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 following improved method for detecting Dengue viral RNA in tissue sections was devised. Reverse transcription of the viral RNA to DNA was followed by the in situ polymerase chain reaction to amplify the viral nucleic acid. This was followed by DNA hybridization with an alkaline phosphatase-labelled probe. The enzyme was then reacted with Fast red and counterstained with hematoxylin. Viral nucleic acid was readily demonstrated within glial cells and macrophages in brains of animals infected with two different serotypes of Dengue.
Lucia
H L
HL
Department of Pathology, University of Mississippi Medical Center, Jackson 39216.
Kangwanpong
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D000818:Animals; D000833:Animals, Suckling; D001483:Base Sequence; D001921:Brain / Q000821:virology; D015342:DNA Probes; D003716:Dengue Virus / Q000302:isolation & purification*; D005260:Female; D017403:In Situ Hybridization; D008264:Macrophages / Q000821:virology*; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D009457:Neuroglia / Q000821:virology*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling
D015342:DNA Probes; D012367:RNA, Viral