Production of Plaques in Monolayer Tissue Cultures by Single Particles of an Animal Virus

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Abstract
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.

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