Overview
EN ISO 15216-2:2019 - Microbiology of the food chain - Horizontal method for determination of hepatitis A virus (HAV) and norovirus (GI and GII) using real‑time RT‑PCR - Part 2: Method for detection - specifies a standardized laboratory method for detecting these viral pathogens in specified foods and surfaces. The method covers virus extraction, RNA purification and real‑time RT‑PCR detection and includes controls and interpretation criteria. It applies to soft fruit, leaf/stem/bulb vegetables, bottled water, bivalve molluscan shellfish (BMS) and surfaces. The standard is not validated for other food matrices (including multi‑component foods) or for detection of other viruses.
Key topics and technical requirements
- Target viruses: Hepatitis A virus (HAV), Norovirus genogroups I (GI) and II (GII).
- Method technology: Real‑time reverse transcription PCR (real‑time RT‑PCR) for sensitive molecular detection.
- Scope limitations: Validated only for the listed foodstuffs and surfaces; not for other matrices or viruses.
- Sample processing: Defined procedures for virus extraction from each matrix (soft fruit, vegetables, BMS, bottled water, surfaces).
- RNA extraction and controls: Includes requirements for RNA extraction, use of a process control virus (e.g., mengovirus) and external control RNA (EC RNA) to monitor extraction efficiency and RT‑PCR performance.
- Quality controls and inhibition checks: Procedures to assess RT‑PCR inhibition, construct standard curves for controls, and calculate extraction efficiency.
- Performance and validation: Contains guidance on method validation, limits of detection (LOD), specificity, sensitivity and test reporting.
- Documentation: Specifies required test report elements and typical plate/layouts (annexes provide primers/probes, mastermixes, reagent recipes and validation data).
Practical applications
- Routine surveillance and outbreak investigation for foodborne viral contamination in declared food matrices (e.g., retail soft fruit, shellfish monitoring, bottled water quality).
- Verification testing by food safety laboratories, shellfish harvesters and water testing labs to detect HAV and norovirus contamination.
- Support for regulatory compliance, risk assessment and public health response when viral contamination of food or contact surfaces is suspected.
Who should use this standard
- Public health and food microbiology laboratories
- Commercial and regulatory food testing laboratories
- Shellfish producers and water quality testing facilities
- Food safety consultants and auditors involved in viral hazard control
Related standards
- Consult other parts of the ISO 15216 series and national guidance for quantification, broader validation and laboratory accreditation requirements (e.g., complementary documents covering quantification and laboratory quality systems).
Keywords: EN ISO 15216-2:2019, hepatitis A virus, norovirus GI GII, real-time RT-PCR, food testing, bivalve molluscan shellfish, bottled water, soft fruit, food microbiology, virus detection.
