Preston D R, Bitton G, Farrah S R
Department of Microbiology and Cell Science, University of Florida, Gainesville 32611.
Appl Environ Microbiol. 1990 Jan;56(1):295-7. doi: 10.1128/aem.56.1.295-297.1990.
Laboratory strains of enteroviruses, as well as viruses isolated from raw wastewater, were found to exhibit enhanced infectivity in vitro when BGM cell monolayers were pretreated with the cationic polymer polyethyleneimine (PEI). Viruses were assayed by the cytopathic effect technique and as PFU under methylcellulose and agar overlays with monolayers treated with 0 to 5.0 x 10(-3)% (wt/vol) PEI in phosphate-buffered saline supplemented with 2% fetal bovine serum. Poliovirus type 1 cytopathic effect occurred at an enhanced rate in cells treated with 5.0 x 10(-3)% PEI compared with untreated cells. PEI-treated cells were found to adsorb viruses much more effectively than untreated cells did. When the methylcellulose overlay procedure was used, rates of infectivity were enhanced as follows: poliovirus type 1, 5.5-fold; echovirus type 1, 1.2-fold; echovirus type 5, 5.2-fold; and coxsackievirus type B5, 4.9-fold. Viruses concentrated from raw wastewater showed a 3.8-fold increase in titer when quantitated by the most-probable-number method and a 3.3-fold increase when quantitated as PFU under an agar overlay.
当用阳离子聚合物聚乙烯亚胺(PEI)预处理BGM细胞单层时,发现肠道病毒的实验室菌株以及从未经处理的废水中分离出的病毒在体外表现出增强的感染性。病毒通过细胞病变效应技术进行检测,并在含有2%胎牛血清的磷酸盐缓冲盐水中,用0至5.0×10(-3)%(重量/体积)的PEI处理单层细胞,在甲基纤维素和琼脂覆盖下作为空斑形成单位(PFU)进行测定。与未处理的细胞相比,用5.0×10(-3)%的PEI处理的细胞中1型脊髓灰质炎病毒细胞病变效应的发生率更高。发现经PEI处理的细胞比未处理的细胞更有效地吸附病毒。当使用甲基纤维素覆盖程序时,感染率提高如下:1型脊髓灰质炎病毒,5.5倍;1型艾柯病毒,1.2倍;5型艾柯病毒,5.2倍;以及B5型柯萨奇病毒,4.9倍。通过最可能数法对从未经处理的废水中浓缩的病毒进行定量时,其滴度增加了3.8倍,在琼脂覆盖下作为PFU进行定量时,其滴度增加了3.3倍。