[Detection of distemper virus N protein RNA in the brain of dogs with spontaneous distemper encephalitis using a digoxigenin-labeled, double-stranded DNA probe for in situ hybridization].
A digoxigenin-labelled dsDNA-probe of 287 basepairs length complementary to the nucleoprotein-gene of canine distemper virus (CDV) was generated by the polymerase-chain-reaction. The dsDNA-probe hybridized specifically with base sequences of 8 different CDV strains, whereas no hybridization was observed with a porpoise and a canine parainfluenza virus and only a weak signal was obtained with measles virus. In formalin-fixed, paraffin-embedded brain sections of 35 immunohistologically CDV antigen positive dogs with spontaneous distemper encephalitis CDV-RNA could be detected in 25 cases by in situ hybridization. The reason for the lack of RNA detection in some immunohistologically positive dogs may be due to the low stability of DNA-RNA-hybrids. Degradation of RNA by RNAses or diffusion out of autolysed cells can not be excluded.
Gaedke
K
K
Institut für Veterinär-Pathologie, Justus-Liebig-Universität in Giessen.
Teifke
J P
JP
Hardt
M
M
Alldinger
S
S
Baumgärtner
W
W
D000818:Animals; D001921:Brain / Q000821:virology*; D002213:Capsid / Q000235:genetics*; D002522:Chlorocebus aethiops; D015342:DNA Probes; D004076:Digoxigenin; D004216:Distemper / Q000175:diagnosis*; D004217:Distemper Virus, Canine / Q000235:genetics / Q000302:isolation & purification*; D004285:Dogs; D017403:In Situ Hybridization / Q000662:veterinary; D012367:RNA, Viral / Q000032:analysis*; D014709:Vero Cells; D014758:Viral Core Proteins / Q000235:genetics*
D015342:DNA Probes; D012367:RNA, Viral; D014758:Viral Core Proteins; D004076:Digoxigenin
Nachweis von Staupevirus-N-Protein-RNS im Gehirn von Hunden mit spontaner Staupeenzephalitis mittels einer Digoxigenin-markierten, doppelsträngigen DNS-Sonde in der in situ Hybridisierung.