ISO 15216-2:2019

Overview

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" specifies a standardized molecular method for detecting hepatitis A virus (HAV) and norovirus genogroups I (GI) and II (GII) in selected food and surface matrices. The method uses real‑time RT‑PCR and is validated for specific matrices only: soft fruit; leaf, stem and bulb vegetables; bottled water; bivalve molluscan shellfish (BMS); and food contact or food preparation surfaces. It is not validated for other foodstuffs, multi‑component foods or other matrices.

Key topics and technical requirements

• Matrix‑specific virus extraction methods:
  • Swabbing for surfaces
  • Elution with agitation and PEG/NaCl precipitation for soft fruit and vegetables
  • Adsorption–elution using positively charged membranes plus ultrafiltration for bottled water
  • Proteinase K digestion of digestive gland tissue for BMS
• RNA extraction: common approach employing capsid disruption with chaotropic reagents and RNA adsorption to silica particles.
• Real‑time RT‑PCR: use of hydrolysis probes for sensitive, specific detection and confirmation during amplification.
• Comprehensive control suite:
  • Process control virus (e.g., mengo virus) to monitor extraction and assay performance
  • External control RNA (EC RNA) for standard curve generation
  • Negative process and amplification controls; checks for RT‑PCR inhibition
• Performance & validation: the document includes method performance characteristics, limits of detection (LOD), and results from validation studies.
• Laboratory requirements: defined procedures, reagent preparation, reaction layouts and interpretation rules are provided (see normative and informative annexes).

Applications and who uses it

ISO 15216-2:2019 is intended for:

• Public health and clinical laboratories investigating foodborne viral outbreaks
• Food testing laboratories performing routine monitoring of high‑risk products (soft fruit, BMS, bottled water)
• Shellfish harvest managers and water quality authorities assessing viral contamination
• Food producers and processors implementing food safety testing programs
• Regulatory bodies and accredited laboratories requiring standardized, comparable methods for virus detection in food using real‑time RT‑PCR

Practical uses include outbreak source tracking, routine surveillance, supplier verification and compliance testing where HAV and norovirus are of concern.

Related standards

• ISO 15216 series (other parts)
• ISO 20838 - PCR requirements for qualitative methods
• ISO 22119 - Real‑time PCR general requirements for food pathogens
• ISO 22174 - PCR general requirements and definitions

Keywords: ISO 15216-2:2019, hepatitis A virus, norovirus, real‑time RT‑PCR, food safety testing, bivalve molluscan shellfish, bottled water, virus detection, RNA extraction, PCR inhibition.