Molecular Characterization and Event-Specific Real-Time PCR Detection of Two Dissimilar Groups of Genetically Modified Petunia (Petunia x hybrida) Sold on the Market
Voorhuijzen, Marleen M. ; Prins, Theo W. ; Belter, Anke ; Bendiek, Joachim ; Brünen-Nieweler, Claudia ; Dijk, Jeroen P. van; Goerlich, Ottmar ; Kok, Esther J. ; Pickel, Benjamin ; Scholtens, Ingrid M.J. ; Stolz, Andrea ; Grohmann, Lutz - \ 2020
Frontiers in Plant Science 11 (2020). - ISSN 1664-462X
amplification of linearly-enriched fragments - event-specific detection - genetically modified (GM) - MinION sequencing - petunia - real-time PCR - unauthorized GMO
Petunia plants with unusual orange flowers were noticed on the European market and confirmed to be genetically modified (GM) by the Finnish authorities in spring 2017. Later in 2017, inspections and controls performed by several official laboratories of national competent authorities in the European Union detected several GM petunia varieties with orange flowers, but also another group of unusually colored flowers. In the latter group, a so far undetected gene coding for a flavonoid 3’5’ hydroxylase (F3’5’H) responsible for the purple color was identified by German and Dutch authorities, suggesting that the petunias found on the markets contain different genetic constructs. Here, a strategy is described for the identification of GM petunia varieties. It is based on an initial GMO screening for known elements using (real-time) PCR and subsequent identification of the insertion sites by a gene walking-like approach called ALF (amplification of linearly-enriched fragments) in combination with Sanger and MinION sequencing. The results indicate that the positively identified GM petunias can be traced back to two dissimilar GM events used for breeding of the different varieties. The test results also confirm that the transgenic petunia event RL01-17 used in the first German field trial in 1991 is not the origin of the GM petunias sold on the market. On basis of the obtained sequence data, event-specific real-time PCR confirmatory methods were developed and validated. These methods are applicable for the rapid detection and identification of GM petunias in routine analysis. In addition, a decision support system was developed for revealing the most likely origin of the GM petunia.
Collateral damage? Small-scale fisheries in the global fight against IUU fishing
Song, Andrew M. ; Scholtens, Joeri ; Barclay, Kate ; Bush, Simon R. ; Fabinyi, Michael ; Adhuri, Dedi S. ; Haughton, Milton - \ 2020
Fish and Fisheries 21 (2020)4. - ISSN 1467-2960 - p. 831 - 843.
catch certification - developing countries - fisheries governance - maritime security - organized crime - seafood trade
Concern over illegal, unreported and unregulated (IUU) fishing has led to a number of policy, trade and surveillance measures. While much attention has been given to the impact of IUU regulation on industrial fleets, recognition of the distinct impacts on small-scale fisheries is conspicuously lacking from the policy and research debate. In this paper, we outline three ways in which the application of IUU discourse and regulation undermines small-scale fisheries. First, the mainstream construction of “illegal,” “unreported” and “unregulated” fishing, and also the categorical use of “IUU” in an all-inclusive sense, disregards the diversity, legitimacy and sustainability of small-scale fisheries practices and their governing systems. Second, we explore how the recent trade-related measures to counter IUU fishing mask and reinforce existing inequalities between different sectors and countries, creating an unfair burden on small-scale fisheries and countries who depend on them. Third, as IUU fishing is increasingly approached as “organized crime,” there is a risk of inappropriately targeting small-scale fisheries, at times violently. Reflecting on these three trends, we propose three strategies by which a more sensitive and ultimately more equitable incorporation of small-scale fisheries can be supported in the global fight against IUU fishing.
National Reference Laboratories WFSR : Annual report 2018
Noordam, M.Y. ; Silletti, E. ; Alewijn, A. ; Scholtens, I.M.J. ; Jong, J. de; Raamsdonk, L. aan; Lasaroms, J.J.P. ; Gerssen, A. ; Lee, M.K. van der; Mol, J.G.J. ; Leeuwen, S.P.J. van - \ 2019
Wageningen : Wageningen University & Research (WFSR-report 2019.011) - 51
Bridging legal requirements and analytical methods : a review of monitoring opportunities of animal proteins in feed
Raamsdonk, Leo W.D. van; Prins, Theo W. ; Meijer, Nathan ; Scholtens, Ingrid M.J. ; Bremer, Monique G.E.G. ; Jong, Jacob de - \ 2019
Food Additives & Contaminants. Pt. A, Chemistry, Analysis, Control, Exposure & Risk Assessment 36 (2019)1. - ISSN 1944-0049 - p. 46 - 73.
biological relationships - European legislation - monitoring methods - prion diseases
Availability and safety of food ranks among the basic requirements for human beings. The importance of the food producing sector, inclusive of feed manufacturing, demands a high level of regulation and control. This paper will present and discuss the relationships in the triangle of legislation, the background of hazards with a biological nature, and opportunities for monitoring methods, most notable for prion-based diseases as primary issue. The European Union legislation for prevention of prion-based diseases since 2000 is presented and discussed. The definitions and circumscriptions of groups of species will be analysed in the view biological classification and evolutionary relationships. The state of the art of monitoring methods is presented and discussed. Methods based on visual markers (microscopy), DNA-based methods (PCR), protein-based methods (ELISA, mass spectroscopy, proteomics), near infrared oriented methods and combinations thereof are being evaluated. It is argued that the use in legislation of non-homogeneous groups of species in a biological sense will hamper the optimal design of monitoring methods. Proper definitions are considered to act as bridges between legal demands and suitable analytical methods for effective monitoring. Definitions including specified groups of species instead of single species are more effective for monitoring in a range of cases. Besides the desire of precise circumscription of animal groups targeted by legislation, processed products need well defined definitions as well. Most notable examples are blood versus blood products, and hydrolysis of several types of material. The WISE principle for harmonising the design of legislation and of analytical methods is discussed. This principle includes the elements Witful (reasonable legal principles), Indicative (clear limits between prohibition and authorisation), Societal demands (public health, environment, economy), and Enforceable (presence of suited monitoring methods) in order to promote a balanced effort for reaching the desired level of safety in the food production chain.
Applicability of the poultry qPCR method to detect DNA of poultry processed animal protein materials
Scholtens, Ingrid M.J. ; Prins, Theo W. ; Margry, Rob J.C.F. ; Dahlmans, Harald ; Raamsdonk, Leo W.D. van - \ 2019
Food Control 96 (2019). - ISSN 0956-7135 - p. 53 - 58.
Bovine spongiform encephalopathy (BSE) - Chicken - Duck - Feed - Geese - Poultry - Processed animal proteins (PAPs) - Sensitivity - Species-to-species ban - Transmissible spongiform encephalopathy (TSE) - Turkey
After the Bovine Spongiform Encephalitis (BSE) crisis most processed animal proteins (PAPs) were banned from use in animal feed. For the foreseen reintroduction of pork PAPs in poultry feed, and poultry PAPs in pork feed and to comply with the species-to-species ban that prohibits cannibalism, a sensitive and specific TaqMan PCR detection method for poultry DNA has been designed and published. This poultry method is able to detect DNA of chicken, turkey, duck and geese in one PCR reaction. PAPs however, are a difficult and variable matrix. Therefore, the usability of the poultry method was investigated on a range of different poultry PAPs. It was shown that the poultry detection method is capable of detecting poultry DNA in eight out of nine different poultry PAPs mixed at a 0.1% level in chicken feed. The method can also detect at least 0.1% poultry PAPs mixed in pork PAPs. These results show that the poultry method fulfils the 0.1% detection limit requirement in the EU legislation.
Perceptions of Dutch health care professionals on weight gain during chemotherapy in women with breast cancer
Kruif, J.Th.C.M. de; Scholtens, M.B. ; Rijt, J. van der; Boer, M.R. de; Berg, M.M.G.A. van den; Vries, Y.C. de; Winkels, R.M. ; Visser, M. ; Kampman, E. ; Westerman, M.J. - \ 2019
Supportive Care in Cancer 27 (2019)2. - ISSN 0941-4355 - p. 601 - 607.
Breast cancer - Dietary intake - Health care professionals - Health risks - Physical activity - Weight gain
Purpose: Dutch Health care professionals (HCPs) provide little information concerning health risks associated with weight gain during chemotherapy for breast cancer. Women with breast cancer have specified the need for more information on nutrition and physical activity to deal with weight gain. The aims of this study were to assess the perceptions of Dutch HCPs on weight gain during chemotherapy and in addition evaluate whether and what kind of information on dietary intake and physical activity HCPs provide to prevent/treat weight gain during (neo)adjuvant chemotherapy. Methods: A qualitative study was conducted using semi-structured interviews with 34 HCPs involved in breast cancer care: general practitioners, oncologists, specialized nurses, and dieticians. Results: To date, little information about nutrition, physical activity, and weight gain is given during chemotherapy because it is not part of most HCPs’ training, it is not included in the guidelines and it is not the best time to bring up information in the opinion of HCPs. Weight gain was perceived as just a matter of a few kilos and not an important health issue during treatment. All HCPs felt it is better that women themselves addressed their weight gain after chemotherapy. Conclusion: More knowledge about health risks associated with chemotherapy-induced weight gain and how to combat these issues needs to be made readily available to the HCPs and should become part of their training. Existing patient guidelines should include information on how to prevent and/or reduce weight gain through self-management of nutrition intake and physical activity during and post chemotherapy.
National Reference Laboratories RIKILT : annual report 2017
Leeuwen, S.P.J. van; Mol, J.G.J. ; Lee, M.K. van der; Gerssen, A. ; Lasaroms, J.J.P. ; Sterk, S.S. ; Raamsdonk, L. aan; Jong, J. de; Scholtens, I.M.J. ; Alewijn, A. ; Silletti, E. ; Ginkel, L. van; Noordam, M.Y. ; Meijer, N. - \ 2018
Wageningen : RIKILT Wageningen University & Research (RIKILT-report 2018.009) - 51
|Experiences with an extended screening for GMO-labelled samples at RIKILT
Scholtens-Toma, Ingrid - \ 2018
|The development of a detection method for poultry processed proteins in feed
Raamsdonk, L.W.D. van; Prins, T.W. ; Scholtens-Toma, I.M.J. - \ 2018
NGS-based amplicon sequencing approach; towards a new era in GMO screening and detection
Arulandhu, Alfred J. ; Dijk, Jeroen van; Staats, Martijn ; Hagelaar, Rico ; Voorhuijzen, Marleen ; Molenaar, Bonnie ; Hoof, Richard van; Li, Rong ; Yang, Litao ; Shi, Jianxin ; Scholtens, Ingrid ; Kok, Esther - \ 2018
Food Control 93 (2018). - ISSN 0956-7135 - p. 201 - 210.
Amplicon sequencing - Bioinformatics - Genetically modified products - Illumina HiSeq - NGS - qPCR - Unauthorized GMOs
The development and commercialization of Genetically Modified Organisms (GMOs) and its related products have been increasing in the last two decades. This challenges the currently applied time-consuming and expensive qPCR screening procedure from a practical perspective, due to the necessity to develop and validate additional targets at a regular pace and the increasing number of targets included in a single screening. In this study we developed a next generation sequencing (NGS)-based GMO screening approach covering 96 GMO targets and compared it to the two-step qPCR GMO screening approach; the two approaches were evaluated with five feed samples known to contain GMOs. The amplicons obtained from the feed samples were analyzed using 150-bp Paired-End sequencing, Illumina HiSeq 4000 platform. A dedicated data analysis pipeline was developed, which allows automated identification of GMOs and associated genetic elements and constructs. The result of the NGS-based screening were compared with the qPCR approach, indicating that 92% of the targets were commonly identified between the qPCR and NGS-based screening. The remaining 8% of the targets had discrepancies in detection between the two methods. This was mainly observed for targets that were detected in qPCR with high Cq values (above 36), which could not be detected in NGS-based screening. Additionally, due to the more extensive screening in the NGS-based strategy, in total 43 additional GMOs and related targets were identified compared to the standard qPCR screening. From the commonly identified targets in both approaches, 8 targets could not be associated with the detected GMOs. These targets had late Cq values (above 36) and could indicate traces of unknown GMOs in the samples. The current study shows the applicability of NGS as a novel, broad and reliable screening strategy for GMOs and its potential to improve current screening methods.
Development and international validation trial of an advanced, multi-locus DNA metabarcoding metho to identify endangered species in complex samples
Arulandhu, A.J. ; Staats, M. ; Hagelaar, Rico ; Voorhuijzen, M.M. ; Prins, T.W. ; Scholtens-Toma, I.M.J. ; Costessi, Adalberto ; Duijsings, Danny ; Rechenmann, François ; Gaspar, Frédéric B. ; Ruth, S.M. van; Kok, E.J. - \ 2017
PRJEB18620 - PRJEB18620 - ERP020564 - CITES ring trial
Background: DNA metabarcoding, which involves Next-Generation Sequencing (NGS) of DNA barcodes, holds great promise for species identification in complex samples such as food supplements and Traditional Medicines (TMs). Such method would aid CITES (the Convention on International Trade in Endangered Species of Wild Fauna and Flora) enforcement officers to combat wildlife crime by preventing illegal trade of endangered plant and animal species. The objective of this research was to develop a multi-locus DNA metabarcoding method for wildlife forensic species identification and to evaluate the applicability and reproducibility of the this approach across different laboratories. Results: The DNA metabarcoding method developed in this study makes use of 12 DNA barcode markers that have demonstrated universal applicability across a wide range of plant and animal taxa, and that facilitate the identification of plant and animal species in highly processed samples containing degraded DNA. The DNA metabarcoding method was developed on the basis of NGS data generated for 15 well-defined experimental mixtures using Illumina MiSeq technology, for which a bioinformatics pipeline with user-friendly web interface was developed. The performance of the DNA metabarcoding method was assessed in an international validation trial by 16 laboratories, in which the method was found to be highly reproducible and sensitive enough to identify species present in a mixture at 1% dry weight content. Conclusion: The advanced, multi-locus DNA metabarcoding method assessed in this study provides reliable and detailed data on the composition of complex food products, including information on the presence of CITES species. The method provides improved resolution for species identification, while verifying species with multiple DNA barcodes contributes to enhanced quality assurance.
De bepaling van diersoorten in vleesmengsels met DNA-gebaseerde qPCR-methoden
Aartse, A. ; Scholtens, I.M.J. ; Kok, E.J. - \ 2017
Wageningen : RIKILT Wageningen University & Research (RIKILT-rapport 2017.009) - 91
Development and validation of a multi-locus DNA metabarcoding method to identify endangered species in complex samples
Arulandhu, Alfred J. ; Staats, Martijn ; Hagelaar, Rico ; Voorhuijzen, Marleen M. ; Prins, Theo W. ; Scholtens, Ingrid ; Costessi, Adalberto ; Duijsings, Danny ; Rechenmann, François ; Gaspar, Frédéric B. ; Barreto Crespo, Maria Teresa ; Holst-Jensen, Arne ; Birck, Matthew ; Burns, Malcolm ; Haynes, Edward ; Hochegger, Rupert ; Klingl, Alexander ; Lundberg, Lisa ; Natale, Chiara ; Niekamp, Hauke ; Perri, Elena ; Barbante, Alessandra ; Rosec, Jean Philippe ; Seyfarth, Ralf ; Sovova, Tereza ; Moorleghem, Christoff Van; Ruth, Saskia van; Peelen, Tamara ; Kok, Esther - \ 2017
GigaScience 6 (2017)10. - ISSN 2047-217X
CITES - COI - Customs agencies - cyt b - DNA metabarcoding - Endangered species - matK - Mini-barcodes - rbcL - Traditional medicines
DNA metabarcoding provides great potential for species identification in complex samples such as food supplements and traditional medicines. Such a method would aid Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) enforcement officers to combat wildlife crime by preventing illegal trade of endangered plant and animal species. The objective of this research was to develop a multi-locus DNA metabarcoding method for forensic wildlife species identification and to evaluate the applicability and reproducibility of this approach across different laboratories. A DNA metabarcoding method was developed that makes use of 12 DNA barcode markers that have demonstrated universal applicability across a wide range of plant and animal taxa and that facilitate the identification of species in samples containing degraded DNA. The DNA metabarcoding method was developed based on Illumina MiSeq amplicon sequencing of well-defined experimental mixtures, for which a bioinformatics pipeline with user-friendly web-interface was developed. The performance of the DNA metabarcoding method was assessed in an international validation trial by 16 laboratories, in which the method was found to be highly reproducible and sensitive enough to identify species present in a mixture at 1% dry weight content. The advanced multi-locus DNA metabarcoding method assessed in this study provides reliable and detailed data on the composition of complex food products, including information on the presence of CITES-listed species. The method can provide improved resolution for species identification, while verifying species with multiple DNA barcodes contributes to an enhanced quality assurance.
China's policies on greening financial institutions: assessment and outlook
Mol, A.P.J. - \ 2017
In: Routledge Handbook of Environmental Policy in China / Sternfeld, E., Routledge - ISBN 9781138831117 - p. 208 - 222.
Environmental protection and sustainability have a complex relationship with finances and financial institutions. Financial institutions such as banks, pension funds and insurance companies, are increasingly seen as of vital importance for reaching environmental and sustainability goals. Initially, and at least till the early 1990s, the availability of finances and the functioning of financial institutions were considered to contribute significantly to unsustainable practices. Through their investment and loan practices, banks and other financial institutions were seen as major drivers behind economic growth and as such major contributors to natural resource depletion and environmental pollution. From the 1990s onwards, the sustainability perspective of financial institutions and the availability of finance has diversified. On the waves of ecological modernization (Spaargaren and Mol 1992) financial institutions were also considered as potentially major institutions, actors and instruments in greening investments, industrial development, and infrastructures. And lifting restrictions on finance was no longer one-to-one related to increasing pollution and resource extraction, but also to greening technological development and capitalization of green transitions (e.g. Scholtens 2006; Perez 2008; Yuxiang and Chen 2011, 95-97; UNEP 2014). Also, there was more and more recognition that public finance alone would never be able to free the finances needed to turn economies and societies green, as major investments will be needed in industrial transformations, new infrastructure, retro-fitting existing buildings, zero-carbon development and the like (Shen et al. 2013). This more positive role of financial institutions for the sustainability agenda started to gain prominence in the scholarly literature in the early 1990s, first especially with respect to international institutions such as the World Bank and its International Financial Corporation, other regional development banks, and the International Monetary Fund (IMF) (Perez 2008). These financial institutions decided - under considerable pressure from NGOs and some states - to integrate ecological considerations and later even conditionality into mainstream lending policies and practices. Following these development banks and international financial institutions, private banks also, starting with the larger ones from OECD countries with international operations, began to subject lending to some form of voluntary environmental regulation. The UNEP Finance Initiative, established in the 1990s, was the first to try to provide a global normative framework for the incorporation of environmental considerations into the business practices of public and private financial institutions. During all these initiatives and related debates it became clear that there were still many teething troubles and conflicting goals, which come along with sustainability roles for finance. The ambivalences of finance, financial instruments and financial institutions for the sustainability agenda made the design and implementation of the former three of key importance, and thus subject of research and politics.
National Reference Laboratories RIKILT Wageningen University & Research : annual report 2016
Leeuwen, S.P.J. van; Mol, J.G.J. ; Lee, M.K. van der; Gerssen, A. ; Lasaroms, J.J.P. ; Sterk, S.S. ; Raamsdonk, L.W.D. van; Jong, J. de; Scholtens-Toma, I.M.J. ; Alewijn, M. ; Weesepoel, Y.J.A. ; Ginkel, L.A. van; Meijer, Nathan ; Noordam, M.Y. - \ 2017
Wageningen : RIKILT Wageningen University & Research (RIKILT Report 2017.007) - 49
reference standards - laboratories - food legislation - europe - annual reports - food safety - food quality - feeding standards - referentienormen - laboratoria - voedingsmiddelenwetgeving - europa - jaarverslagen - voedselveiligheid - voedselkwaliteit - voedingsnormen
National Reference Laboratories (NRLs) are part of the system responsible for the control and enforcement of EU food and feed law. RIKILT Wageningen University & Research has been designated as the NRL for twelve subjects. The tasks of a NRL depend on its research field. This report gives an overview of the activities performed by all of RIKILT's NRLs in 2016.
Evaluation of a loop-mediated isothermal amplification (LAMP) method for rapid on-site detection of horse meat
Aartse, Aafke ; Scholtens-Toma, Ingrid ; A, Hans J.G. van der; Boersma-Greve, Monique M. ; Prins, Theo W. ; Ginkel, Leen A. van; Kok, Esther J. ; Bovee, Toine F.H. - \ 2017
Food Control 81 (2017). - ISSN 0956-7135 - p. 9 - 15.
Horse meat - Loop-mediated isothermal amplification (LAMP) - On-site detection - qPCR - Rapid method
Detection of horse DNA by loop-mediated isothermal amplification (LAMP) seems one of the most promising methods to meet the criteria of fast, robust, cost efficient, specific, and sensitive on-site detection. In the present study an assessment of the specificity and sensitivity of the LAMP horse screening assay was made and outcomes were compared with the EURL-AP (European Union Reference laboratory for Animal Proteins in feeding stuffs) qPCR method. The specificity was tested with DNA samples from seven other species. The sensitivity of the LAMP assay was subsequently challenged with different percentages of horse DNA in cattle DNA and different percentages of horse meat in cattle meat. Both qPCR and LAMP were able to reliably detect horse DNA or meat below 0.1%, but LAMP was able to do so in less than 30 min. The DNA of other species did not result in a response in the LAMP horse assay. These features show that the LAMP method is fast, specific, and sensitive. Next, 69 processed meat samples were screened for the presence of horse DNA. The results showed that the LAMP horse assay, combined with a simple and fast on-site DNA extraction method, results in similar outcomes as the EURL-AP qPCR method and is thus a promising screening assay to be used outside the laboratory. Only samples that are screened on-site as suspect in the LAMP horse assay, need to be brought to the laboratory for confirmation with the more laborious EURL-AP qPCR reference method.
Semiautomated TaqMan PCR screening of GMO labelled samples for (unauthorised) GMOs
Scholtens-Toma, Ingrid ; Molenaar, Bonnie ; Hoof, Richard A. van; Zaaijer, Stephanie ; Prins, Theo W. ; Kok, Esther J. - \ 2017
Analytical and Bioanalytical Chemistry 409 (2017)15. - ISSN 1618-2642 - p. 3877 - 3889.
Automation - Construct - Element - GMO - Screening - Specificity
In most countries, systems are in place to analyse food products for the potential presence of genetically modified organisms (GMOs), to enforce labelling requirements and to screen for the potential presence of unauthorised GMOs. With the growing number of GMOs on the world market, a larger diversity of methods is required for informative analyses. In this paper, the specificity of an extended screening set consisting of 32 screening methods to identify different crop species (endogenous genes) and GMO elements was verified against 59 different GMO reference materials. In addition, a cost- and time-efficient strategy for DNA isolation, screening and identification is presented. A module for semiautomated analysis of the screening results and planning of subsequent event-specific tests for identification has been developed. The Excel-based module contains information on the experimentally verified specificity of the element methods and of the EU authorisation status of the GMO events. If a detected GMO element cannot be explained by any of the events as identified in the same sample, this may indicate the presence of an unknown unauthorised GMO that may not yet have been assessed for its safety for humans, animals or the environment.
Specificity of a novel TaqMan PCR method for detection of poultry DNA
Scholtens-Toma, Ingrid ; Prins, Theo W. ; Raamsdonk, Leo W.D. Van - \ 2017
Food Control 73 (2017). - ISSN 0956-7135 - p. 532 - 539.
After the Bovine Spongiform Encephalopathy (BSE) crisis emerged in 1985/1986, all processed animal proteins (PAPs) were finally banned for use in animal feed in the European Union. To partially lift this feed ban, paths for re-introduction of PAPs from species other than ruminants e.g. pig and poultry, are described in the Transmissible Spongiform Encephalopathies (TSE) Roadmap 2. Cannibalism, however, is still not allowed. Specific detection methods for pig and poultry meal and PAPs are prerequisites for reintroduction of pig and poultry processed animal proteins into animal feed. Developing a sensitive PCR method that specifically detects the taxonomically diverse and therefore artificial group ‘poultry’ and that does not detect other birds at the same time is a challenge. Here, a novel TaqMan PCR method for poultry detection is presented. The specificity of the poultry method against target and non-target species has been extensively investigated. The efficiency, linearity and sensitivity was tested using dilution series of chicken, turkey, duck and goose DNA isolated from meat and autoclaved meat as a model system for PAPs.
Novel TaqMan PCR screening methods for element cry3A and construct gat/T-pinII to support detection of both known and unknown GMOs
Prins, Theo W. ; Hoof, Richard A. van; Scholtens, Ingrid M.J. ; Kok, Esther J. - \ 2017
European Food Research and Technology 243 (2017)3. - ISSN 1438-2377 - p. 481 - 488.
Detection - Identification - PCR - qPCR - Real-time - Screening
The import and use of genetically modified organisms (GMOs) is strictly regulated in the European Union. In order to maintain the legislation on GMOs, a genetic element screening is generally applied as a first step to detect authorised as well as unauthorised GMOs. Subsequent identification of GMOs that relate to the detected elements is performed by the application of event-specific detection methods. However, as the diversity of GMOs on the world market is increasing, there is an ongoing need for methods for additional informative screening elements. Genes that are increasingly applied in GMOs are cry3A (including variants mcry3A and eCry3.1Ab) conferring resistance to Bt toxins, and gat, detoxifying glyphosate. Novel TaqMan PCR detection methods for element cry3A and construct gat/T-pinII were developed to support the identification of maize MIR604, 98140, 5307, canola 61061 and 73496, and soybean 356043. Also, other unknown (unauthorised) GMOs containing cry3A and/or gat/T-pinII can potentially be detected. Specificity, efficiency and sensitivity of the methods were evaluated.
|‘Fishing for Fairness: North Sri Lankan fisher struggles for access to transboundary fishing grounds, South Asia’
Büscher, Bram - \ 2016
Doctoral examination committee: Mr. Joeri Scholtens