Sakoda Y, Yamaguchi O, Fukusho A
Department of Exotic Diseases, National Institute of Animal Health, Kodaira, Tokyo, Japan.
J Virol Methods. 1998 Jan;70(1):93-101. doi: 10.1016/s0166-0934(97)00175-4.
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.
开发了一种新的检测方法，即使用无血清培养基的FS-L3细胞检测经典猪瘟病毒（CSFV）的穹顶消失法。CSFV活疫苗GPE-株在FS-L3细胞中生长良好，并表现出轻微的细胞病变效应（CPE）。这种CPE导致FS-L3细胞单层上独特的充满液体的多细胞穹顶消失。通过利用这种穹顶消失现象作为感染的标志物，可以确定CSFV及其中和抗体的滴度。用该方法测定的病毒滴度与用免疫化学和干扰方法测定的结果具有良好的相关性。此外，用该方法测定的中和抗体量也与用新城疫病毒强化（END）中和法测定的结果相关。本实验开发的穹顶消失法是一种简单安全的方法，具有很大的优势，即培养FS-L3细胞时无需使用可能含有抗牛病毒性腹泻病毒抗体的牛血清。
