ISO 15216-1:2017

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.