Enhancement of in situ hybridization to detect bovine herpesvirus 4 in paraffin sections.
In situ hybridization (ISH) protocols including different pre-treatment regimes were developed and compared for their effects on detecting bovine herpesvirus 4 (BoHV-4) in formalin fixed, paraffin embedded tissue. Results were compared for the hybridization and background signal intensities, and cellular morphology. We found that optimum results were obtained using enzyme treatment-thermal cycling as a pre-treatment of ISH. The results showed that the combination of protease and thermal cycling would be recommended as a means of supplementing in situ hybridization methods, especially when using long-term formalin fixed, paraffin-embedded tissue.
Kim
O
O
Department of Laboratory Animal Sciences, College of Medicine, Seoul National University, South Korea.
Kwak
D
D
D000818:Animals; D002417:Cattle; D002460:Cell Line; D026021:Herpesvirus 4, Bovine / Q000235:genetics / Q000302:isolation & purification*; D017403:In Situ Hybridization; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction