Hepatitis E virus (HEV) is considered a potentially emerging foodborne viral pathogen in Europe, with the ability of zoonotic transmission. More specifically, genotypes 3 and 4 (gt3 and gt4) have been confirmed as zoonotic genotypes. Study on detection and the zoonotic potential of HEV gt3 and gt4 has been hampered by the lack of a reliable HEV cultivation system, but recent advances have been made using a 3D cell culture. Therefore, a first objective of this project proposal involves the development of an innovative real-time RTPCR assay for detection of gt3 and gt4 HEV, including a high resolution melting analysis. This assay will be combined with methods for extraction of HEV from tissues and foods. The developed HEV detection assay will be tested during a sampling of Belgian pig farms and slaughterhouses, which will also generate data on HEV prevalence in Belgium. The second objectives emphasizes on the interactions between HEV and host cells of zoonotic (gt3 and gt4) and non-zoonotic (gt1 and gt2) HEV, making use of the recently developed 3D cell culture system. The binding efficiency and replication efficiency of HEV will be investigated using the molecular method and strains obtained during samplings described in the first objective.