Hill W F, Akin E W, Benton W H, Metcalf T G
Appl Microbiol. 1972 May;23(5):880-8. doi: 10.1128/am.23.5.880-888.1972.
The efficiency of a Millitube MF cartridge filter, a membrane filter, for recovery of poliovirus from 100-gal volumes of both fresh (tap) and estuarine water was determined. In the high multiplicity of virus input-output experiments, recovery of 97% or greater of input virus was achieved in both types of water when the final concentration of divalent cation as Mg(2+) was 1,200 mug/ml and the pH was 4.5. Virus was effectively eluted from the membrane cartridge with 5x nutrient broth in 0.05 M carbonate-bicarbonate buffer at pH 9.0. Four elutions of 250 ml each were used. In the low multiplicity of virus input-output experiments under the same cationic and pH conditions, up to 67% of the input virus was recovered when the virus was further concentrated from the eluates by the aqueous polymer two-phase separation technique. The volume reduction was 126,000-190,000 to 1. The use of the combined techniques, i.e., membrane adsorption followed by aqueous polymer two-phase separation, provided a highly sensitive, simple, and remarkably reliable sequential methodology for the quantitative recovery of poliovirus occurring at multiplicities as low as 1 to 2 plaque-forming units per 5 gal of water.
测定了Millitube MF筒式过滤器(一种膜过滤器)从100加仑新鲜(自来水)和河口水中回收脊髓灰质炎病毒的效率。在高病毒输入-输出复数实验中,当二价阳离子Mg(2+)的终浓度为1200微克/毫升且pH值为4.5时,两种类型的水中均实现了97%或更高的输入病毒回收率。在pH值为9.0的0.05 M碳酸盐-碳酸氢盐缓冲液中,用5倍营养肉汤可有效地从膜筒中洗脱病毒。每次进行4次250毫升的洗脱。在相同阳离子和pH条件下的低病毒输入-输出复数实验中,当通过水性聚合物双相分离技术从洗脱液中进一步浓缩病毒时,可回收高达67%的输入病毒。体积减少至126,000 - 190,000分之一。联合使用这些技术,即膜吸附后进行水性聚合物双相分离,为定量回收脊髓灰质炎病毒提供了一种高度灵敏、简单且非常可靠的连续方法,该病毒在每5加仑水中的复数低至1至2个空斑形成单位。