Electrochemical magnetoassay coupled to PCR as a quantitative approach to detect the soybean transgenic event GTS40-3-2 in foods
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Elsevier
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Simple, cost-effective and reliable tools for the quantification of genetically modified organisms (GMO) in food and feed are highly demanded to enforce labelling legislation in the EU. Herein, we report a novel method for quantitative analysis of genetically modified soybean with the event GTS-40-3-2, also known as Roundup Ready (RR) soybean, using magnetoassays with electrochemical detection, coupled to DNA amplification by end-point polymerase chain reaction (PCR). For the proposed work, two DNA sequences were targeted via hybridisation onto magnetic beads, one specific for the transgenic event and the other for the taxon or species-specific lectin gene. Enzymatic labelling was performed to obtain an electrochemically active product measured by chronoamperometry. By optimising the number of PCR cycles, among other parameters, two magnetoassays coupled to PCR were successfully accomplished and linearity was obtained in the ranges of 53–4425 and 1093–88,496 DNA copies for the event-specific and lectin sequences, respectively. The proposed method provides accurate and precise RR soybean quantitative results, being effectively compared to those obtained by real-time PCR, as the reference method. These findings confirm the suitability of the method as an alternative tool for GMO quantification
Simple, cost-effective and reliable tools for the quantification of genetically modified organisms (GMO) in food and feed are highly demanded to enforce labelling legislation in the EU. Herein, we report a novel method for quantitative analysis of genetically modified soybean with the event GTS-40-3-2, also known as Roundup Ready (RR) soybean, using magnetoassays with electrochemical detection, coupled to DNA amplification by end-point polymerase chain reaction (PCR). For the proposed work, two DNA sequences were targeted via hybridisation onto magnetic beads, one specific for the transgenic event and the other for the taxon or species-specific lectin gene. Enzymatic labelling was performed to obtain an electrochemically active product measured by chronoamperometry. By optimising the number of PCR cycles, among other parameters, two magnetoassays coupled to PCR were successfully accomplished and linearity was obtained in the ranges of 53–4425 and 1093–88,496 DNA copies for the event-specific and lectin sequences, respectively. The proposed method provides accurate and precise RR soybean quantitative results, being effectively compared to those obtained by real-time PCR, as the reference method. These findings confirm the suitability of the method as an alternative tool for GMO quantification
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This work was supported by Fundacão para a Ciência e a Tecnologia (FCT) through grant no. PEst C/EQB/LA0006/2013 financed by POPH-QREN (subsidised by FSE and MCTES), by the bilateral project Acc¸ ões Integradas Luso-Espanholas/2012 between University of Porto (E-38/12) and University of Oviedo (Ministerio de Ciencia e Innovación, PRI-AIBPT-2011-0769), CTQ2012-31157 and by Marie Curie Actions FP7-PEOPLE-2013-IRSES through the project no. 612545 entitled “GMOsensor–Monitoring Genetically Modified Organisms in Food and Feed by Innovative Biosensor Approaches”. C. L. Manzanares-Palenzuela is grateful to Universidad Complutense de Madrid for a pre-doctoral grant (BE43/11). J. Costa and M. F. Barroso are grateful to FCT for post-doctoral grants (SFRH/BPD/102404/2014 and SFRH/BPD/78845/2011, respectively) financed by POPH-QREN (subsidised by FSE and MCTES)
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