TRAINING COURSE ON USER MANUAL. Edited by Maddalena Querci, Marco Jermini and Guy Van den Eede

Transcription

1 TRAINING COURSE ON THE ANALYSIS OF FOOD SAMPLES FOR THE PRESENCE OF GENETICALLY MODIFIED ORGANISMS USER MANUAL Edited by Maddalena Querci, Marco Jermini and Guy Van den Eede This publication is also available online at: ISBN: Catalogue number: LB-X EN-C Edition 2006 WORLD HEALTH ORGANIZATION REGIONAL OFFICE FOR EUROPE ORGANISATION MONDIALE DE LA SANTE BUREAU REGIONAL DE L'EUROPE WELTGESUNDHEITSORGANISATION REGIONALBÜRO FÜR EUROPA ВСЕМИРНАЯ ОРГАНИЗАЦИЯ ЗДРАВООХРАНЕНИЯ ЕВРОПЕЙСКОЕ РЕГИОНАЛЬНОЕ БЮРО

2 II LEGAL NOTICE Neither the European Commission nor any person acting on behalf of the Commission is responsible for the use which might be made of this publication. A great deal of additional information on the European Union is available on the Internet. It can be accessed through the Europa server Luxembourg: Office for Official Publications of the European Communities ISBN European Communities, 2006 Reproduction is authorised provided the source is acknowledged Printed in Italy

3 III EDITORS Maddalena QUERCI European Commission Joint Research Centre Institute for Health and Consumer Protection Molecular Biology and Genomics Unit Project Manager Marco JERMINI Repubblica e Cantone Ticino Dipartimento della sanità e della socialità Divisione della salute pubblica Laboratorio Cantonale - Bellinzona marco.jermini@ti.ch Guy VAN DEN EEDE European Commission Joint Research Centre Institute for Health and Consumer Protection Molecular Biology and Genomics Unit Head of Unit JRC-BGMO@ec.europa.eu

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5 V Foreword The Institute for Health and Consumer Protection of the Joint Research Centre of the European Commission and the Food Safety Programme within the European Centre for Environment and Health - Rome Division (ECR) of the World Health Organization have jointly organised a series of training courses on The Analysis of Food Samples for the Presence of Genetically Modified Organisms. The Joint Research Centre gives scientific and technical support to EU policies by collaborating with EC Directorates General and by interacting with European Institutions, Organizations and Industries through networking with Member State laboratories. The overall task of the WHO s ECR is to provide support in a complete and coordinated way to both decision-makers and to European citizens in the environmental health field. These training courses are part of collaboration between both Institutions to promote food safety related issues in the WHO European Region, within and beyond actual EU borders, taking into special consideration EU Accession Countries, as well as Central and Eastern Countries with transitional economies. The scope of the training courses is to assist staff of control laboratories to become accustomed with molecular detection techniques, and to help them adapt their facilities and work programmes to include analyses that comply with worldwide regulatory acts in the field of biotechnology. The courses are intended to teach molecular detection techniques to laboratory personnel with a good level of analytical knowledge, but with no or little expertise in this specific domain. The Joint Research Centre has been committed to providing training in detection and quantification of GMOs and, besides the training courses, it offers, and has offered in the past, individual training for specific needs. Training in this topic has been frequently requested due to its importance according to the increasing need to comply with the current and developing European legislative framework. Over the years, the Molecular Biology & Genomics Unit has developed a profound knowledge of the different aspects related to GMO detection and quantification, and

6 has designed, adapted or validated advanced methods for their detection and quantification. VI Knowledge of these techniques has been transferred to collaborating laboratories through publications, collaborative projects, individual training or specific courses. Technical details have also been provided to trainees as oral presentations or brief written outlines. Aware of the need for a permanent source of information, the Molecular Biology & Genomics Unit staff developed this manual, which describes some of the techniques used in our laboratory. The following areas are covered throughout the courses; - DNA extraction from raw and processed materials - Screening of foodstuffs for the presence of GMOs by simple Polymerase Chain Reaction and by nested Polymerase Chain Reaction - Quantification of GMOs in ingredients by real-time Polymerase Chain Reaction - Quantification of GMOs in ingredients by the Enzyme-Linked ImmunoSorbent Assay This Manual has been prepared at the Joint Research Centre, Institute for Health and Consumer Protection (IHCP) as background information for course participants and is intended to provide the theoretical and practical information on methodologies and protocols currently used. The subject matter covers a wide variety of techniques for GMOs detection, identification, characterisation, and quantification, and includes theoretical information considered important background information for anyone wishing to enter and work in the field of GMO detection. It is our hope that the structure and content of this manual will help course participants (as well as other users) in the diffusion and dissemination of the acquired skills in the context of the different working environments according to needs.

7 VII In no way was there an attempt to compete with information available in textbooks or journals. This manual aims to complement existing information in the specialised literature. To facilitate diffusion and consultation, this publication is also available online at: JRC staff members who participated in the preparation of this manual were supervised by Maddalena Querci and are mentioned in the Table of Content according to their contribution. A special recognition and acknowledgment is hereby also given to all Unit personnel who, even not individually mentioned, contributed to the successful preparation of the manual. Thanks are also extended to Sabrina Miglierina, Elisabeth Dilger, Manuela Zingales, Stephen Langrell and Steven Price for their support and collaboration for the revision of this manual. Maddalena Querci, PhD Course co-ordinator June 2004

8 VIII Introductory Remarks from the World Health Organization The World Health Organization gives high priority to the safe use and application of modern biotechnology to food production and processing. For this reason WHO has been very active in the organization of expert consultations on the safety of foods derived from biotechnology since early 90 s. In May 2000, the 53rd World Health Assembly adopted a resolution that WHO should support Member States to provide scientific basis for health-related decision regarding genetically modified foods. Additionally, the WHO Executive Board envisaged that other relevant considerations would be explored in collaboration with other agencies. The Codex Alimentarius Commission (CAC) is an intergovernmental body set up to establish international standards on foods. Its primary objective is to protect the health of consumers and to ensure fair practices in food trade. WHO has been one of the parent organizations of the CAC since its establishment in Within the framework of the Codex Alimentarius activity WHO, and its UN sister Organization FAO, have been involved in the Ad Hoc Intergovernmental Codex Task Force on Foods Derived from Biotechnology, the Codex Committee on Food Labeling and the Codex Committee for Methods of Analysis and Sampling. The WHO Food Safety Programme in Europe has been collaborating with the Joint Research Centre (JRC) of the European Commission since 2000 in the organization of training courses on detection techniques for Genetically Modified Organisms (GMOs) in foods. The aim of these trainings is to provide analytical biotechnology skills to food control laboratory staff and promote the use of validated and harmonized methods for detecting GMOs in Europe and internationally. Adequate methods for analysis and sampling are essential for the appropriate labeling of foods to increase transparency on production processes and to facilitate traceability thus contributing to strengthen food safety systems. To allow broader access to these methods it has been decided that the tutorial manual used during the JRC/WHO Joint Training Courses will be published on the web. The methods presented in this manual are in line with those considered by the Codex Committee for Methods of Analysis and Sampling.

9 IX The WHO Food Safety Programme in Europe recognizes the importance of the collaboration between the European Commission Joint Research Centre, a worldwide recognized scientific institution in the field of GMOs, and will continue promoting capacity building activities in the field of detection methods of GMOs in foods in Europe and other Regions. Cristina Tirado, DVM, PhD WHO Food Safety Regional Adviser in Europe

10 X TRAINING COURSE ON: The Analysis of Food Samples for the Presence of Genetically Modified Organisms USER MANUAL Edited by Maddalena Querci, Marco Jermini and Guy Van den Eede Table of contents Session Title Authors 1 Overview, general introduction on Genetically Modified Organisms (GMOs), EU legislation 2 Manual presentation, working methods and course introduction M. Querci, G. Van den Eede, M. Jermini M. Querci 3 Samples used during the course M. Querci, N. Foti 4 Extraction and purification of DNA M. Somma 5 Agarose gel electrophoresis M. Somma, M. Querci 6 The Polymerase Chain Reaction (PCR) M. Somma, M. Querci 7 Characteristics of Roundup Ready soybean, MON810 maize, and Bt-176 maize 8 Characteristics of the qualitative PCR systems described in the manual 9 Qualitative detection of MON810 maize, Bt-176 maize and Roundup Ready soybean by PCR M. Querci, M. Mazzara M. Querci, M. Mazzara M. Querci, M. Maretti, M. Mazzara 10 Quantitative PCR for the detection of GMOs F. Weighardt 11 Quantitative detection of Roundup Ready soybean by real-time PCR 12 Quantitative detection of Roundup Ready soybean by ELISA Appendix Example of work programme N. Foti F. Eyquem M. Querci