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Viruses 2018, 10(7), 384; https://doi.org/10.3390/v10070384

Comparison of Different In Situ Hybridization Techniques for the Detection of Various RNA and DNA Viruses

1
Department of Pathology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
2
Center for Systems Neuroscience, 30559 Hannover, Germany
3
Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, 3584 Utrecht, The Netherlands
4
Department of Viroscience, The Erasmus University Medical Center, 3015 Rotterdam, The Netherlands
5
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
6
Institute of Virology, University of Veterinary Medicine Hannover, 30559 Hannover, Germany
7
German Center for Infection Research, Partner Site Hannover-Braunschweig, 30559 Hannover, Germany
8
Department of Molecular and Medical Virology, Ruhr-University, 44780 Bochum, Germany
9
Research Center for Emerging Infections and Zoonoses (RIZ), University of Veterinary Medicine Hannover, 30559 Hannover, Germany
10
Artemis One Health, 2629 Delft, The Netherlands
*
Author to whom correspondence should be addressed.
Received: 25 May 2018 / Revised: 13 July 2018 / Accepted: 18 July 2018 / Published: 20 July 2018
(This article belongs to the Section Animal Viruses)
Full-Text   |   PDF [1560 KB, uploaded 20 July 2018]   |  

Abstract

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. View Full-Text
Keywords: chromogenic in situ hybridization; digoxigenin; DNA virus; fast red; fluorescent in situ hybridization; RNA virus; virus discovery chromogenic in situ hybridization; digoxigenin; DNA virus; fast red; fluorescent in situ hybridization; RNA virus; virus discovery
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Pfankuche, V.M.; Hahn, K.; Bodewes, R.; Hansmann, F.; Habierski, A.; Haverkamp, A.-K.; Pfaender, S.; Walter, S.; Baechlein, C.; Postel, A.; Steinmann, E.; Becher, P.; Osterhaus, A.; Baumgärtner, W.; Puff, C. Comparison of Different In Situ Hybridization Techniques for the Detection of Various RNA and DNA Viruses. Viruses 2018, 10, 384.

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