IDENTIFICATION OF DENGUE VIRUS-INFECTED CELLS IN PARAFFIN-EMBEDDED TISSUE USING IN-SITU POLYMERASE CHAIN-REACTION AND DNA HYBRIDIZATION

Using tissue from suckling mice infected with Dengue virus, the follow ing improved method for detecting Dengue viral RNA in tissue sections was devised. Reverse transcription of the viral RNA to DNA was followe d by the in situ polymerase chain reaction to amplify the viral nuclei c acid. This was followed by DNA hybridization with an alkaline phosph atase-labelled probe. The enzyme was then reacted with Fast red and co unterstained with hematoxylin. Viral nucleic acid was readily demonstr ated within glial cells and macrophages in brains of animals infected with two different serotypes of Dengue.