Event-Specific Plasmid Standards and Real-Time PCR Methods for Transgenic Bt11, Bt176, and GA21 Maize and Transgenic GT73 Canola

Taverniers, Isabel; Windels, Pieter; Vaïtilingom, Marc; Milcamps, Anne; Bockstaele, Erik Van; Eede, Guy Van den; Loose, Marc De

doi:10.1021/jf0483467

Cited by 102 publications

(67 citation statements)

References 32 publications

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“…The obtained detection limits of our developed MLPA assay are in the range of a rapeseed qPCR [24] [25]. One should keep in mind that we used genomic DNA as starting material, whereas plasmids are often used as calibrators in qPCR [26]. By doing so, the possibility of contamination is reduced and potential matrix effects are also taken into account.…”

Section: Discussionmentioning

confidence: 97%

Development of a Multiplex Ligation Dependent Probe Amplification (MLPA) Module for Simultaneous Detection of Five Genetically Modified Rapeseed Events

Guertler¹,

Goerlich²,

Busch³

2014

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For the official surveillance of genetically modified crops, efficient and simple detection methods need to be on hand to implement the strict requirements regarding approval, labelling and traceability determined by the European Union. Therefore, a multiplex ligation dependent probe amplification (MLPA) module was developed for simultaneous detection of five genetically modified rapeseed events (MS8, RF3, GT73, Falcon GS40/90 and T45). Probes were designed and concentrations were adapted in order to obtain high sensitivity and specificity. This MLPA module was validated using certified reference materials and its applicability was tested analyzing routine honey, mustard and rapeseed samples. The limit of detection was determined by analyzing a dilution series (n = 16 for each concentration) of respective transgenic DNA. After optimization, the MLPA revealed limits of detection between 10 to 50 copies of the transgene DNA/assay. The method proved to be sensitive and highly reproducible. When analyzing routine samples, results obtained applying the MLPA module were similar compared to real-time PCR.

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Section: Discussionmentioning

confidence: 97%

Development of a Multiplex Ligation Dependent Probe Amplification (MLPA) Module for Simultaneous Detection of Five Genetically Modified Rapeseed Events

Guertler¹,

Goerlich²,

Busch³

2014

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“…be confirmed in detail using some reference methods (PCR 40,41 or protein immunoassays) and much larger number of samples in order to obtain statistically significant data, and also some information about possible quantification aspects of such measurements. So far, the main obstacle in such studies is difficulty in obtaining model samples of different GMO variants, even for the most commonly cultivated maize or soybean.…”

Section: Modification Of Capillary Wall With Polybrenementioning

confidence: 98%

Modification of Resolution in Capillary Electrophoresis for Protein Profiling in Identification of Genetic Modification in Foods

et al. 2011

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Abstract. The capillary electrophoresis with UV detection was employed for protein profiling in extracts from maize and soybeans. Modifications of back-ground electrolyte and coating the capillary wall with polybrene was employed in order to decrease the protein adsorption on the capillary walls. The obtained protein profiles were compared for transgenic and non-transgenic variants, showing in some cases significant changes that might be employed for identification of genetic modifications of plants or foods of plant origin. (doi: 10.5562/cca1777)

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“…-Event-specific quantitative -ImmunoStrip and PCR (Taverniers et al 2005) ELISA (Agdia, leaves only).…”

Section: Cultivars Commercially (Resistance To Europeanmentioning

confidence: 99%

Adventitious presence of GMOs: Scientific overview for Canadian grains

Demeke¹,

Perry²,

Scowcroft³

2006

Can. J. Plant Sci.

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. 2006. Adventitious presence of GMOs: Scientific overview for Canadian grains. Can. J. Plant Sci. 86: 1-23. The global expansion in the development and cultivation of genetically modified (GM) crops has increased international concern about adventitious presence of GM materials in non-GM seeds and grains. GM events in canola, corn, soybean, cotton, flax, papaya, potato, squash, sugar beet, and tomato have received regulatory approval in Canada. However, GM cultivars are only in commercial production for canola, corn and soybean. More than 30 GM events have been approved in these three crops. Cases of unapproved adventitious presence of GM materials that have impacted grain trading and handling in Canada and other countries include StarLink™ corn, GT200 canola, GM canola in mustard and recently Bt10 corn. Some countries have established tolerance and traceability requirements for adventitious presence of GMOs, while others are in the process of developing or adopting legislation. The threshold for labeling of adventitious presence of approved GM material in non-GM grain varies from 0.9% (e.g., EU) to 5% (e.g., Japan). Progress has been made in the development of DNA-and proteinbased GMO detection methods. However, only a limited number of these detection methods have been internationally validated. The challenges for detection methods include sampling, a lack of certified reference material, a lack of DNA sequence information for the design of event-specific primers, and the sheer number of individual events that may be present and tested for. Current efforts by ISO and CEN will be valuable for establishing harmonized and standardized GMO detection methods.Key words: List of GM events, cases of AP, tolerance, traceability, detection methods, challenges Demeke, T., Perry, D. J. et Scowcroft, W. R. 2006. Présence de matériel génétiquement modifié adventif : survol de la situation des grains au Canada. Can. J. Plant Sci. 86: 1-23. Le développement et la culture de matériel génétiquement modifié (GM) connaissent une expansion mondiale si bien que, partout sur terre, on s'inquiète de la présence de matériel GM adventif dans les semences et les grains non GM. La modification génétique du canola, du maïs, du soja, du coton, du lin, de la papaye, de la pomme de terre, de la courge, de la betterave sucrière et de la tomate a été autorisée par les organismes de réglementation au Canada. Les cultivars GM cultivés commercialement se limitent néanmoins au canola, au maïs et au soja. Plus de 30 modifications génétiques ont été approuvées pour ces trois cultures. Parmi les cas de matériel GM adventif non autorisé qui ont posé des problèmes au niveau des échanges commerciaux et de la manutention du grain au Canada et ailleurs, on retrouve le maïs StarLink MC , le canola GT200, le canola GM dans la moutarde et, plus récemment, le maïs Bt10. Certains pays tolèrent ou exigent qu'on puisse retracer le matériel GM adventif, mais d'autres ont entrepris l'élaboration ou l'adoption de lois en la matière. Le seuil de tolérance concernan...

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Event-Specific Plasmid Standards and Real-Time PCR Methods for Transgenic Bt11, Bt176, and GA21 Maize and Transgenic GT73 Canola

Cited by 102 publications

References 32 publications

Development of a Multiplex Ligation Dependent Probe Amplification (MLPA) Module for Simultaneous Detection of Five Genetically Modified Rapeseed Events

Development of a Multiplex Ligation Dependent Probe Amplification (MLPA) Module for Simultaneous Detection of Five Genetically Modified Rapeseed Events

Modification of Resolution in Capillary Electrophoresis for Protein Profiling in Identification of Genetic Modification in Foods

Adventitious presence of GMOs: Scientific overview for Canadian grains

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