Plaque assay and replication of Tipula iridescent virus in Spodoptera frugiperda ovarian cells.
A plaque assay was developed for the study of Tipula iridescent virus (TIV) replication using a cell line derived from the fall army worm Spodoptera frugiperda (Sf9). Infection and plaque formation were monitored with time by phase contrast microscopy, video and fluorescent light microscopy. Structure of virions, viroplasmic centres and organelles of infected cells were examined by transmission electron microscopy (TEM). After 4 h postinfection, plaques were visibly detected within the cell monolayer by the presence of localized cell damage and production of numerous vesicular-like cytoplasmic structures. Quantitation of virions present per A260 unit of TIV preparation was determined by TEM. The number of visible plaques corresponded to virus concentration and 1 A260 produced approximately 10(5) plaques. DNA hybridization analysis revealed no gross differences in genomic DNA from TIV propagated in either Sf9 cells or wax moth Galleria mellonella larvae. These findings indicate that Sf9 is permissive for replication of TIV and superior by some parameters to other cell lines currently in use for the study of host cell/TIV interactions.
Institute of Biological Sciences, National Research Council, Ottawa, Ontario.
M
M
Czuba
S
S
Tajbakhsh
T
T
Walker
M J
MJ
Dove
B F
BF
Johnson
V L
VL
Seligy
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003588:Cytopathogenic Effect, Viral; D005260:Female; D007304:Insect Viruses / Q000254:growth & development*; D007497:Iridoviridae / Q000254:growth & development*; D008969:Molecular Sequence Data; D009036:Moths / Q000821:virology; D010053:Ovary / Q000166:cytology / Q000648:ultrastructure / Q000821:virology*; D018411:Spodoptera / Q000166:cytology / Q000648:ultrastructure / Q000821:virology*; D010948:Viral Plaque Assay / Q000379:methods*