摘要
目的 建立一种简便、有效的从细胞培养物中纯化PV1病毒的方法用于水病毒消毒实验 .方法 PV 1 Henan株在 Hep- 2细胞中增殖后用聚乙二醇 (PEG) 6 0 0 0沉淀结合差速离心的方法纯化病毒 .RT- PCR和透射电镜 (TEM)鉴定提纯病毒的形态学和分子生物学特性 .结果 p H值 7.5时终浓度 70 g· L- 1的 PEG6 0 0 0沉淀病毒的感染性回收率(PFU计数 )为 79.2 % ,提纯系数 (PF)为 1 0 6 ,再经差速离心(1 6 0 0 0 g30 m in,1 0 0 0 0 0 g4.5 h)浓缩病毒的感染性回性率和提纯系数分别为 2 9.0 %和 1 6 8.RT- PCR和 TEM最终鉴定提纯物为 PV 1 Henan株 .结论 PEG 6 0 0 0结合差速离心的方法较之蔗糖密度梯度区带离心方法易于操作 ,且比单独PEG沉淀有更高的纯化系数 ,是一种简便、有效的方法 .
AIM To establish a convenient and effective method for the isolation and purification of polioviruses. METHODS PV1 Henan strain propagated in Hep 2 cells was precipitated by PEG 6000 in combination with differential centrifugation. RESULTS 7% final concentration of PEG 6000 at a pH value of 7.5 was found to give a 79.2% recovery of virus infectivity and the purification factor (PF) was 106. The recovery and PF with differential centrifugation were 29.0% and 168. Transmission electron microscopy (TEM) and RT PCR determined viral morphologic and molecular biological characterization of the final preparation. CONCLUSION This method is easier to operate than sucrose density gradient rate zonal centrifugation and has a higher PF than that of PEG 6000precipitation alone.
出处
《第四军医大学学报》
2000年第1期38-40,共3页
Journal of the Fourth Military Medical University
