Detection of Infectious Hepatitis E Virus in Retail Pork Pâté After Optimization of Extraction Protocols

Hepatitis E virus (HEV) is an emerging foodborne pathogen in Europe, primarily associated with the consumption of pork products. While conventional destructive TRIzol extraction methods followed by RT-qPCR can detect HEV RNA in processed meat, this approach fails to distinguish between infectious and inactivated virus, complicating risk assessment. Currently, no standardized method exists for extracting intact, infectious HEV from pork products, limiting our ability to evaluate viral infectivity post-processing. In this study, we developed and evaluated three extraction methods to recover intact, infectious HEV from pork meat products. Our optimized soft extraction method using TGBE buffer and stomacher-based homogenization allowed us to successfully isolate infectious HEV from pork liver pâté bought in a Belgian supermarket, confirming the presence of infectious HEV in a ready-to-eat pâté intended for direct consumer consumption. Furthermore, viral RNA was detected following extraction with this method at concentrations (copies/g) similar to those obtained with the TRIzol method, while preserving viral integrity, as confirmed by capsid integrity assays and cell culture infectivity models. As a critical tool for assessing the impact of food processing on viral infectivity, the method holds significant potential for advancing food safety research and improving risk evaluations related to HEV transmission through pork products.