EN ISO 15216-1:2017
(Main)Microbiology of the food chain - Horizontal method for determination of hepatitis A virus and norovirus using real-time RT-PCR - Part 1: Method for quantification (ISO 15216-1:2017)
Microbiology of the food chain - Horizontal method for determination of hepatitis A virus and norovirus using real-time RT-PCR - Part 1: Method for quantification (ISO 15216-1:2017)
ISO 15216-1:2017 specifies a method for the quantification of levels of HAV and norovirus genogroup I (GI) and II (GII) RNA, from test samples of foodstuffs (soft fruit, leaf, stem and bulb vegetables, bottled water, BMS) or food surfaces. Following liberation of viruses from the test sample, viral RNA is then extracted by lysis with guanidine thiocyanate and adsorption on silica. Target sequences within the viral RNA are amplified and detected by real-time RT-PCR.
This method is not validated for detection of the target viruses in other foodstuffs (including multi-component foodstuffs), or any other matrices, nor for the detection of other viruses in foodstuffs, food surfaces or other matrices.
Mikrobiologie der Lebensmittelkette - Horizontales Verfahren zur Bestimmung von Hepatitis A-Virus und Norovirus mittels Real-time-RT-PCR - Teil 1: Verfahren zur Quantifizierung (ISO 15216-1:2017)
Dieses Dokument legt ein Verfahren für die quantitative Bestimmung des Gehalts an RNA von HAV und Noroviren der Genogruppen I (GI) und II (GII) in Untersuchungsproben von Lebensmitteln (weiches Obst, Blatt , Stängel und Zwiebelgemüse, in Flaschen abgefülltes Trinkwasser, zweischalige Weichtiere (Muscheln)) oder Lebensmitteloberflächen fest. Nach der Freisetzung der Viren aus der Untersuchungsprobe wird die virale RNA durch Aufschluss mit Guanidinthiocyanat und Adsorption an Silica( Partikeln) extrahiert. Die Zielsequenzen in der viralen RNA werden amplifiziert und mittels Real time RT PCR detektiert.
Dieses Verfahren ist weder für den Nachweis der Zielviren in anderen Lebensmitteln (einschließlich zusammengesetzter Lebensmittel) oder anderen Matrices noch für den Nachweis anderer Viren in Lebens-mitteln, auf Lebensmitteloberflächen oder anderen Matrices validiert.
Microbiologie dans la chaine alimentaire - Méthode horizontale pour la recherche des virus de l'hépatite A et norovirus par la technique RT-PCR en temps réel - Partie 1: Méthode de quantification (ISO 15216-1:2017)
ISO 15216-1:2017 décrit une méthode de quantification des niveaux d'ARN de VHA et norovirus des génogroupes I (GI) et II (GII) présents dans des échantillons pour essai d'aliments (fruits tendres, légumes feuilles, tiges et bulbes, eau embouteillée, MBV) ou sur des surfaces alimentaires. Après libération des virus contenus dans l'échantillon pour essai, l'ARN viral est extrait par lyse à l'aide de thiocyanate de guanidine et par adsorption sur silice. Les séquences cibles de l'ARN viral sont amplifiées et détectées par la technique RT-PCR en temps réel.
Cette méthode n'est pas validée pour la détection des virus ciblés dans d'autres aliments (y compris les aliments à plusieurs composants) ou d'autres matrices, ni pour la détection d'autres virus dans les aliments, sur les surfaces alimentaires ou dans d'autres matrices.
Mikrobiologija v prehranski verigi - Horizontalna metoda za ugotavljanje virusa hepatitisa A in norovirusov z RT-PCR v realnem času - 1. del: Metoda za kvantifikacijo (ISO 15216-1:2017)
Ta dokument določa kvantitativno metodo za določevanje ravni genske skupine virusa hepatitisa A in norovirusa I (GI) in II (GII) RNK v vzorcih ali živilih (mehkem sadju, listnati in stebelni zelenjavi, čebulnicah, ustekleničeni vodi, BMS) oz. na površinah živil. Po sprostitvi virusov iz vzorca se nato virusni RNK odstrani z lizo z gvanidinijevim tiocianatom in adsorpcijo na silicijev oksid. Ciljna zaporedja v virusni RNK poudari in zazna RT-PCR v realnem času.
Ta metoda ni potrjena za odkrivanje ciljnih virusov v drugih živilih (vključno z večkomponentnimi živili) ali drugih matricah ali odkrivanje drugih virusov v živilih, na površinah živil ali drugih matricah.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-junij-2017
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SIST-TS CEN ISO/TS 15216-1:2013
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Microbiology of the food chain - Horizontal method for determination of hepatitis A virus
and norovirus using real-time RT-PCR - Part 1: Method for quantification (ISO 15216-
1:2017)
Mikrobiologie der Lebensmittelkette - Horizontales Verfahren zur Bestimmung von
Hepatitis A-Virus und Norovirus mittels Real-time-RT-PCR - Teil 1: Verfahren zur
Quantifizierung (ISO 15216-1:2017)
Microbiologie dans la chaine alimentaire - Méthode horizontale pour la recherche des
virus de l'hépatite A et norovirus par la technique RT-PCR en temps réel - Partie 1:
Méthode de quantification (ISO 15216-1:2017)
Ta slovenski standard je istoveten z: EN ISO 15216-1:2017
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN ISO 15216-1
EUROPEAN STANDARD
NORME EUROPÉENNE
March 2017
EUROPÄISCHE NORM
ICS 07.100.30 Supersedes CEN ISO/TS 15216-1:2013
English Version
Microbiology of the food chain - Horizontal method for
determination of hepatitis A virus and norovirus using
real-time RT-PCR - Part 1: Method for quantification (ISO
15216-1:2017)
Microbiologie dans la chaine alimentaire - Méthode Mikrobiologie der Lebensmittelkette - Horizontales
horizontale pour la recherche des virus de l'hépatite A Verfahren zur Bestimmung von Hepatitis A-Virus und
et norovirus par la technique RT-PCR en temps réel - Norovirus mittels Real-time-RT-PCR - Teil 1: Verfahren
Partie 1: Méthode de quantification (ISO 15216- zur Quantifizierung (ISO 15216-1:2017)
1:2017)
This European Standard was approved by CEN on 23 February 2017.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.
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, Serbia, 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
© 2017 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 15216-1:2017 E
worldwide for CEN national Members.
Contents Page
European foreword . 3
European foreword
This document (EN ISO 15216-1:2017) has been prepared by Technical Committee CEN/TC 275 “Food
analysis - Horizontal methods”, the secretariat of which is held by DIN, in collaboration with Technical
Committee ISO/TC 34 "Food products".
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by September 2017 and conflicting national standards
shall be withdrawn at the latest by September 2017.
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.
This document supersedes CEN ISO/TS 15216-1:2013.
This document has been prepared under a mandate given to CEN by the European Commission and the
European Free Trade Association.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the
following countries are bound to implement this European Standard: 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, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and
the United Kingdom.
Endorsement notice
The text of ISO 15216-1:2017 has been approved by CEN as EN ISO 15216-1:2017 without any
modification.
INTERNATIONAL ISO
STANDARD 15216-1
First edition
2017-03
Microbiology of the food chain —
Horizontal method for determination
of hepatitis A virus and norovirus
using real-time RT-PCR —
Part 1:
Method for quantification
Microbiologie dans la chaîne alimentaire — Méthode horizontale
pour la recherche des virus de l’hépatite A et norovirus par la
technique RT-PCR en temps réel —
Partie 1: Méthode de quantification
Reference number
ISO 15216-1:2017(E)
©
ISO 2017
ISO 15216-1:2017(E)
© ISO 2017, Published in Switzerland
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ii © ISO 2017 – All rights reserved
ISO 15216-1:2017(E)
Contents Page
Foreword .v
Introduction .vi
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 3
4.1 Virus extraction . 3
4.2 RNA extraction . 4
4.3 Real-time RT-PCR . 4
4.4 Control materials . 4
4.4.1 Process control virus . 4
4.4.2 Double-stranded DNA (dsDNA) control . 4
4.4.3 EC RNA control . 4
4.5 Test results. 5
5 Reagents . 5
5.1 General . 5
5.2 Reagents used as supplied . 5
5.3 Prepared reagents . 6
6 Equipment and consumables . 7
7 Sampling . 9
8 Procedure. 9
8.1 General laboratory requirements . 9
8.2 Virus extraction . 9
8.2.1 Process control virus material . 9
8.2.2 Negative process control . 9
8.2.3 Food surfaces . 9
8.2.4 Soft fruit, leaf, stem and bulb vegetables . 9
8.2.5 Bottled water .10
8.2.6 Bivalve molluscan shellfish .11
8.3 RNA extraction .11
8.4 Real-time RT-PCR .12
8.4.1 General requirements .12
8.4.2 Real-time RT-PCR analysis .12
9 Interpretation of results .14
9.1 General .14
9.2 Construction of standard curves .14
9.3 Calculation of RT-PCR inhibition .15
9.4 Calculation of extraction efficiency .15
9.5 Sample quantification .16
10 Expression of results .17
11 Precision .
...
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