Fachbereich Veterinärmedizin



    Detection of Bovine Viral Diarrhea Virus RNA in Formalin-fixed, Paraffin-embedded Brain Tissue by Nested Polymerase Chain Reaction (1993)

    Zeitschriftenartikel / wissenschaftlicher Beitrag
    Gruber, A D (WE 12)
    Greiser-Wilke, I M
    Haas, L
    Hewicker-Trautwein, M
    Moennig, V
    Journal of Virological Methods; 43(3) — S. 309–319
    ISSN: 0166-0934
    Pubmed: 8408445
    Institut für Tierpathologie

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    Gebäude 12
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    Abstract / Zusammenfassung

    Isolation and amplification of RNA from formalin-fixed, paraffin-embedded tissues is delicate due to its fragility and ubiquitous ribonucleases. For retrospective studies, however, a convenient procedure for the detection of RNA in archived material is of great value. Bovine viral diarrhea (BVD) virus is a member of the pestivirus genus in the family Flaviviridae. Different protocols for the isolation of BVD virus RNA from fresh and autolytic as well as from routinely formalin-fixed and paraffin-embedded brain tissue of BVDV-infected calves were compared. The polymerase chain reaction (PCR) after reverse transcription (RT-PCR) was carried out subsequently for the detection of viral RNA. Using proteinase K digestion of deparaffinized tissue sections without additional ribonuclease inhibitors and subsequent nested PCR, a 803 bp fragment of the gene coding for the nonstructural protein p125 of BVD virus could be consistently detected. In addition, BVD virus RNA was detected by RT-PCR from non-fixed brain tissue after 10 days of autolysis.