6th Meeting of the Advisory Board for Comparative Testing - Financial and Technical Report

In the frame of Regulation (EC) No 882/2004, the European Union Reference Laboratory for Genetically Modified Food and Feed (EU-RL GMFF) has the duty to organise comparative testing rounds for National Reference Laboratories (NRLs). The EU-RL GMFF is assisted in its duties by a so-called Advisory Board for Comparative Testing. The aim of the Advisory Board is to provide support to the organisation of the comparative testing rounds and to give guidance on the approach for participants’ assessment and on participants’ results and performance. Within two months after the Advisory Board meeting has taken place a financial and technical report has to be submitted to DG SANCO. The following topics were discussed at the Advisory Board meeting of 23 and 24 August 2012: - The draft report of the fourth round of comparative testing ILC-EURL-GMFF-CT-02/11 - The standard deviations for proficiency testing of the fifth comparative testing round ILC-EURL-GMFF-CT-01/12 containing oilseed rape event GT73 and maize event 59122 - Preparation of test materials: decision about the test materials for 2013 and brainstorming about future test materialsJRC.I.3-Molecular Biology and Genomic

Event-specific Method for the Quantification of Soybean MON 87701 Using Real-time PCR

The European Union Reference Laboratory for GM Food and Feed (EU-RL GMFF), established by Regulation (EC) No 1829/2003, has carried out a validation study to assess the performance of a quantitative event-specific method on the soybean event MON 87701 (unique identifier MON-877Ø1-2). The collaborative trial was conducted according to internationally accepted guidelines. In accordance to Regulation (EC) No 1829/2003 of 22 September 2003 on genetically modified food and feed and to Regulation (EC) No 641/2004 of 6 April 2004 on detailed rules for the implementation of Regulation (EC) No 1829/2003, Monsanto Company provided the detection method and the control samples. The EU-RL GMFF prepared the validation samples [calibration samples and blind samples at unknown GM percentages (DNA/DNA)]. The results of the international collaborative trial met the European Network of GMO Laboratories (ENGL) method performance requirements (http://gmocrl.jrc.ec.europa.eu/guidancedocs.htm). The method is therefore considered applicable to the control samples provided, in accordance with the requirements of Annex I – 2.C.2 to Regulation (EC) No 641/2004.JRC.I.3-Molecular Biology and Genomic

Certification of Plasmidic DNA containing MON 810 Maize DNA fragments, Certified Reference Materials ERM-AD413

This report describes the preparation, homogeneity, stability and certification studies of a plasmidic deoxyribonucleic acid (DNA) (ERM-AD413) containing a defined DNA fragment specific for a genetic modification present in Zea Mays MON 810 event (1) as well as a defined DNA fragment specific for the Zea Mays taxon. The CRM was processed in 2005 and certified in 2007 by the European Commission, Directorate General Joint Research Centre, Institute for Reference Materials and Measurements (IRMM) in Geel, Belgium. The CRM is available in plastic tubes containing 500 µL of the plasmidic DNA in 1 mmol/L EDTA pH 7.0 buffer. Each tube contains approximately 10 to the power of 9 copies of the ERM-AD413 plasmid which correspond approximately to 3.41 ng of DNA. The plasmid contains a 170 bp fragment of the MON 810 5' plant-P35S junction and a 351 bp fragment of the maize endogenous high mobility group gene (hmg) (2). both DNA sequences originated from DNA extracted from MON 810 seeds supplied by RAGT Semences (Rodez, France). The certified value is the number of each cloned DNA fragments per plasmid. The number ratio between those two DNA fragments is given as an indicative value measured by duplex and simplex real-time PCR.JRC.D.2-Reference material

Comparative Testing Report on the Detection and Quantification of Maize Events GA21, TC1507 and MIR604

In the frame of Regulation (EC) No 882/2004, the European Union Reference Laboratory for Genetically Modified Food and Feed has the duty to organise comparative testing rounds and to ensure an appropriate follow-up of these activities. This preliminary report describes the outcome of the fourth comparative testing round ILC-EURL-GMFF-CT-02/11. Participants were required to screen two test items denoted maize powder levels 1 and 2, for the presence of maize events 3272, Bt11, Bt176, 59122, GA21, MIR604, MON 810, MON 863, NK603 and TC1507. Any events detected were then to be quantified. This comparative testing round was organised in collaboration with the Food Safety and Quality Unit of the Institute for Reference Materials and Measurements (Geel, BE). The maize test items were produced in-house. The Food Safety and Quality Unit managed the on-line registration and submission of results. A total of 159 laboratories were invited to participate in ILC-EURL-GMFF-CT-02/11. Participants could report the results of the exercise either in mass/mass % or in copy/copy %. In this fourth comparative testing round greater than 86 % of participants gained a satisfactory z-score in the range of -2 to +2 for the results expressed in mass/mass % for both maize powder levels 1 and 2 regardless of the GM event. However, a lower percentage (43–86 %) of z-scores within the working range of -2 to +2 was calculated for those participants that expressed the results in copy/copy %.JRC.I.3-Molecular Biology and Genomic

Report on the Verification of the Performance of MON 531 and MON 1445 Event-specific Methods on the Cotton Event MON 531 x MON 1445 Using Real-Time PCR

The JRC as Community Reference Laboratory for GM Food and Feed (CRL-GMFF), established by Regulation (EC) No 1829/2003, has carried out an in-house verification study to assess the performance of two quantitative event-specific methods on the cotton event MON 531 x MON 1445 (unique identifier MON-ØØ531-6 x MON-Ø1445-2) which combines the MON 531 and MON 1445 transformation events. The two methods have been previously validated individually on single-trait events, to detect and quantify each event in cotton samples; a validation report for each method is available at http://gmo-crl.jrc.ec.europa.eu/statusofdoss.htm. This study was conducted according to internationally accepted guidelines (1, 2). In accordance to Regulation (EC) No 1829/2003 of 22 September 2003 on genetically modified food and feed and to Regulation (EC) No 641/2004 of 6 April 2004 on detailed rules for the implementation of Regulation (EC) No 1829/2003, Monsanto Europe S.A. provided the detection methods and the control samples: whole seeds of MON 531 x MON 1445 cotton (ST5599BR) and whole conventional cotton seeds of ST474. The JRC prepared the in-house verification samples (calibration samples and blind samples at different GM percentages). The results of the in-house verification study were evaluated with reference to ENGL method performance requirements (http://gmo-crl.jrc.ec.europa.eu/guidancedocs.htm) and to the validation results on the individual parental events (http://gmo-crl.jrc.ec.europa.eu/statusofdoss.htm). The results of this CRL-GMFF in-house verification studies are made publicly available at http://gmo-crl.jrc.ec.europa.eu/.JRC.DG.I.4 - Molecular biology and genomic

Comparative Testing Report on the Detection and Quantification of Soybean Event 40-3-2 - Comparative testing round: ILC-EURL-GMFF-CT-01/11

In the frame of Regulation (EC) No 882/2004, the European Union Reference Laboratory for Genetically Modified Food and Feed has the duty to organise comparative testing rounds and to ensure an appropriate follow-up of these activities. This report describes the outcome of the third comparative testing round ILC-EURL-GMFF-CT-01/11. Participants had to determine the GM content in two test items denoted soybean powder levels 1 and 2, containing different GM percentages of soybean event 40-3-2 flour. This comparative testing round was organised in collaboration with the Food Safety and Quality Unit of the Institute for Reference Materials and Measurements (Geel, BE). The soybean event 40-3-2 test items were produced in-house. The Food Safety and Quality Unit managed the on-line registration and submission of results. A total of 155 laboratories were invited to participate in ILC-EURL-GMFF-CT-01/11. Eight National Reference Laboratories declined participation, of which one was no longer a National Reference Laboratory. One hundred and two laboratories from 43 countries returned results, of which 62 were National Reference Laboratories, 11 were members of the European Network of GMO Laboratories only, eight were only Official control laboratories and 21 were laboratories from third countries. Seven laboratories including one National Reference Laboratory, one European Network of GMO Laboratory and five laboratories from a third country did not submit results. Participants could report the results of the exercise either in mass/mass % or in copy/copy %. In this third comparative testing round greater than 86 % of participants gained a satisfactory z-score in the range of -2 to +2 for both soybean powder levels 1 and 2 regardless of the calibration method and the measurement unit.JRC.I.3 - Molecular Biology and Genomic

Comparative Testing Report on the Detection and Quantification of Maize Event MON 810 - Comparative testing round: ILC-CRL-GMFF-CT-02/10

In the frame of Regulation (EC) No 882/2004, the European Union Reference Laboratory for Genetically Modified Food and Feed has the duty to organise comparative testing rounds and to ensure an appropriate follow-up of these activities. This report describes the outcome of the second comparative testing round ILC-CRL-GMFF-CT-02/10. Participants had to determine the GM content in two test items denoted maize powder levels 1 and 2, containing different GM percentages of maize event MON 810. This comparative testing round was organised in collaboration with the Reference Materials Unit and the Food Safety and Quality Unit of the Institute for Reference Materials and Measurements (Geel, BE). The maize event MON 810 test items were produced by the Reference Materials Unit. The Food Safety and Quality Unit managed the on-line registration and submission of results. A total of 136 laboratories were invited to participate in ILC-CRL-GMFF-CT-02/10. Six National Reference Laboratories declined participation, of which two were no longer a National Reference Laboratory. Ninety laboratories from 41 countries returned results, of which 65 were National Reference Laboratories, six were members of the European Network of GMO Laboratories only and 19 were laboratories from third countries. Two National Reference Laboratories, two Official control laboratories and nine laboratories from a third country did not submit any results. Participants could report the results of the exercise either in mass/mass % or in copy/copy %. The outcome of this second comparative testing round was in general positive, with 82-100 % of participants gaining a z-score in the range of -2 to +2 for both maize powder levels 1 and 2 regardless of the calibration method, the measurement unit and the approach used for calculating the z-score.JRC.I.3-Molecular Biology and Genomic

Comparative Testing Report on the Detection and Quantification of Maize Event NK603 - Comparative testing round: ILC-CRL-GMFF-CT-01/10

In the frame of Regulation EC (No) 882/2004, the European Reference Laboratory for Genetically Modified Food and Feed has the duty to organise comparative testing rounds and to ensure an appropriate follow-up of these activities. This report describes the outcome of the first comparative testing round ILC-CRL-GMFF-CT-01/10. Participants had to determine the GM content in two test items denoted maize powder level 1 and level 2, containing different GM percentages of maize event NK603. This comparative testing round was organised in collaboration with the Reference Materials Unit and the Food Safety and Quality Unit of the Institute for Reference Materials and Measurements (Geel, BE). The maize event NK603 test items were produced by the Reference Materials Unit. The Food Safety and Quality Unit managed the on-line registration and submission of results. A total of 110 laboratories were invited to participate in ILC-CRL-GMFF-CT-01/10. Five National Reference Laboratories declined participation. Ninety-three results were returned from 84 laboratories from 36 countries, of which 66 were National Reference Laboratories, seven were members of the European Network of GMO Laboratories only and 11 were laboratories from third countries. Two National Reference Laboratories, two European Network of GMO Laboratories only members and one laboratory from a third country did not submit any results. Participants could report the results of the exercise either in mass/mass % or in copy/copy %. The outcome of this first comparative testing round was in general positive with a share of 86-96 % and 94-100 % of participants exhibiting a z-score in the range of -2 to +2 for maize powder level 1 and level 2, respectively.JRC.DG.I.4-Molecular biology and genomic

Guidelines for sample preparation procedures in GMO analysis

This document aims at providing guidelines for a correct sample preparation in GMO analysis on food, feed, seed, plants and propagating material samples. It has been developed by the European Network of GMO Laboratories (ENGL), specifically by the ENGL working group “sample preparation procedures” upon a mandate of the ENGL steering committee. The structure of the document is based on that of an existing ISO standard (ISO 6498:2012), dealing with sample preparation for feed; starting from this, the procedures were adapted to the needs of GMO detection and the scope concerning matrices was widen to food, feed and seeds. These guidelines cover the key steps of the sample preparation, i.e. the size of the laboratory sample, the mass reduction techniques as well as techniques for comminution and mixing; considerations about the test portion are included and some performance tests to be applied at different steps of the process are provided. While sample preparation, falling after sampling and before analytical testing for GMOs, has suffered from a lack of guidance and harmonization, this document provides detailed and practical hints on this important step, thus contributing to foster testing harmonisation.JRC.I.3-Molecular Biology and Genomic