Novel Hepatitis E Virus Genotype in Norway Rats, Germany
Reimar Johne, Gerald Heckel, Anita Plenge-Bönig, Eveline Kindler, Christina Maresch, Jochen Reetz, Anika Schielke, and Rainer G. Ulrich
Author affiliations: Author affiliations: Federal Institute for Risk Assessment, Berlin, Germany (R. Johne, J. Reetz, A. Schielke); University of Bern, Bern, Switzerland (G. Heckel, E. Kindler); Swiss Institute of Bioinformatics, Lausanne, Switzerland (G. Heckel); Institute of Hygiene and Environment Hamburg, Hamburg, Germany (A. Plenge-Bönig); Friedrich-Loeffler-Institut, Greifswald–Insel Riems, Germany (C. Maresch, R.G. Ulrich); Free University of Berlin, Berlin (A. Schielke)
Figure 2. Immunohistochemical staining (peroxidase-antiperoxidase (PAP) technique) of liver samples from 2 rat-hepatitis E virus (HEV)–positive Norway rats from Germany, July 2009. Arrows indicate immunohistochemical positive reactions in the cytoplasm of single hepatocytes (A) and in a few foci in hepatocytes and stellate cells (B). For PAP staining, deparaffinized slides of liver samples were incubated with anti-HEV–positive human serum, which had been previously used to detect rat HEV by using solid phase immunoelectron microscopy (10), for 1 h at 37°C with protein A (Sigma-Aldrich, Steinheim, Germany) at a dilution of 1:100 for 45 min at 37°C and finally with PAP complexes from rabbits (Sigma, St. Louis, MO, USA) at a dilution 1:200 for 45 min at 37°C. AEC (3-amino-9-ethylcarbazol; Sigma Chemie GmbH, Deisenhofen, Germany) was used as the substrate chromogene. The slides were counterstained with hematoxylin and subsequently analyzed by light microscopy. Scale bars = 20 µm.
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