 396(6):2113-23). Due to their trait-specific nature, these methods allow discriminating better between the different GMO. Compared to GMO screening applying only common targets such as the 35S promoter or the NOS terminator, screening with trait-specific methods reduces the number of possible GMO candidates present in a sample, reducing the cost of the analysis. The application of these methods in CoSYPS analysis is demonstrated using two GEMMA proficiency test samples and reference material from the genetically modified rapeseed event GT73. This set of SYBR®Green qPCR trait-specific methods represents a very interesting novel set of GMO analysis methods allowing cost-effective identification of GM materials in products.

BARBAU-PIEDNOIR Elodie;  LIEVENS Antoon;  VANDERMASSEN Els;  MBONGOLLO MBELLA Etondoh-Guillaume;  LEUNDA-CASI Amaya;  ROOSENS Nancy;  SNEYERS Myriam;  VAN DEN BULCKE Marc; 

2012-12-14

SPRINGER

JRC76623

1438-2377,   

https://publications.jrc.ec.europa.eu/repository/handle/JRC76623,   

10.1007/s00217-011-1605-7,