ISO 15216-1:2017 PDF

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

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Title (English):

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

Document status:: Active
Format:: Electronic (PDF)
Number of pages:: 48
Publication date:: March 9, 2017
Edition:: ISO IS 15216 edition 1 version 1
ICS:: 07.100.30

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Description (English):

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.

Technical details

Technical committee: ISO/TC 34/SC 9 - Microbiology
SKU: ISO 15216-1:2017

Abstract

Overview

ISO 15216-1:2017 defines a horizontal method for quantification of hepatitis A virus (HAV) and norovirus genogroups I (GI) and II (GII) in selected food and food-related matrices using real-time RT‑PCR. The standard covers sample-specific virus liberation procedures (e.g., soft fruit, leafy/stem/bulb vegetables, bottled water, bivalve molluscan shellfish (BMS) and food surfaces), RNA extraction by lysis with guanidine thiocyanate and adsorption on silica, and target detection by hydrolysis-probe real-time RT‑PCR to produce quantitative viral RNA results.

Key topics and technical requirements

Scope and limitations: Quantification of HAV and norovirus GI/GII RNA in specified matrices. Not validated for other foodstuffs (including multi-component foods), other matrices or other viruses.
Sample processing: Matrix-specific virus liberation (swabbing for surfaces; PEG/NaCl elution for soft fruit and vegetables; positively charged membranes and ultrafiltration for bottled water; proteinase K for BMS).
RNA extraction method: Chaotropic lysis (guanidine thiocyanate) followed by RNA adsorption onto silica particles to remove RT‑PCR inhibitors.
Real-time RT‑PCR: Amplification and detection with hydrolysis probes for sensitive, specific quantification and confirmation during amplification.
Controls and calibration:
- Process control virus (e.g., mengo virus) to monitor extraction and processing.
- Linear dsDNA control for quantification standards.
- External control (EC) RNA and negative controls to assess inhibition and contamination.
Data interpretation: Construction of standard curves, calculation of extraction efficiency, assessment of RT‑PCR inhibition, and expression of results with defined precision and reproducibility data.
Quality requirements: Laboratory practices, equipment, reagent preparation, plate layouts and validation guidance (see annexes).

Applications and users

ISO 15216-1:2017 is intended for:

Food testing laboratories performing viral contamination testing of soft fruit, vegetables, bottled water, BMS and food-contact surfaces.
Public health agencies and food safety authorities needing standardized quantitative viral RNA data for outbreak investigation and surveillance.
Industry quality teams and third-party testing services requiring harmonized methods for HAV and norovirus monitoring and risk assessment.

Practical benefits include consistent quantification of viral RNA, improved comparability of results between labs, and a structured approach to control for RT‑PCR inhibition and extraction efficiency.

Related standards

ISO 15216 series (other parts and validation guidance)
ISO 22119, ISO 22174, ISO 20838 (PCR/real-time PCR and microbiology of food) For authoritative details and annex content (primers, mastermixes, validation data), consult the full ISO 15216-1:2017 document on the ISO website.

Related standards

Standards mentioned in the description

Abstract

Overview

Key topics and technical requirements

Scope and limitations: Quantification of HAV and norovirus GI/GII RNA in specified matrices. Not validated for other foodstuffs (including multi-component foods), other matrices or other viruses.
Sample processing: Matrix-specific virus liberation (swabbing for surfaces; PEG/NaCl elution for soft fruit and vegetables; positively charged membranes and ultrafiltration for bottled water; proteinase K for BMS).
RNA extraction method: Chaotropic lysis (guanidine thiocyanate) followed by RNA adsorption onto silica particles to remove RT‑PCR inhibitors.
Real-time RT‑PCR: Amplification and detection with hydrolysis probes for sensitive, specific quantification and confirmation during amplification.
Controls and calibration:
- Process control virus (e.g., mengo virus) to monitor extraction and processing.
- Linear dsDNA control for quantification standards.
- External control (EC) RNA and negative controls to assess inhibition and contamination.
Data interpretation: Construction of standard curves, calculation of extraction efficiency, assessment of RT‑PCR inhibition, and expression of results with defined precision and reproducibility data.
Quality requirements: Laboratory practices, equipment, reagent preparation, plate layouts and validation guidance (see annexes).

Applications and users

ISO 15216-1:2017 is intended for:

Food testing laboratories performing viral contamination testing of soft fruit, vegetables, bottled water, BMS and food-contact surfaces.
Public health agencies and food safety authorities needing standardized quantitative viral RNA data for outbreak investigation and surveillance.
Industry quality teams and third-party testing services requiring harmonized methods for HAV and norovirus monitoring and risk assessment.

Related standards

ISO 15216 series (other parts and validation guidance)
ISO 22119, ISO 22174, ISO 20838 (PCR/real-time PCR and microbiology of food) For authoritative details and annex content (primers, mastermixes, validation data), consult the full ISO 15216-1:2017 document on the ISO website.

ISO 15216-1:2017 PDF

Technical details

Abstract

Overview

Key topics and technical requirements

Applications and users

Related standards

Related standards

Abstract

Overview

Key topics and technical requirements

Applications and users

Related standards

Related standards

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