XP CEN ISO/TS 15216-2

XP CEN ISO/TS 15216-2

June 2013
Standard Cancelled

Microbiology of food and animal feed - Horizontal method for determination of hepatitis A virus and norovirus in food using real-time RT-PCR - Part 2 : Method for qualitative detection

ISO/TS 15216-2:2013 describes a method for qualitative detection of HAV and NoV genogroups I (GI) and II (GII), from test samples of foodstuffs 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 approach is also relevant for detection of the target viruses on fomites, or of other human viruses in foodstuffs, on food surfaces or on fomites following appropriate validation and using target-specific primer and probe sets.

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Main informations

Collections

National standards and national normative documents

Publication date

June 2013

Number of pages

40 p.

Reference

XP CEN ISO/TS 15216-2

ICS Codes

07.100.30   Food microbiology

Classification index

V08-670-2

Print number

1 - 11/07/2013

International kinship

ISO/TS 15216-2:2013

European kinship

CEN ISO/TS 15216-2:2013
Sumary
Microbiology of food and animal feed - Horizontal method for determination of hepatitis A virus and norovirus in food using real-time RT-PCR - Part 2 : Method for qualitative detection

ISO/TS 15216-2:2013 describes a method for qualitative detection of HAV and NoV genogroups I (GI) and II (GII), from test samples of foodstuffs 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 approach is also relevant for detection of the target viruses on fomites, or of other human viruses in foodstuffs, on food surfaces or on fomites following appropriate validation and using target-specific primer and probe sets.

Standard replaced by (1)
NF EN ISO 15216-2
September 2019
Standard Current
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

<p>This document specifies a method for detection of hepatitis A virus (HAV) and norovirus genogroups I (GI) and II (GII), from test samples of foodstuffs [(soft fruit, leaf, stem and bulb vegetables, bottled water, bivalve molluscan shellfish (BMS)] or surfaces using real-time RT-PCR.</p> <p>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, surfaces or other matrices.</p>

Table of contents
View the extract
  • Avant-propos
    iv
  • Introduction
    vii
  • 1
    Domaine d'application 1
  • 2
    Références normatives 1
  • 3
    Termes et définitions 1
  • 4
    Principe 3
  • 4.1
    Extraction de virus 3
  • 4.2
    Extraction d'ARN 3
  • 4.3
    Transcription inverse suivie d'une réaction de polymérisation en chaîne en temps réel (RT-PCR en temps réel) 4
  • 4.4
    Témoins 4
  • 4.5
    Résultats des essais 4
  • 5
    Réactifs 5
  • 5.1
    Généralités 5
  • 5.2
    Réactifs utilisés dans l'état 5
  • 5.3
    Réactifs préparés 6
  • 6
    Appareillage et matériaux 7
  • 7
    Échantillonnage 8
  • 8
    Mode opératoire 8
  • 8.1
    Exigences de laboratoire générales 8
  • 8.2
    Extraction de virus 8
  • 8.3
    Extraction d'ARN 11
  • 8.4
    RT-PCR en temps réel 11
  • 9
    Interprétation des résultats 13
  • 9.1
    Généralités 13
  • 9.2
    Élaboration des courbes étalon d'ARN viral témoin de processus 13
  • 9.3
    Calcul de l'efficacité de l'amplification 13
  • 9.4
    Calcul du rendement d'extraction 14
  • 9.5
    Limite de détection théorique 14
  • 10
    Expression des résultats 14
  • 11
    Rapport d'essai 15
  • Annexe A (normative) Diagramme de mode opératoire
    16
  • Annexe B (informative) Mélanges maîtres (MasterMix) de la technique RT-PCR en temps réel et paramètres de cycles
    17
  • Annexe C (informative) Amorces et sondes d'hydrolyse de la technique RT-PCR en temps réel pour la détection du VHA, du norovirus GI et du virus Mengo(témoin de processus)
    18
  • Annexe D (informative) Croissance de la souche du virus Mengo MC0 utilisé comme témoin de processus
    20
  • Annexe E (informative) Extraction d'ARN par l'utilisation du système NucliSens. de chez BioMerieux
    21
  • Annexe F (normative) Composition et préparation des réactifs et des tampons
    23
  • Annexe G (informative) Préparations mères d'ARN témoin externe (ARN TE)
    25
  • Annexe H (informative) Disposition de plaque optique type
    27
  • Bibliographie
    28
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