Armon R, Arella M, Payment P
Centre de recherche en virologie, Institut Armand Frappier, Laval, Québec, Canada.
Can J Microbiol. 1988 May;34(5):651-5. doi: 10.1139/m88-107.
Addition of Tween 80 to a 1.5% solution of beef extract was found to enhance the elution of bacteriophages adsorbed to electronegative filters. When reconcentration of the eluate was attempted by ammonium sulfate precipitation, a floating layer containing most of the viruses was formed. This floating layer can be obtained with several nonionic detergents including Tween 80 and under a salt saturation of 55% with ammonium sulfate, potassium tartrate, and sodium phosphate. Virus recovery ranged from 91 to 103% and was obtained with several bacteriophage strains. With poliovirus type 1, coxsackievirus B-4, and rotavirus SA-11 the recoveries were 100, 20, and 80%, respectively, but toxicity to cell culture was encountered: after removal of the detergent by a second floating layer method the recovery was 32% for poliovirus. Compared with organic flocculation, this method also had both improved recovery for bacteriophages and protective properties for samples frozen at -70 degrees C.
研究发现，在1.5%的牛肉浸出液溶液中添加吐温80可增强吸附在带负电滤膜上的噬菌体的洗脱效果。当尝试通过硫酸铵沉淀法对洗脱液进行再浓缩时，会形成一个包含大部分病毒的漂浮层。使用包括吐温80在内的几种非离子洗涤剂，并在硫酸铵、酒石酸钾和磷酸钠的盐饱和度为55%的条件下，均可获得该漂浮层。病毒回收率在91%至103%之间，且几种噬菌体菌株均可实现。对于1型脊髓灰质炎病毒、B - 4型柯萨奇病毒和SA - 11型轮状病毒，回收率分别为100%、20%和80%，但对细胞培养存在毒性：通过第二种漂浮层方法去除洗涤剂后，脊髓灰质炎病毒的回收率为32%。与有机絮凝法相比，该方法对噬菌体的回收率更高，且对在-70摄氏度下冷冻的样品具有保护作用。
