Establishment of a serum-free culture cell line, CPK-NS, which is useful for assays of classical swine fever virus.

A stable porcine kidney cell line, CPK-NS, was established and maintained in serum-free culture. A cytopathic effect (CPE) was observed clearly in CPK-NS cells infected with some classical swine fever virus (CSFV) strains which did not show the exaltation of Newcastle disease virus (END) phenomenon. Chromosome condensation and DNA fragmentation, a marker for apoptosis, were detected in cells infected with END phenomenon-negative CSFV strains. By using the CPE induced by infection with an END phenomenon-negative CSFV strain in CPK-NS cells, assays of CSFV were established. The virus titer determined in CPK-NS cells shows a high correlation with the usual peroxidase-linked assay, dome disappearance method and END method. Furthermore, the antibody titer by neutralizing test with CPK-NS cells also correlated with that measured by the usual neutralizing peroxidase-linked assay and dome disappearance method. These stable CPK-NS cells have the great advantage that a clear CPE was caused by infection with END phenomenon-negative CSFV strains and bovine serum is not necessary for cell culture and virus assays.