Growth of infectious salmon anaemia virus in CHSE-214 cells and evidence for phenotypic differences between virus strains.
Infectious salmon anaemia virus (ISAV) is a new orthomyxovirus-like virus. Thirteen isolates of ISAV (11 from Canada, one from Norway and one from Scotland) were studied for their replication in the CHSE-214 cell line compared with that in the SHK-1 cell line. All isolates replicated in SHK-1 cells, producing CPE between 3 and 12 days post-inoculation (p.i.). Six Canadian isolates also replicated in CHSE-214 cells, with production of CPE between 4 and 17 days p.i. Analysis of a one-step growth curve of ISAV in CHSE-214 cells showed that progeny virions remained predominantly cell-associated, accounting for the focalized nature of the CPE in the cell monolayer. One isolate (HKS 36) replicated in CHSE-214 cells, as shown by positive RT-PCR results of blind passages, but was non-cytopathic. All of the isolates were analysed for genetic heterogeneity by RT-PCR and RFLP with EcoRI and XhoI in a fraction of genome segment 2. The Canadian isolates showed a different RFLP profile to those of isolates Glesvaer/2/90 from Norway and 390/98 from Scotland. Structural proteins of four isolates, 'Back Bay 98', RPC/NB-877, RPC/NB-049 and Glesvaer/2/90, were examined further by SDS-PAGE. All viruses showed four major polypeptides, designated here as VP1-VP4, in Coomassie blue-stained gels. In isolates Glesvaer/2/90 and RPC/NB-877, these viral proteins had estimated molecular masses of 74, 53, 46 and 26.5 kDa, respectively. Viral proteins in isolates 'Back Bay 98' and RPC/NB-049 were of similar sizes, except that VP3 was 43 kDa. Taken together, these results show that there are phenotypic differences among strains of ISAV.
Kibenge
F S
FS
Department of Pathology and Microbiology, AVC Inc. and Department of Health Management(3), Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PEI, Canada. kibenge@upei.ca
Lyaku
J R
JR
Rainnie
D
D
Hammell
K L
KL
D000818:Animals; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral; D005455:Fluorescent Antibody Technique; D016679:Genome, Viral; D008854:Microscopy, Electron; D009975:Orthomyxoviridae / Q000145:classification* / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D010641:Phenotype; D012150:Polymorphism, Restriction Fragment Length; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012474:Salmon / Q000821:virology*; D015678:Viral Structural Proteins / Q000235:genetics; D014779:Virus Replication
D015678:Viral Structural Proteins