f Detection of phocid distemper virus RNA in seal tissues using slot hybridization and the polymerase chain reaction amplification assay: genetic evidence that the virus is distinct from canine distemper virus
- Authors: Ludwig Haas†, Shaila M. Subbarao‡>, Timm Harder, Bernd Liess, Thomas Barrett
- J. Gen. Virol., April 1991 72: 825-832, doi: 10.1099/0022-1317-72-4-825
- Subject: Animal
- Published Online:
Detection of phocid distemper virus RNA in seal tissues using slot hybridization and the polymerase chain reaction amplification assay: genetic evidence that the virus is distinct from canine distemper virus, Page 1 of 1< Previous page | Next page > /docserver/preview/fulltext/jgv/72/4/JV0720040825-1.gif
Slot hybridization and the polymerase chain reaction (PCR) after reverse transcription (RT) were used to detect RNA extracted from tissues of seals after naturally occurring disease and experimental infection with phocid distemper virus (PDV). A phosphoprotein (P) gene-specific cDNA served as a probe for both slot hybridization and the identification of PCR-generated fragments by Southern blotting. As primers for the PCR assay PDV P gene-derived oligonucleotides were used. Hybridization, PCR and partial nucleic acid sequence analysis clearly demonstrated that PDV is a distinct virus (most closely related to canine distemper virus) within the morbillivirus group. PCR, when combined with Southern blot hybridization, was clearly superior to slot hybridization and more sensitive than cell culture isolation and immunofluorescence assays for the detection of virus in tissues. Considerable amounts of viral RNA could be demonstrated in the lungs and spleens. In experimentally infected animals a large quantity of virus-specific RNA was additionally found in colon samples. Using RT-PCR in combination with Southern blotting, PDV could be demonstrated in buffy coat cells using a simple and fast cell lysis procedure.
Present address: New York State College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, U.S.A.
Permanent address: Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560012, India.
© Society for General Microbiology 1991 | Published by the Microbiology Society
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