Sequence-independent VIDISCA-454 technique to discover new viruses in canine livers.
In many mammals, viruses cause hepatitis. Despite many efforts a specific virus responsible for canine idiopathic hepatitis has not been identified. The discovery of a viral etiology in canine hepatitis will promote the development of specific drugs and vaccines for the treatment of idiopathic hepatitis in dogs. The objective of this study was the application of the sequence-independent Virus Discovery cDNA-amplified fragment length polymorphism (VIDISCA) technique combined with high through-put sequencing on a Roche-454 sequencer to identify unknown viruses. Liver tissue of a dog with idiopathic acute hepatitis was cultured on a canine liver cell line and the cell culture medium was submitted to the VIDISCA-454 technique. Without prior knowledge of the viral species involved, this technique identified Canine adenovirus type 1 (CAV-1) as the infecting agent. This demonstrates the power of VIDISCA-454 to identify viruses, independent of preliminary information about the genomic sequence. Consequently, the strategy of propagation in this cell line followed by the VIDISCA-454 technique is valuable to identify the viral etiology of idiopathic hepatitis in dogs.
Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 104, Utrecht 3584 CM, The Netherlands. M.vanderHeijden@uu.nl
Mitzi
M
van der Heijden
Michel
M
de Vries
Frank G
FG
van Steenbeek
Robert P
RP
Favier
Martin
M
Deijs
Bas
B
Brinkhof
Jan
J
Rothuizen
Lia
L
van der Hoek
Louis C
LC
Penning
D006516:Adenoviruses, Canine / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D000818:Animals; D002460:Cell Line; D004285:Dogs; D006522:Hepatitis, Infectious Canine / Q000821:virology*; D008099:Liver / Q000821:virology*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods; D014773:Virology / Q000379:methods*; D014776:Virus Cultivation