CEN/TS 16707:2014
(Main)Foodstuffs - Methods of analysis for the detection of genetically modified organisms and derived products - Polymerase chain reaction (PCR) based screening strategies
Foodstuffs - Methods of analysis for the detection of genetically modified organisms and derived products - Polymerase chain reaction (PCR) based screening strategies
This Technical Specification describes screening strategies for the detection of genetically modified (GM) DNA in food products by means of PCR methods. The strategies have been established for food matrices, but it can also be applied to other matrices (e.g. feed, seed and samples from field grown plants).
Detection of GM DNA is based on PCR methods targeting segments of transgenic DNA sequences (genetic elements, genetic constructs or insertion sites of transgenes). Various combinations of these PCR methods are involved in screening strategies. The methods are applied simultaneously or hierarchically. The general strategy is based on the matrix approach. Examples for the implementation and application of this approach are described.
In order to ensure reliable analytical results, the document also provides guidelines for the validation of the performance criteria of qualitative PCR methods applied in screening approaches.
Lebensmittel - Verfahren zum Nachweis von gentechnisch veränderten Organismen und ihren Produkten — Strategien für das Screening mit Polymerase-Kettenreaktion (PCR)
Diese Technische Spezifikation beschreibt Screening-Strategien für den Nachweis von gentechnisch modifizierter (gm) oder gentechnisch veränderter (gv) DNA in Lebensmitteln mittels PCR Verfahren. Die Strategien wurden für Lebensmittelmatrices erarbeitet, können jedoch auch auf andere Matrices angewendet werden (z. B. Futtermittel, Saatgut und Proben von Feldpflanzen).
Der Nachweis von gv-DNA beruht auf PCR Verfahren, die einzelne Segmente transgener DNA Sequenzen (genetische Elemente, genetische Konstrukte oder Insertionsorte von Transgenen) nachweisen. Verschiedene Kombinationen dieser PCR Verfahren werden in die Screening-Strategien einbezogen. Die Verfahren werden gleichzeitig oder hierarchisch angewendet. Die allgemeine Analysestrategie beruht auf dem Matrix-Ansatz. Beispiele für die Implementierung und Anwendung dieses Ansatzes sind beschrieben.
Um zuverlässige Analyseergebnisse sicherzustellen, enthält das vorliegende Dokument außerdem Leitlinien für die Validierung der Leistungskriterien von qualitativen PCR Verfahren, die bei Screening-Ansätzen angewendet werden.
Produits alimentaires - Méthodes d'analyse pour la détection des organismes génétiquement modifiés et des produits dérivés - Stratégies de criblage basées sur l'utilisation de la réaction de polymérisation en chaîne (PCR)
La présente Spécification technique décrit des stratégies de criblage pour la détection d’ADN génétiquement modifié (GM) dans les produits alimentaires au moyen des méthodes de PCR. Ces stratégies ont été élaborées pour les matrices alimentaires mais elles peuvent également être appliquées à d’autres matrices (par exemple aliments pour animaux, graines et échantillons issus de plantes cultivées en plein champ).
La détection d’ADN GM est basée sur des méthodes de PCR ciblant des segments de séquences d’ADN transgéniques (éléments génétiques, construits génétiques ou sites d’insertion de transgènes). Les stratégies de criblage font intervenir différentes combinaisons de ces méthodes de PCR. Ces méthodes sont appliquées simultanément ou de façon hiérarchique. La stratégie générale repose sur l’approche matricielle. Des exemples de mise en oeuvre et d’application de cette approche sont décrits.
Pour garantir des résultats analytiques fiables, le document fournit également des lignes directrices pour la validation des critères de performance des méthodes de PCR qualitative utilisées dans les approches de criblage.
Živila - Analitske metode za odkrivanje gensko spremenjenih organizmov in njihovih produktov - Strategije presejalne analize s polimerazno verižno reakcijo (PCR)
Ta tehnična specifikacija opisuje strategije presejalne analize za odkrivanje gensko spremenjenega (GM) DNK-ja v živilskih proizvodih z metodo PCR. Te strategije so bile pripravljen za živilske matrice, vendar se lahko uporabljajo tudi za druge matrice (npr. za vzorce krme, semen in rastlin, gojenih na polju).
Odkrivanje GM DNK-ja temelji na metodah PCR za iskanje transgenih sekvenc DNK (genskih elementov, genske konstrukte ali mest vstavitve transgenov). V strategije presejalne analize so vključene različne kombinacije teh metod PCR. Metode se uporabljajo hkrati ali hierarhično. Splošna strategija temelji na pristopu matrice. Opisani so primeri implementacije in uporabe tega pristopa.
Za zagotovitev zanesljivih rezultatov analize podaja dokument tudi smernice za preverjanje meril učinkovitosti uporabljenih kvantitativnih metod PCR v pristopih presejalne analize.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
01-december-2014
Živila - Analitske metode za odkrivanje gensko spremenjenih organizmov in
njihovih produktov - Strategije presejalne analize s polimerazno verižno reakcijo
(PCR)
Foodstuffs - Methods of analysis for the detection of genetically modified organisms and
derived products - Polymerase chain reaction (PCR) based screening strategies
Lebensmittel - Verfahren zum Nachweis von gentechnisch modifizierten Organismen und
ihren Produkten - Strategies für das Screening mit Polymerase-Kettenreaktion (PCR)
Produits alimentaires - Méthodes d'analyse pour la détection des organismes
génétiquement modifiés et des produits dérivés - Stratégies de criblage basées sur
l'utilisation de la réaction de polymérisation en chaîne (PCR)
Ta slovenski standard je istoveten z: CEN/TS 16707:2014
ICS:
67.050 Splošne preskusne in General methods of tests and
analizne metode za živilske analysis for food products
proizvode
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
TECHNICAL SPECIFICATION
CEN/TS 16707
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
October 2014
ICS 67.050
English Version
Foodstuffs - Methods of analysis for the detection of genetically
modified organisms and derived products - Polymerase chain
reaction (PCR) based screening strategies
Produits alimentaires - Méthodes d'analyse pour la Lebensmittel - Verfahren zum Nachweis von gentechnisch
détection des organismes génétiquement modifiés et des veränderten Organismen und ihren Produkten - Strategien
produits dérivés - Stratégies de criblage basées sur für das Screening mit Polymerase-Kettenreaktion (PCR)
l'utilisation de la réaction de polymérisation en chaîne
(PCR)
This Technical Specification (CEN/TS) was approved by CEN on 28 June 2014 for provisional application.
The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to submit their
comments, particularly on the question whether the CEN/TS can be converted into a European Standard.
CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS available
promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in parallel to the CEN/TS)
until the final decision about the possible conversion of the CEN/TS into an EN is reached.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2014 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TS 16707:2014 E
worldwide for CEN national Members.
Contents Page
Foreword .3
Introduction .4
1 Scope .5
2 Normative references .5
3 Terms and definitions .5
4 Principle .6
5 Reagents .7
5.1 General .7
5.2 PCR reagents .7
6 Apparatus and equipment .7
7 PCR analysis .7
7.1 General .7
7.2 Screening .7
7.2.1 General .7
7.2.2 Combination of targets .8
7.2.3 Analysis of the output of the first screening .9
7.2.4 Additional screening tests .9
7.3 GM event identification .9
7.3.1 Event specific tests .9
7.3.2 Additional tests . 10
7.4 Interpretation of PCR results . 10
7.4.1 General . 10
7.4.2 Interpretation of results at the limit of detection (LOD) . 10
8 PCR method performance criteria and validation . 11
8.1 General . 11
8.2 Absolute limit of detection (LOD ) . 12
abs
8.3 Specificity and reference materials . 12
8.4 Robustness . 13
8.5 False-positive rate and false-negative rate . 13
8.6 Probability of Detection (POD) . 13
Bibliography . 14
Foreword
This document (CEN/TS 16707:2014) has been prepared by Technical Committee CEN/TC 275 “Food
analysis - Horizontal methods”, the secretariat of which is held by DIN.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to announce this Technical Specification: Austria, Belgium, Bulgaria, Croatia, Cyprus,
Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany,
Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Introduction
Largely, detection of materials derived from genetically modified organisms (GMOs) in a given sample
employs polymerase chain reaction (PCR) analysis, specifically real-time PCR.
A general strategy for GMO detection and identification by means of PCR analysis and a stepwise approach is
described.
In initial screening analysis, DNA sequences of genetic elements common to many GMOs are targeted.
According to its purpose, screening is a test to rapidly and reliably sort samples into groups. Once the
samples are grouped, screening facilitates and potentially reduces subsequent analytical work and results
interpretation. The screening strategy should be adjusted to the scope (food, feed or seed, crop-specific etc.)
of the test(s).
This document takes the general principle of GMO detection strategies as a basis and describes the
underlying analytical steps for complex screening (known as the matrix-approach [2]).
The document is written primarily for screening strategies applying real-time PCR methods. Other PCR
methodologies may be applicable in the same way.
The terms “screening method” and “screening strategies” are not interchangeable and have different
meanings in this document.
1 Scope
This Technical Specification describes screening strategies for the detection of genetically modified (GM) DNA
in food products by means of PCR methods. The strategies have been established for food matrices, but it
can also be applied to other matrices (e.g. feed, seed and samples from field grown plants).
Detection of GM DNA is based on PCR methods targeting segments of transgenic DNA sequences (genetic
elements, genetic constructs or insertion sites of transgenes). Various combinations of these PCR methods
are involved in screening strategies. The methods are applied simultaneously or hierarchically. The general
strategy is based on the matrix approach. Examples for the implementation and application of this approach
are described.
In order to ensure reliable analytical results, the document also provides guidelines for the validation of the
performance of qualitative PCR methods applied in screening approaches.
2 Normative references
The following documents, in whole or in part, are normatively referenced in this document and are
indispensable for its application. For dated references, only the edition cited applies. For undated references,
the latest edition of the referenced document (including any amendments) applies.
EN ISO 21569, Foodstuffs - Methods of analysis for the detection of genetically modified organisms and
derived products - Qualitative nucleic acid based methods (ISO 21569)
EN ISO 21570, Foodstuffs - Methods of analysis for the detection of genetically modified organisms and
derived products - Quantitative nucleic acid based methods (ISO 21570)
EN ISO 21571, Foodstuffs - Methods of analysis for the detection of genetically modified organisms and
derived products - Nucleic acid extraction (ISO 21571)
EN ISO 24276, Foodstuffs - Methods of analysis for the detection of genetically modified organisms and
derived products - General requirements and definitions (ISO 24276)
3 Terms and definitions
For the purposes of this document, the terms and definitions given in EN ISO 24276 and the following apply:
3.1
GMO method matrix
relational presentation (e.g. a table) of symbols or numbers
Note 1 to entry: One dimension (e.g. columns) corresponds to genetic elements and genetic constructs detected by a
defined PCR method and the other dimension (e.g. rows) corresponds to GM events. The entered symbols or numbers
indicate the detectability or non-detectability of the target sequence for the GM event.
Note 2 to entry: The term matrix is commonly used for a defined composition of food, but this definition is not relevant
here.
3.2
GMO target matrix
relational presentation (e.g. a table) of symbols or numbers
Note 1 to entry: One dimension corresponds to genetic elements or genetic constructs present in a GMO and the
other dimension (e.g. rows) corresponds to GM events. The entered symbols or numbers indicate the presence or
absence of the target for the GM event and copy number, if available.
Note 2 to entry: In contrast to GMO method matrix, the GMO target matrix is independent from a detection method.
3.3
screening method
method that rapidly and reliably eliminates (screens) a large number of negative (or positive) test samples and
restricts the number of test samples requiring the application of a rigorous method
Note 1 to entry: In this document, a screening method refers to PCR methods detecting the presence of several
GMOs in one test.
3.4
element-specific method
method that targets a single discrete DNA sequence of a specific genetic element
Note 1 to entry: A genetic element is a part of a gene, for example a promoter, terminator, intron or a coding
sequence. Elements are often derived from naturally occurring viruses, bacteria, plants, etc. However, elements in GMOs
are commonly modified at the sequence level, relative to the original (natural) source, e.g. by altered codon-usage or
specific single nucleotide polymorphisms (SNPs). This sequence modification may be based on adaptation of the
nucleotide compo
...
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