SIST-TP CEN/TR 16338:2012
(Main)Foodstuffs - Detection of food allergens - Template for supplying information about immunological methods and molecular biology methods
Foodstuffs - Detection of food allergens - Template for supplying information about immunological methods and molecular biology methods
This Technical Report describes necessary information for method providers which needs to be provided with proposals for new work items for consideration in CEN/TC 275/WG 12 "Food allergens".
Lebensmittel - Nachweis von Lebensmittelallergenen - Vorlage zur Bereitstellung von Informationen über immunologische und molekularbiologische Verfahren
Produits alimentaires - Élément central - Élément complémentaire
Le présent rapport technique, fait état des informations nécessaires aux fournisseurs de méthode qui ont besoin d’être fournies avec toute proposition de nouveau sujet au groupe de travail CEN/TC 275/GT 12 « Allergènes alimentaires ».
Živila - Odkrivanje prisotnosti alergenov - Predloga za zagotavljanje informacij o imunoloških in molekularno bioloških metodah
To tehnični poročilo opisuje potrebne informacije, ki jih morajo ponudniki metod predložiti s predlogi za nove elemente dela za obravnavo v standardu CEN/TC 275/WG 12 »alergeni v živilih«.
General Information
Standards Content (Sample)
SLOVENSKI STANDARD
SIST-TP CEN/TR 16338:2012
01-september-2012
Živila - Odkrivanje prisotnosti alergenov - Predloga za zagotavljanje informacij o
imunoloških in molekularno bioloških metodah
Foodstuffs - Detection of food allergens - Template for supplying information about
immunological methods and molecular biology methods
Lebensmittel - Nachweis von Lebensmittelallergenen - Vorlage zur Bereitstellung von
Informationen über immunologische und molekularbiologische Verfahren
Produits alimentaires - Élément central - Élément complémentaire
Ta slovenski standard je istoveten z: CEN/TR 16338:2012
ICS:
67.050 Splošne preskusne in General methods of tests and
analizne metode za živilske analysis for food products
proizvode
SIST-TP CEN/TR 16338:2012 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST-TP CEN/TR 16338:2012
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SIST-TP CEN/TR 16338:2012
TECHNICAL REPORT
CEN/TR 16338
RAPPORT TECHNIQUE
TECHNISCHER BERICHT
May 2012
ICS 67.050
English Version
Foodstuffs - Detection of food allergens - Template for supplying
information about immunological methods and molecular
biological methods
Produits alimentaires - Détection des allergènes Lebensmittel - Nachweis von Lebensmittelallergenen -
alimentaires - Modèle pour la mise à disposition Vorlage zur Bereitstellung von Informationen über
d'informations sur les méthodes d'analyse immunologique immunologische und molekularbiologische Verfahren
et les méthodes d'analyse de biologie moléculaire
This Technical Report was approved by CEN on 4 March 2012. It has been drawn up by the Technical Committee CEN/TC 275.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, 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
Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2012 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN/TR 16338:2012: E
worldwide for CEN national Members.
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Contents Page
Foreword .3
1 Scope .3
2 Normative references .3
3 Terms, definitions and abbreviations .3
4 Necessary elements for method applications .3
5 Estimation of measurement uncertainty .3
6 Physical/environmental conditions .4
7 Instrument calibration .4
8 Safety precautions/safety measures .4
9 Pollution prevention/waste disposal .4
10 Appendices .4
Annex A (informative) General elements for immunological method proposals .5
A.1 Title .5
A.2 Purpose, relevance and scientific basis .5
A.3 Scope .5
A.4 Test principle/summary and analysis steps for immunological methods .5
A.5 Validation status and performance criteria/method performance for immunological
methods .7
Annex B (informative) General elements for molecular biological method proposals . 10
B.1 Title . 10
B.2 Purpose, relevance and scientific basis . 10
B.3 Scope . 10
B.4 Test principle/summary and analysis steps for molecular biological methods . 10
B.5 Validation status and performance criteria for molecular biological methods . 12
Bibliography . 15
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Foreword
This document (CEN/TR 16338:2012) 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.
1 Scope
This Technical Report describes necessary information for method providers which needs to be provided with
proposals for new work items for consideration in CEN/TC 275/WG 12 "Food allergens".
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 15633-1:2009, Foodstuffs — Detection of food allergens by immunological methods — Part 1: General
considerations
EN 15634-1:2009, Foodstuffs — Detection of food allergens by molecular biological methods — Part 1:
General considerations.
EN 15842:2010, Foodstuffs — Detection of food allergens — General considerations and validation of
methods
ISO 5725-1:1994, Accuracy (trueness and precision) of measurement methods and results — Part 1: General
principles and definitions
3 Terms, definitions and abbreviations
For the purpose of this document, the terms and definitions given in EN 15633-1:2009, EN 15634-1:2009 and
EN 15842:2010 apply.
4 Necessary elements for method applications
Each method should include:
a) general information (e. g. title, purpose, relevance and scientific basis, scope and test principle);
b) a detailed description of the method (e. g. reagents, equipment, procedure, calculations, reporting);
c) validation and performance criteria.
All necessary elements for immunological and molecular biological methods are given in Annex A and Annex
B respectively.
5 Estimation of measurement uncertainty
Uncertainty arises from many sources, including the size of the laboratory sample, sampling of the test sample
from the laboratory sample, measurement of the allergen concentration in the extracts, etc. An estimate of the
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measurement uncertainty for each matrix in the area of application will be estimated either from intra/inter-
laboratory study data, or from estimates of the components as described by ISO/IEC Guide 98-3:2008 [1] and
ISO 5725-2:1994 [2].
6 Physical/environmental conditions
Specify conditions (if there are special environmental conditions for performing the analysis), such as normal
laboratory conditions, or use of a cold room, performance of certain steps at specific temperatures etc.
7 Instrument calibration
Instruments should be calibrated as specified in EN ISO/IEC 17025:2005 [3].
8 Safety precautions/safety measures
Describe any particular safety measures (not including country/region specific issues) that shall be brought to
the attention of the analyst.
9 Pollution prevention/waste disposal
The best, most appropriate local practices shall be adhered to.
10 Appendices
The method shall include such diagrams and tables as are necessary for use by the analyst as informative
data from the internal validation study and informative data from the collaborative trial.
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Annex A
(informative)
General elements for immunological method proposals
A.1 Title
The title of the method should include an indication of the species and event or sequence to be detected.
It is suggested that the name and address of the person or institution submitting the method also be included
in the following manner: author submitting company and independent laboratories (if participants). This
information however shall not appear in the published document.
A.2 Purpose, relevance and scientific basis
A description on the purpose of the assay and the scientific background shall be written.
A.3 Scope
A description of the parameters shall be stated in the methods prepared to be a CEN Technical Specification
for the immunological measurements of allergens in food. The allergen and matrix on which the method has
been validated shall be described.
A.4 Test principle/summary and analysis steps for immunological methods
A.4.1 General
A.4.1.1 Terms and definitions
For the purposes of this document, the terms and definitions given in EN 15842:2010 and EN 15633-1:2009
apply.
A.4.1.2 Description of method
The area of application shall be defined. A short description of the method including the method principle shall
be given.
EXAMPLE State which antibodies are used to determine the analyte. Include the specificity of the antibodies
(polyclonal, monoclonal, recombinant, etc.). Describe against which source the antibodies are raised (defined allergen,
protein, marker to be specified. Finally, state the immunoassay format used.
It is mandatory to describe the design of the assay as well as a detailed description of the used antibodies
(including information regarding their purification, characterised affinity and against which substances they are
raised). References to relevant scientific publications are also desirable inclusions.
A.4.1.3 Sample type and matrices
The material used for test calibration shall be given (i.e. whether it is whole extract, non-purified, purified,
fractionated, etc.). Give also a short description of the type of samples and matrices upon which the method
has been validated and can be applied.
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A.4.1.4 Interferences
List conditions and materials, which are known to interfere in both a positive and a negative fashion with the
method, such as samples derived from matrices that interfere with the method.
A.4.2 Apparatus and equipment for immunological methods
All the specific equipment used, including for example spectrophotometers, blenders or other grinding
apparatus, sieves, apparatus or equipment for measuring concentrations (but excluding general laboratory
disposables and equipment), shall be stated. Examples for sources of equipments and reagents including the
relevant disclaimer/non-endorsement should be stated as footnotes.
A.4.3 Reagents (supplies)
A.4.3.1 General
List all the reagents needed. Include at least all those described in EN 15633-1:2009 and all additionally
specified reagents. This listing shall also include all used buffers for assay completion, used reagents and
solutions used for sample treatment. The quality of the reagents and the specific reagents used shall be
indicated. All ingredients and components (for example buffers, including the chemical composition of buffer),
shall be listed.
A.4.3.2 Reagents preparation
If some reagents need to be handled before use (e. g. diluted), the general preparation guidelines shall be
given.
A.4.4 Analysis steps
A.4.4.1 General
In this Clause, give a detailed description of each step of the analysis, from sample preparation to the
immunoassay procedure, so that a trained analyst can perform the method, including all steps involved.
A.4.4.2 General instructions/recommendations
If there are general recommendations to assure a good test performance, this shall be stated (e. g. at which
temperatures the reagents should be used).
A.4.4.3 Preparation of sample for immunological methods
A.4.4.3.1 Sample type and amounts, including sample identification
Give details of the type of laboratory and test samples required (analyte and matrix), with particular reference
to issues of sampling. All samples shall be identified unambiguously.
A.4.4.3.2 Sample collection, transport, preservation and storage
Describe any provisions regarding sample collection and sampling, as well as storage conditions.
A.4.4.3.3 Test sample preparation
Outline the steps of the test sample preparation. Include such details as the
grinding and sieving steps,
sample amount to be weighed in,
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additional reagents,
amount, kind and composition of buffer,
extraction time and temperature.
Describe the method used to extract allergens from the matrix; if applicable use the method from the kit insert.
A.4.4.4 Immunoassay procedure/operational scheme
Describe the procedure in sufficient detail that a trained analyst can perform the method, including any special
steps involved.
A.4.5 Reading/interpretation and test result report (calculations, reporting) for
immunological methods
A.4.5.1 Identification
If the result of the analysis can be verified, indicate how this can be achieved.
A.4.5.2 Calculations
Describe any calculations or mathematical models used to derive the analytical result. Recommendations for
the best curve-fitting model shall be described (e. g. 4-parameter, cubic spline or linear regression model) for
the standard curve (method dependent). Describe any calculations or models used to derive the analytical
result, including conversion factors from, for example, specific of total protein amount to the amount of
allergenic component.
A.4.5.3 Acceptance/rejection criteria
A validated method includes criteria from which an observed measurement result can be accepted as valid.
Describe the acceptance/rejection criteria for the analysis.
A.4.5.4 Reporting
Record keeping should be in conformity with EN ISO/IEC 17025:2005 [3]. Describe how data should be
reported. Normally the mathematical models can be calculated from the usual software delivered with each
ELISA-reader. If no software is available for calculations, alternatives shall be given (e. g. semi-logarithmic
mm-paper, x-axis conc. Standard mg/kg, y-axis ABS standard).
A.5 Validation status and performance criteria/method performance for
immunological methods
A.5.1 General
A summary of the validated performance claims as well as the following the data collected from the internal
validation (including precision, sensitivity, accuracy, specificity and ruggedness), shall be given. If a
collaborative trial was also undertaken, information about such a trial (how many laboratories participated,
outlier elimination, mean values, repeatability r, S , RSD , reproducibility R, S , RSD , etc.), shall be given.
r r R R
A.5.2 Internal validation (manufacturer´s in house study)
Give information obtained from the intra-laboratory trial, including the RSD .
r
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A.5.2.1 Precision (intra- and inter-assay variation)
A.5.2.1.1 Intra-assay variation
Standards shall be measured from a sufficient number of repetitions. Standard deviations and intra-assay CVs
(coefficient of variation) should be given, including the mean CV. Spiked samples can also be measured.
A.5.2.1.2 Inter-assay variation
For the inter-assay variation, standards shall be measured in duplicates in a minimum of six independent runs.
The mean CVs should be calculated. Spiked samples can also be measured.
A.5.2.2 Sensitivity
A.5.2.2.1 Limit of detection (LOD) (range of application)
The detection limit
...
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