EN ISO 15216-2:2019

SLOVENSKI STANDARD
01-december-2019
Nadomešča:
SIST-TS CEN ISO/TS 15216-2:2013
Mikrobiologija v prehranski verigi - Horizontalna metoda za ugotavljanje virusa
hepatitisa A in norovirusov z RT-PCR v realnem času - 2. del: Metoda za
ugotavljanje (ISO 15216-2:2019)
Microbiology of the food chain - Horizontal method for determination of hepatitis A virus
and norovirus using real-time RT-PCR - Part 2: Method for detection (ISO 15216-2:2019)
Mikrobiologie der Lebensmittelkette - Horizontales Verfahren zur Bestimmung von
Hepatitis A-Virus und Norovirus in Lebensmitteln mittels Real-time-RT-PCR - Teil 2:
Nachweisverfahren (ISO 15216-2:2019)
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 2:
Méthode de détection (ISO 15216-2:2019)
Ta slovenski standard je istoveten z: EN ISO 15216-2:2019
ICS:
07.100.30 Mikrobiologija živil Food microbiology
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.

EN ISO 15216-2
EUROPEAN STANDARD
NORME EUROPÉENNE
September 2019
EUROPÄISCHE NORM
ICS 07.100.30 Supersedes CEN ISO/TS 15216-2:2013
English Version
Microbiology of the food chain - Horizontal method for
determination of hepatitis A virus and norovirus using
real-time RT-PCR - Part 2: Method for detection (ISO
15216-2:2019)
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 in Lebensmitteln mittels Real-time-RT-PCR -
Partie 2: Méthode de détection (ISO 15216-2:2019) Teil 2: Nachweisverfahren (ISO 15216-2:2019)
This European Standard was approved by CEN on 27 July 2019.

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, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, 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: Rue de la Science 23, B-1040 Brussels
© 2019 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN ISO 15216-2:2019 E
worldwide for CEN national Members.

Contents Page
European foreword . 3

European foreword
This document (EN ISO 15216-2:2019) has been prepared by Technical Committee ISO/TC 34 "Food
products" in collaboration with Technical Committee CEN/TC 275 “Food analysis - Horizontal methods”
the secretariat of which is held by DIN.
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 March 2020, and conflicting national standards shall
be withdrawn at the latest by March 2020.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document supersedes CEN ISO/TS 15216-2:2013.
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, France, Germany, Greece, Hungary, Iceland,
Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of
North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the
United Kingdom.
Endorsement notice
The text of ISO 15216-2:2019 has been approved by CEN as EN ISO 15216-2:2019 without any
modification.
INTERNATIONAL ISO
STANDARD 15216-2
First edition
2019-07
Microbiology of the food chain —
Horizontal method for determination
of hepatitis A virus and norovirus
using real-time RT-PCR —
Part 2:
Method for detection
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 2: Méthode de détection
Reference number
ISO 15216-2:2019(E)
©
ISO 2019
ISO 15216-2:2019(E)
© ISO 2019
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting
on the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address
below or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Fax: +41 22 749 09 47
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii © ISO 2019 – All rights reserved

ISO 15216-2:2019(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 . 3
4.3 Real-time RT-PCR . 3
4.4 Control materials . 4
4.4.1 Process control virus . 4
4.4.2 EC RNA control . 4
4.5 Test results. 4
5 Reagents . 4
5.1 General . 4
5.2 Reagents used as supplied . 4
5.3 Reagents requiring preparation . 6
6 Equipment and consumables . 7
7 Sampling . 8
8 Procedure. 8
8.1 General laboratory requirements . 8
8.2 Virus extraction . 8
8.2.1 General. 8
8.2.2 Process control virus material . 9
8.2.3 Negative process control . 9
8.2.4 Surfaces . 9
8.2.5 Soft fruit and leaf, stem and bulb vegetables. 9
8.2.6 Bottled water .10
8.2.7 Bivalve molluscan shellfish (BMS) .10
8.3 RNA extraction .11
8.4 Real-time RT-PCR .11
8.4.1 General requirements .11
8.4.2 Real-time RT-PCR analysis .12
9 Interpretation of results .14
9.1 General .14
9.2 Construction of process control virus RNA standard curve.14
9.3 Control for RT-PCR inhibition.14
9.4 Calculation of extraction efficiency .15
10 Expression of results .15
11 Performance characteristics of the method .16
11.1 Validation study.16
11.2 Sens
...