Comparison of Different In Situ Hybridization Techniques for the Detection of Various RNA and DNA Viruses
Vanessa M. Pfankuche,
Kerstin Hahn,
Rogier Bodewes,
Florian Hansmann,
André Habierski,
Ann-Kathrin Haverkamp,
Stephanie Pfaender,
Stephanie Walter,
Christine Baechlein,
Alexander Postel,
Eike Steinmann,
Paul Becher,
Albert Osterhaus,
Wolfgang Baumgärtner,
Christina Puff
Affiliations
Vanessa M. Pfankuche
Department of Pathology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
Kerstin Hahn
Department of Pathology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
Rogier Bodewes
Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, 3584 Utrecht, The Netherlands
Florian Hansmann
Department of Pathology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
André Habierski
Department of Pathology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
Ann-Kathrin Haverkamp
Department of Pathology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
Stephanie Pfaender
Institute for Experimental Virology, Twincore Centre for Experimental and Clinical Infection Research, Medical School Hannover (MHH)-Helmholtz Centre for Infection Research (HZI), 30625 Hannover, Germany
Stephanie Walter
Institute for Experimental Virology, Twincore Centre for Experimental and Clinical Infection Research, Medical School Hannover (MHH)-Helmholtz Centre for Infection Research (HZI), 30625 Hannover, Germany
Christine Baechlein
Institute of Virology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
Alexander Postel
Institute of Virology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
Eike Steinmann
Institute for Experimental Virology, Twincore Centre for Experimental and Clinical Infection Research, Medical School Hannover (MHH)-Helmholtz Centre for Infection Research (HZI), 30625 Hannover, Germany
Paul Becher
Institute of Virology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
Albert Osterhaus
Center for Systems Neuroscience, 30559 Hannover, Germany
Wolfgang Baumgärtner
Department of Pathology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
Christina Puff
Department of Pathology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
In situ hybridization (ISH) is a technique to determine potential correlations between viruses and lesions. The aim of the study was to compare ISH techniques for the detection of various viruses in different tissues. Tested RNA viruses include atypical porcine pestivirus (APPV) in the cerebellum of pigs, equine and bovine hepacivirus (EqHV, BovHepV) in the liver of horses and cattle, respectively, and Schmallenberg virus (SBV) in the cerebrum of goats. Examined DNA viruses comprise canine bocavirus 2 (CBoV-2) in the intestine of dogs, porcine bocavirus (PBoV) in the spinal cord of pigs and porcine circovirus 2 (PCV-2) in cerebrum, lymph node, and lung of pigs. ISH with self-designed digoxigenin-labelled RNA probes revealed a positive signal for SBV, CBoV-2, and PCV-2, whereas it was lacking for APPV, BovHepV, EqHV, and PBoV. Commercially produced digoxigenin-labelled DNA probes detected CBoV-2 and PCV-2, but failed to detect PBoV. ISH with a commercially available fluorescent ISH (FISH)-RNA probe mix identified nucleic acids of all tested viruses. The detection rate and the cell-associated positive area using the FISH-RNA probe mix was highest compared to the results using other probes and protocols, representing a major benefit of this method. Nevertheless, there are differences in costs and procedure time.
