Sakoda Y, Hikawa M, Tamura T, Fukusho A
Department of Exotic Diseases, National Institute of Animal Health, Kodaira, Tokyo, Japan.
J Virol Methods. 1998 Nov;75(1):59-68. doi: 10.1016/s0166-0934(98)00098-6.
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.
建立了一种稳定的猪肾细胞系CPK-NS，并在无血清培养条件下进行维持。在感染某些不表现新城疫病毒（NDV）蚀斑形成增强（END）现象的经典猪瘟病毒（CSFV）毒株的CPK-NS细胞中，可明显观察到细胞病变效应（CPE）。在感染END现象阴性CSFV毒株的细胞中检测到染色体浓缩和DNA片段化，这是细胞凋亡的一个标志。利用END现象阴性CSFV毒株感染CPK-NS细胞所诱导的CPE，建立了CSFV检测方法。在CPK-NS细胞中测定的病毒滴度与常用的过氧化物酶联检测法、蚀斑消失法和END法高度相关。此外，用CPK-NS细胞进行中和试验所得到的抗体滴度也与用常用的中和过氧化物酶联检测法和蚀斑消失法测得的结果相关。这些稳定的CPK-NS细胞具有很大的优势，即感染END现象阴性CSFV毒株可引起明显的CPE，并且细胞培养和病毒检测无需使用牛血清。
