Development of an Extraction Method to Detect Hepatitis A Virus, Hepatitis E Virus, and Noroviruses in Fish Products

Viruses are a leading cause of foodborne disease worldwide. Hepatitis viruses (hepatitis A (HAV) and hepatitis E (HEV)) and human norovirus are recognized as the main viruses of public health concern in food hygiene. ISO 15216 approved procedures are not validated for detection of HAV and human norovirus in foodstuffs, such as fishes, leading to an inability to ensure the safety of these products. This study aimed to provide a rapid and sensitive method for detecting these targets in fish products. An existing method that includes proteinase K treatment was selected for further validation using artificially contaminated fish products, according to the recent international standard ISO 16140-4. Recovery efficiencies in pure RNA extracts of viruses ranged from 0.2% to 66.2% for HAV, 4.0% to 100.0% for HEV, 2.2% to 100.0% for norovirus GI, and 0.2% to 12.5% for norovirus GII. LOD50 values were between 144 and 8.4 x 10(4) genome copies/g for HAV and HEV, and 10(4) and 2.0 x 10(3) copies/g for norovirus GI and norovirus GII, respectively. LOD95 values were between 3.2 x 10(3) and 3.6 x 10(5) genome copies/g for HAV and HEV, and between 8.8 x 10(3) and 4.4 x 10(4) genome copies/g for norovirus GI and norovirus GII, respectively. The method developed here was successfully validated in various fish products and can be applied for routine diagnostic needs.

Automated summary

Viruses, including hepatitis A (HAV), hepatitis E (HEV), and human norovirus, are major concerns in food hygiene. Existing ISO 15216 approved methods fail to validate the detection of HAV and human norovirus in food products, such as fish, jeopardizing product safety. This study aims to develop a rapid and sensitive method for detecting these viruses in fish. The method, validated according to ISO 16140-4, demonstrated efficient recovery and accurate LOD values, making it suitable for routine diagnostics in various fish products.