Comparison of Different In Situ Hybridization Techniques for the Detection of Various RNA and DNA Viruses
AbstractIn 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
- Supplementary File 1:
ZIP-Document (ZIP, 52635 KB)
Share & Cite This Article
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.
Pfankuche VM, 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(7):384.Chicago/Turabian Style
Pfankuche, Vanessa M.; Hahn, Kerstin; Bodewes, Rogier; Hansmann, Florian; Habierski, André; Haverkamp, Ann-Kathrin; Pfaender, Stephanie; Walter, Stephanie; Baechlein, Christine; Postel, Alexander; Steinmann, Eike; Becher, Paul; Osterhaus, Albert; Baumgärtner, Wolfgang; Puff, Christina. 2018. "Comparison of Different In Situ Hybridization Techniques for the Detection of Various RNA and DNA Viruses." Viruses 10, no. 7: 384.
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.