Use of Certified Reference Materials for the Quantification of GMO in DNA Copy Number Ratio

This application note provides guidance on the correct use of European Reference Materials certified for their GM (genetically modified) copy number fraction of a specific GM event. The details given below refer particularly to the use of the CRMs ERM-BF413d and ERM-AD412 and the upcoming CRMs certified for copy number ratio.JRC.D.2-Reference material

ERM®-AD482 and ERM®-AD483: Certified Reference Materials to identify the origin of animal proteins in feed

This leaflet describes ERM®-AD482 and ERM®-AD483.JRC.F.6-Reference Material

Guidance Document on the Use of Reference Materials in Genetic Testing

The aim of this guidance document is to provide a short and user-friendly support to genetic testing laboratories. It is based on internationally harmonised basic concepts and terminology for quality assurance and control (QA/QC). Relevant terms of the International Organization for Standardization (ISO) and the International Vocabulary of Metrology (VIM) are explained and references to other expert texts are provided. The document includes discussion and clarification of several often misunderstood issues. It takes into account the current scientific knowledge and the existing technological capabilities. This document may be revised and constitutes a deliverable of th NoE EuroGentest.JRC.D.2-Reference material

Training manual on GMO quantification: Proper calibration and estimation of measurement uncertainty

The content of this manual is based on the training course that was organised on the premises of the European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (Geel, BE) at the end of 2013. The training manual complements the training course that was intended to improve the quality of measurement results obtained when quantifying genetically modified organisms (GMO) in food and feed. Both, the training course and this manual, were developed in line with the current EU GMO legislation. The manual is addressed to laboratory managers and practitioners in analytical laboratories who perform GM quantification measurements and use reference materials for calibration, quality control and method validation including in-house verification. It is also intended for analysts who need to assess measurement uncertainties as required by (EC) No 1829/2003, (EC) No 619/2011 and ISO/IEC 17025:2005. This training document has been written by JRC-IRMM upon request of the European Union Reference Laboratory for Genetically Modified Food and Feed (EURL-GMFF) to further improve the reporting of National Reference Laboratories (NRLs) nominated under Regulation (EC) No 882/2004 and official GMO control laboratories within the EU. This manual is organised in four chapters covering the proper calibration of PCR methods, the estimation of measurement uncertainty, the establishment of metrological traceability of a measurement result and the way to prove the trueness of measurement results. The training manual is a didactic support of a previous guidance document that outlines issues related to the estimation of measurement uncertainty (MU) in the GMO sector. The training manual is also in line with the European technical guidance document for the flexible scope accreditation of laboratories quantifying GMOs, that is intended for laboratories that are acquiring or are holding a flexible scope of accreditation according to ISO/IEC 17025.JRC.D.2-Standards for Innovation and sustainable Developmen

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

The certification of different mass fractions of MON-Ø4Ø32-6 in soya bean powder Certified Reference Materials ERM®-BF410ap, ERM®-BF410bp, ERM®-BF410cp, ERM®-BF410dp and ERM®-BF410ep

This report describes the production of a set of Certified Reference Materials (CRMs), ERM BF410ap, bp, cp, dp and ep, which are certified for their GTS 40-3-2 soya bean (unique identifier MON-Ø4Ø32-6) event mass fractions. These materials were produced following ISO Guide 34:2009 and are certified in accordance with ISO Guide 35:2006. The materials are intended for the calibration or quality control of real-time PCR measurements to identify GTS 40-3-2 soya bean and quantify its mass fraction. As with any reference material, they can also be used for establishing control charts or for carrying out validation studies. The CRMs were accepted as European Reference Material (ERM®) after peer evaluation by the partners of the European Reference Materials panel.JRC.F.6-Reference Material

Certification of Reference Materials of Cotton Seed Powder with Different Mass Fractions of the Cotton Event T304-40 - Certified Reference Materials ERM®-BF429 (ERM®-BF429a, ERM®-BF429b, ERM®-BF429c)

This report describes the processing and certification of three cotton seed powder Certified Reference Materials (CRMs) containing different mass fractions of genetically modified (GM) T304-40 cotton (ERM-BF429a, b, c). The materials were prepared and certified in 2009/2010 by the European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (IRMM, Geel, Belgium), according to the principles of ISO Guide 34.JRC.DG.D.2-Reference material

Guidance document on Measurement Uncertainty for GMO Testing Laboratories 3rd Edition

This document provides guidance to GMO control laboratories on how to estimate measurement uncertainty (MU) associated with any GMO measurement result. Competent authorities have a responsibility to decide if a measurement result exceeds the legal threshold. Consequently control laboratories must estimate and report the MU of their measurement results. Quantitative polymerase chain reaction (qPCR) and digital polymerase chain reaction (dPCR) are the methods applied for the enforcement of EU law on GMO labelling. This document takes into account: current EU legislation, the availability of certified reference materials (CRMs) and validated methods, and the need for control laboratories which conduct measurements for the enforcement of EU legislation to be accredited according to ISO/IEC 17025. Based on these, two approaches for the estimation of MU for PCR measurement results are presented in this guidance document. References to alternative approaches are also provided.JRC.F.5-Food and Feed Complianc

CERTIFICATION REPORT The certification of different mass fractions of VCO-Ø1981-5 in maize seed powder Certified Reference Materials ERM®-BF438a, ERM®-BF438b, ERM®-BF438c, ERM®-BF438d and ERM®-BF438e

This report describes the production of a set of Certified Reference Materials (CRMs) ERM BF438a, b, c, d and e, which are certified for their VCO-Ø1981-5 mass fractions. These materials were produced following ISO Guide 34:2009 and are certified in accordance with ISO Guide 35:2006. Genetically modified (GM) seeds of the maize event VCO-Ø1981-5 and seeds from a non-GM maize variety were milled to obtain GM and non-GM seed powders with a similar particle size distribution. Mixtures of non-GM and GM maize powder were prepared gravimetrically. The certified value was obtained from the gravimetric preparations, taking into account the genetic purity with respect to the event VCO-Ø1981-5 of the two powder materials and their water mass fractions. The certified values were confirmed by event-specific real-time PCR as an independent verification method (measurements were within the scope of accreditation to ISO/IEC 17025:2005). The uncertainties of the certified values were estimated in accordance with the Guide to the Expression of Uncertainty in Measurement (GUM) [4] and include uncertainties relating to possible inhomogeneity (Section 4), instability (Section 5) and characterisation (Section 6). The materials are intended for the calibration or quality control of real-time PCR measurements to identify VCO-Ø1981-5 maize and/or quantify its mass fraction. As with any reference material, they can also be used for establishing control charts or for carrying out validation studies. The CRMs are available in glass bottles containing at least 1 g of dried maize seed powder, which were sealed under atmosphere of argon. The minimum amount of sample of the mixtures to be used and recommend for the blank and pure VCO-Ø1981-5 materials is 200 mg. The CRMs were accepted as European Reference Material (ERM®) after peer evaluation by the partners of the European Reference Materials consortium.JRC.D.2-Standards for Innovation and sustainable Developmen