Ramia S, Sattar S A
J Clin Microbiol. 1979 Nov;10(5):609-14. doi: 10.1128/jcm.10.5.609-614.1979.
Incorporation of 5 micrograms of trpsin per ml of the overlay (Eagle minimal essential medium-0.7% Ionagar no. 2) was found to be necessary for plaque formation by simian rotavirus SA-11. Plaques of 3 to 4 mm in diameter were produced in MA-104 cells after 5 days of incubation at 37 degrees C. Plaque size was even larger (5 to 6 mm) in monolayers of African green monkey kidney cells. Addition of diethyl-aminotheyl-dextran, protamine sulfate, or 5-bromodeoxyuridine to the trypsin-containing overlay did not improve plaque formation by the virus. Incorporation of beef extract or yeast extract to a final concentration of 0.5% in the trypsin-containing overlay inhibited plaque formation. On the other hand, the presence of lactalbumin hydrolysate or peptone at a similar concentration in the overlay did not inhibit plaque formation. When methylcellulose was used instead of the agar as the solidifying agent in the overlay, no plaques were seen. SA-11 is a useful model for the study of human rotaviruses, and this relatively simple plaque assay system should further enhance its usefulness in this regard.
已发现，每毫升覆盖物（伊格尔最低限度基本培养基 - 0.7% 离子琼脂2号）中加入5微克胰蛋白酶，对于猿猴轮状病毒SA - 11形成噬斑是必要的。在37℃孵育5天后，MA - 104细胞中产生了直径为3至4毫米的噬斑。在非洲绿猴肾细胞单层中，噬斑尺寸甚至更大（5至6毫米）。向含胰蛋白酶的覆盖物中添加二乙氨基乙基葡聚糖、硫酸鱼精蛋白或5 - 溴脱氧尿苷并不能改善病毒的噬斑形成。在含胰蛋白酶的覆盖物中加入牛肉浸出物或酵母浸出物至最终浓度为0.5%会抑制噬斑形成。另一方面，覆盖物中存在类似浓度的乳白蛋白水解物或蛋白胨不会抑制噬斑形成。当在覆盖物中使用甲基纤维素代替琼脂作为固化剂时，未观察到噬斑。SA - 11是研究人类轮状病毒的有用模型，这种相对简单的噬斑测定系统应会进一步提高其在这方面的实用性。
