Dolan T M, Fenters J D, Fordyce P A, Holper J C
Appl Microbiol. 1968 Sep;16(9):1331-6. doi: 10.1128/am.16.9.1331-1336.1968.
A sensitive, reliable plaque assay system is described for five rhinoviruses using freshly prepared methylcellulose overlay and human embryonic diploid cells. Circular plaques with irregular edges, 2 mm in size, were formed by rhinoviruses 1A, 2, 6, and 13 after 6 or 7 days of incubation. A fifth rhinovirus, 17, formed a 1- to 2-mm feather plaque after 14 days of incubation. Plaque counts of rhinoviruses 1A and 13 were not affected by varying the pH of the overlay from 6.9 to 7.5. Plaque sizes and plaque-forming unit values of high passage rhinoviruses 1A and 13 were equivalent when tested at 26, 31, or 36 C. The rhinoviruses tested were sensitive to incubation at 40 C or heating at 50 C. Enhancement of plaques was observed when Mg(++) was incorporated into agar overlays, but enhancement did not occur when Mg(++) was added to methylcellulose overlays.
描述了一种使用新鲜制备的甲基纤维素覆盖物和人胚二倍体细胞对五种鼻病毒进行灵敏、可靠的蚀斑测定系统。在孵育6或7天后，鼻病毒1A、2、6和13形成了边缘不规则、大小为2毫米的圆形蚀斑。第五种鼻病毒17在孵育14天后形成了1至2毫米的羽毛状蚀斑。将覆盖物的pH值从6.9改变到7.5，对鼻病毒1A和13的蚀斑计数没有影响。当在26、31或36℃下进行测试时，高传代鼻病毒1A和13的蚀斑大小和蚀斑形成单位值相当。所测试的鼻病毒在40℃孵育或50℃加热时敏感。当将Mg(++)加入到琼脂覆盖物中时可观察到蚀斑增强，但当将Mg(++)加入到甲基纤维素覆盖物中时则不会出现蚀斑增强。
