Evidence for Novel Hepaciviruses in Rodents

Drexler, Jan Felix; Corman, Victor Max; Müller, Marcel Alexander; Lukashev, Alexander N.; Gmyl, Anatoly; Coutard, Bruno; Adam, Alexander; Ritz, Daniel; Leijten, Lonneke M.; van Riel, Debby; Kallies, Rene; Klose, Stefan M.; Gloza-Rausch, Florian; Binger, Tabea; Annan, Augustina; Adu-Sarkodie, Yaw; Oppong, Samuel; Bourgarel, Mathieu; Rupp, Daniel; Hoffmann, Bernd; Schlegel, Mathias; Kümmerer, Beate M.; Krüger, Detlev H.; Schmidt-Chanasit, Jonas; Setién, Alvaro Aguilar; Cottontail, Veronika M.; Hemachudha, Thiravat; Wacharapluesadee, Supaporn; Osterrieder, Klaus; Bartenschlager, Ralf; Matthee, Sonja; Beer, Martin; Kuiken, Thijs; Reusken, Chantal; Leroy, Eric M.; Ulrich, Rainer G.; Drosten, Christian

HCV immunofluorescence assay. An indirect immunofluorescence assay (IFA) was done using HCV-infected HuH7-cells (strain JC1) or replicon JFH1-transfected cells. Cells were fixed with paraformaldehyde (4%), permeabilized with 0.5% Triton X-100 in 16PBS for 5 minutes and processed as described previously [46] . Bat sera were diluted 1:50. Reactions were detected with goat-anti-bat Ig (Bethyl, 1:1000) and cyanine 2 (Cy2)-labelled donkey-anti-goat Ig (Dianova, 1:100). For control reactions, a polyclonal rabbit NS3-Ig raised against the NS3 helicase domain of JFH1 (1:400) and an Alexa 568-conjugated goat-anti-rabbit Ig (Invitrogen, 1:1000) were used.