Journal of Clinical Virology 52 (2011) 50–54 Contents lists available at ScienceDirect Journal of Clinical Virology jo u r n al hom epage: www.elsevier.com/locate/jcv Detection of hepatitis E virus (HEV) from porcine livers in Southeastern Germany and high sequence homology to human HEV isolates  Jürgen J. Wenzel ∗ , Julia Preiß, Mathias Schemmerer, Barbara Huber, Annelie Plentz, Wolfgang Jilg Institute of Medical Microbiology and Hygiene, University of Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg, Germany a r t i c l e i n f o Article history: Received 10 April 2011 Received in revised form 6 May 2011 Accepted 13 June 2011 Keywords: Hepatitis E virus Pig liver Zoonotic infection Real-time PCR Phylogenetic analysis a b s t r a c t Background: Hepatitis E virus (HEV) has been identified as an emerging cause of infectious hepatitis over the last years in developed countries. In contrast to travel associated hepatitis E, zoonotic sources of infection are suspected for autochthonous cases in Europe. Objective: Since pigs are known reservoirs of HEV, we tested porcine livers sold as food in Southeastern Germany for the presence of hepatitis E virus RNA. Study design: We purchased 200 porcine liver samples in 81 butcher shops and grocery stores in Regens- burg, Germany. Nucleic acid preparations were tested for the presence of HEV RNA by quantitative real-time PCR (RT-qPCR). HEV isolates from positive samples were characterized by partial sequencing of ORF1 and ORF2 regions in the HEV genome and by phylogenetic analysis. Results: Specimens from eight (4%) of 200 purchased pig livers had detectable HEV RNA amounts. Sequence determination and phylogenetic analysis allowed two novel isolates to be classified as HEV genotype 3, subgenotype 3a (swR437) and 3c (swR269), respectively. Both novel swine HEV isolates showed high sequence homology to isolates obtained from patients with acute HEV infection from the same geographic region. Conclusions: These results support the suggested role of undercooked pig products in food as a source of zoonotic HEV infection for humans. It remains to be clarified if this mechanism of transmission is respon- sible for the surprisingly high anti-HEV IgG prevalence recently observed in some European countries and the USA. © 2011 Elsevier B.V. All rights reserved. 1. Background Hepatitis E virus (HEV) is a small, non-enveloped virus with an icosahedral capsid of ∼33 nm in diameter. It contains a positive- sense, single-stranded RNA genome of approximately 7.2 kb in size. The genome is composed of a short 5 ′ untranslated region (UTR), three open reading frames (ORF1–3) and a short 3 ′ UTR terminated by a poly(A) tail. 1 Sequencing and analysis of several full-length viral genomes led to the identification of four major mammalian HEV genotypes (1–4) with unique geographic distribution patterns: Genotype 1 is found predominantly in Asia and Africa, genotype 2 is a Mexican strain, genotype 3 is found in industrialized coun- tries and genotype 4 strains are from sporadic cases in Asia. 2–4 HEV  The nucleotide sequences of two swine and six human HEV isolates reported herein have been assigned DDBJ/EMBL/GenBank accession nos. FR846453, FR846454, FR846455, FN995000, FN995001, FR846450, FN985025, FR846451, FR846452, FR728255, FR728256, GU479457, GU479458. ∗ Corresponding author. Tel.: +49 941 944 6411; fax: +49 941 944 6402. E-mail address: juergen.wenzel@klinik.uni-regensburg.de (J.J. Wenzel). is currently the only known member of genus Hepevirus, family Hepeviridae. 5 The virus was first identified in 1980 as the main causative agent of epidemic and endemic enterically transmitted non-A, non-B hepatitis in India. 6–8 It is now clear that genotype 1 HEV is a major cause of infectious hepatitis in many develop- ing countries in Asia, Africa and the Middle East. 4 The virus is transmitted primarily by the fecal–oral route. Water-borne epidemics are typical for areas where sanitation conditions are poor. In these regions, hepatitis E is an important public health concern. However, over the last years, studies have documented that hepatitis E infection also occurs among individuals in industri- alized countries with no recent history of travel to previously known HEV endemic countries. 9–11 In contrast to travel asso- ciated hepatitis E, zoonotic sources of HEV (genotypes 3 and 4) infection are suspected for autochthonous cases in indus- trialized countries. 12 In Germany, HEV strains of genotypes 1, 3 and 4 have been isolated from patients with acute hepati- tis E. 13–16 In a recent case control study on HEV infection in Germany, consumption of offal (including porcine liver) and wild boar meat was independently associated with autochthonous HEV 1386-6532/$ – see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.jcv.2011.06.006