Validation of EN ISO method 15216 - Part 1 – Quantification of hepatitis A virus and norovirus in food matrices

Abstract

Hepatitis A virus (HAV) and norovirus are important agents of food-borne human viral illness, with common vehicles including bivalve molluscan shellfish, soft fruit and various vegetables. Outbreaks of viral illness due to contamination of the surfaces of foods, or food preparation surfaces by for example infected food handlers are also common. Virus analysis of food matrices can contribute towards risk management for these hazards and a two-part technical specification for determination of Hepatitis A virus and norovirus in food matrices (ISO/TS 15216:2013) was published jointly by the European Committee for Standardisation and the International Organization for Standardization in 2013. As part of the European Mandate No. M381 to validate 15 standards in the field of food microbiology, an international validation study involving 18 laboratories from 11 countries in Europe was conducted between 2012 and 2014. This study aimed to generate method characteristics including limit of detection, limit of quantification, repeatability and reproducibility for ISO 15216 - Part 1, the method for quantification, in seven food matrices. The organization and results of this study, including observations that led to improvements in the standard method are presented here. After its conclusion, the method characteristics generated were added to the revised international standard, ISO 15216-1:2017, published in March 2017.

Effectiveness of Consumers Washing with Sanitizers to Reduce Human Norovirus on Mixed Salad

“…Virus extraction was performed following the ISO 15216-1:2017 [23]. Limit of detection of the analysis of HuNoV on lettuce using the ISO 15216-1:2017 was of 0.46 and 0.88 copies/g for GI and GII, respectively [24]. Briefly, 25 g of sample was added to 40 mL of Tris/glycine/beef extract (TGBE) buffer pH 9.5 (100 mM Tris-HCl, 50 mM glycine, 1% beef extract) and 10 µL of Mengovirus as a process control virus material.…”

Glass Wool Concentration Optimization for the Detection of Enveloped and Non-enveloped Waterborne Viruses

“…The choice of PEG precipitation over other possible reconcentration options was also based on the affordability of the procedure in order to keep the cost of the entire virus concentration, i.e., glass wool adsorption/elution and PEG reconcentration, low enough (below 15 euro per sample) to be applicable for environmental surveillance studies performed in developing areas. Current ISO standards for the molecular detection of viruses in water and food specify a threshold of acceptance of virus recovering efficiencies above 1% (International Organization for Standardization -ISO 2017; Lowther et al 2017). This criterion is met for the recovery of nonenveloped and enveloped viruses from large volumes (50 L) of water employing glass wool filtration and PEG precipitation: 4.5% and 5.1%, respectively.…”

Hepatitis A infections from food

“…Very recently, the ISO method 15216-1 has been validated in an international study involving 18 laboratories from 11 European countries. Moreover, this study provided useful analytical parameters such as the limit of detection (LoD), the limit of quantification (LoQ), repeatability and reproducibility (Lowther et al 2019).…”

“…According to ISO 15216, HAV in bivalve molluscan shellfish is extracted from the digestive glands using a treatment with a proteinase K solution. In a validation study using Pacific oysters, this procedure resulted in a LoD and a LoQ of 198 gc g À1 (Lowther et al 2019).…”