PRODUCTION OF PLAQUES IN MONOLAYER TISSUE CULTURES BY SINGLE PARTICLES OF AN ANIMAL VIRUS
PRODUCTION OF PLAQUES IN MONOLAYER TISSUE CULTURES BY SINGLE PARTICLES OF AN ANIMAL VIRUS
复制标题
DOI：
10.1073/pnas.38.8.747
复制
发表时间：
1952-01-01
影响因子：
11.1
通讯作者：
DULBECCO, R
中科院分区：
文献类型：
--
作者：
DULBECCO, R
描述了一种从鸡胚制备细胞悬液的方法。在孵化的最初48小时内，细胞（成纤维细胞）在烧瓶底部形成了一层连续的单细胞层。该细胞层被西方马脑脊髓炎病毒感染，然后在细胞层上倒一层融化的营养琼脂。在37℃下3天后，可见直径为2 - 4毫米的圆形坏死区域（空斑），并证明是由病毒活动产生的。病毒浓度和空斑数量之间呈线性关系，表明每个空斑是由一个病毒颗粒产生的。进行了一些测试以确定在上述条件下能够产生空斑的颗粒数量与通过在绒毛尿囊膜上的标准接种程序能够感染鸡胚的颗粒数量之间的关系。比较结果表明，空斑数量和感染性病毒颗粒数量之间的比例约为1∶11。使用空斑计数技术一个烧瓶所获得的准确性与使用里德 - 明奇技术不少于100个鸡胚所获得的准确性相当。西方马脑脊髓炎病毒产生的空斑可与新城疫病毒产生的空斑区分开来。
A method is descr. for preparing a cell suspension from chicken embryos. During the first 48 hrs. of incubation the cells (fibroblasts) produced a single continuous cell layer over the bottom of the flask. The cell layer was infected with the virus of Western Equine Encephalo-myelitis and a layer of melted nutrient agar was poured over the cell layer. After 3 days at 37[degree] C round, necrotic areas (plaques) 2-4 mm. in diam. were visible and were proven to be produced by virus activity. A linear relationship between virus concn. and no. of plaques indicated that each plaque was produced by one virus particle. Tests were run to determine the relationship between the number of particles capable of producing a plaque under the conditions outlined above and the number of particles able to infect the chicken embryo by the standard inoculation procedure on the chorioallantoic membrane. The comparisons indicated an approx. 11 ratio between number of plaques and number of infective virus particles. The accuracy obtained with one flask using the plaque count technic was equal to that obtained with not less than 100 chicken embryos in the Reed and Muench technic. The plaques produced by Western Equine Encephalomyelitis virus were distinguishable from those produced by the Newcastle Disease virus.