SIST EN ISO 15216-1:2017

Overview

EN ISO 15216-1:2017 (CEN / ISO 15216-1:2017) specifies a horizontal method for quantifying hepatitis A virus (HAV) and norovirus (genogroup I and II) RNA in selected food matrices and food-contact surfaces using real-time RT‑PCR. The standard covers sample processing (virus liberation), RNA extraction (lysis with guanidine thiocyanate and adsorption on silica), and amplification/detection by real-time RT‑PCR to produce quantitative results. The method is validated for specific sample types (soft fruit; leaf, stem and bulb vegetables; bottled water; bivalve molluscan shellfish; and food surfaces) and is not validated for other foodstuffs or matrices.

Key topics and technical requirements

• Scope and limitations: Quantification of HAV and norovirus GI/GII RNA in defined food matrices; not validated for multi-component foods or other viruses.
• Sample preparation: Procedures for virus liberation from foods and surfaces prior to extraction.
• RNA extraction: Lysis with guanidine thiocyanate and adsorption onto silica-based matrices.
• Real-time RT‑PCR: Amplification and detection of target sequences; generation of standard curves for quantification.
• Controls and quality checks:
  • Process control virus (e.g., mengo virus) to monitor extraction recovery.
  • Double-stranded DNA (dsDNA) controls and external calibration (EC RNA) for assay performance.
  • Negative process controls to detect contamination.
• Data interpretation: Calculation of extraction efficiency, RT‑PCR inhibition, sample quantification, and reporting units.
• Performance and validation: Includes precision data (repeatability, reproducibility) and interlaboratory study outcomes.
• Supportive annexes: Primer/probe sequences, mastermix and cycling recommendations, methods for generating controls and example plate layouts.

Applications and who uses it

EN ISO 15216-1:2017 is used by:

• Food microbiology and public health laboratories for quantitative testing of HAV and norovirus in high-risk foods.
• Regulatory agencies and food safety authorities for surveillance and outbreak investigation.
• Contract testing laboratories serving produce growers, shellfish producers, and bottlers.
• Research teams validating interventions to reduce viral contamination on foods and surfaces.

Practical implementation supports quality-controlled virus quantification, helps assess consumer risk, verifies process controls, and supports regulatory compliance for selected food categories.

Related standards

• Part of the ISO 15216 series and horizontal methods for virus detection in the food chain. This edition supersedes earlier technical specifications (CEN ISO/TS 15216-1:2013) and aligns with CEN/TC 275 and ISO/TC 34 guidance.

Keywords: ISO 15216-1:2017, hepatitis A virus, norovirus, real-time RT‑PCR, food microbiology, virus quantification, RNA extraction, guanidine thiocyanate, silica adsorption, process control.