A new assay for classical swine fever virus based on cytopathogenicity in porcine kidney cell line FS-L3.
A new assay termed the dome disappearance method for classical swine fever virus (CSFV) using FS-L3 cells with serum-free culture medium was developed. The CSFV live vaccine GPE- strain grows well and shows a slight cytopathic effect (CPE) in FS-L3 cells. This CPE results in the disappearance of the unique fluid-filled multicellular domes on a single monolayer of FS-L3 cells. By using this phenomenon, dome disappearance, as a marker of infection, it was possible to determine the titers of CSFV and its neutralizing antibody. The virus titer determined by this method shows a good correlation with that determined by immunochemical and interference methods. Furthermore, the amount of neutralizing antibody measured by this method also correlated with that measured by the Exaltation of Newcastle Disease Virus (END) neutralizing method. The dome disappearance method developed in this experiment is a simple and safe procedure and has the great advantage that bovine serum, which may contain antibody against bovine viral diarrhea virus, is not necessary for the cultivation of FS-L3 cells.
Sakoda
Y
Y
Department of Exotic Diseases, National Institute of Animal Health, Kodaira, Tokyo, Japan. sakoda@ss.ed.affrc.go.jp
Yamaguchi
O
O
Fukusho
A
A
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000097:blood; D015152:Blotting, Northern; D002460:Cell Line; D006691:Classical Swine Fever / Q000276:immunology / Q000821:virology*; D006692:Classical Swine Fever Virus / Q000276:immunology / Q000302:isolation & purification / Q000472:pathogenicity / Q000502:physiology*; D003588:Cytopathogenic Effect, Viral; D004587:Electrophoresis, Agar Gel; D019084:Fluorescent Antibody Technique, Indirect; D009500:Neutralization Tests; D013552:Swine; D014779:Virus Replication
D000914:Antibodies, Viral