牛冠状病毒的细胞培养增殖
Cell culture propagation of bovine coronavirus.
作者信息
Saif Linda J, Heckert Robert A, Miller Kathy L, Tarek Mohamed M
机构信息
1Food Animal Health Research Program, Ohio Agricultural Research and Development Center, Ohio State University, 44691 Wooster, Ohio, Ohio.
2Department of Parasitology and Microbiology, Faculty of Medicine, Zagazig University, Zagazig, Egypt.
出版信息
J Tissue Cult Methods. 1988;11(3):139-145. doi: 10.1007/BF01404267.
Although most field strains of bovine coronavirus (BCV) grow poorly in cell culture and fail to produce cytopathic effects (CPE) until after blind passage, primary calf kidney (PCK) and Vero cells have permitted primary isolation of virus. Cell culture-adapted strains of BCV replicate in PCK, bovine embryonic lung, bovine fetal thyroid, bovine fetal brain, bovine skin cells, ovine fetal kidney cells, and the cell lines pig kidney K3 and 15, Vero, human embryonic lung fibroblasts, HRT-18, MDBK and BEK-1, with trypsin useful for enhancing replication. Organ culture as well as suckling mouse, rat, and hamster brains also support the growth of cell culture-adapted BCV strains. Viral growth is most commonly detected by CPE, immunofluorescence, hemagglutination, and hemadsorption assays or electron microscopy of supernatants from infected cells. In this report, the optimal conditions for the growth and plaque assay of the NCDV strain of BCV in MDBK cells are described.
尽管大多数牛冠状病毒（BCV）野毒株在细胞培养中生长不佳，在盲传后才产生细胞病变效应（CPE），但原代犊牛肾（PCK）细胞和Vero细胞可用于病毒的初次分离。适应细胞培养的BCV毒株可在PCK细胞、牛胚胎肺细胞、牛胎儿甲状腺细胞、牛胎儿脑细胞、牛皮肤细胞、绵羊胎儿肾细胞以及猪肾K3和15、Vero、人胚胎肺成纤维细胞、HRT-18、MDBK和BEK-1细胞系中复制，胰蛋白酶有助于增强复制。器官培养以及乳鼠、大鼠和仓鼠的脑也支持适应细胞培养的BCV毒株生长。病毒生长最常通过CPE、免疫荧光、血凝、血吸附试验或对感染细胞上清液进行电子显微镜检测来检测。在本报告中，描述了BCV的NCDV毒株在MDBK细胞中生长和空斑试验的最佳条件。
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