不同储存条件和反复冻融次数对口蹄疫病毒滴度的影响

Effects of different storage conditions and repeated freeze-thaw cycles on the titer of Foot-and-Mouth disease virus

为研究不同生物学条件下,口蹄疫病毒感染BHK-21细胞后病毒滴度的影响,试验将收集O、A 2种亚型的口蹄疫病毒液,经过不同次数的反复冻融(0、1、2、3、4、5、6次),设定不同的温度条件(20、-4、-20、-80℃),间隔不同的保存时间(0、7、14、21、28 d),测定其半数组织细胞感染量(TCID 50),以计算病毒滴度。结果表明:O型、A型口蹄疫病毒的细胞培养液分别反复冻融4次、3次后,其病毒滴度最高;相同存放温度下,随着保存时间的不断延长,病毒滴度表现略微下降的趋势;在一定保存时间内,随着存放温度不断降低,病毒滴度有所改变,其最适存放温度为-20℃和-80℃。因此,适宜的冻融次数、保存温度及存放时间对保存2种亚型的口蹄疫病毒具有一定的实际意义,为病毒的实验室应用提供参考依据。

In order to study the effect of virus titer on BHK-21 cells infected by Foot-and-Mouth disease virus under different biological conditions,we collected the Foot-and-Mouth disease virus liquids of O and A subtypes,after repeated freeze-thaw cycles(0,1,2,3,4,5,6 times),set different temperature conditions(20℃,-4℃,-20℃,-80℃)and different storage time(0,7,14,21,28 d),then half of the tissue cell infection amount(TCID 50)was measured to calculate the virus titer.The results showed that the virus titers were the highest when the cell culture medium containing O-type and A-type Foot-and-Mouth disease virus was repeatedly frozen and thawed 4 times and 3 times,respectively.At the same storage temperature,the virus titer showed a slight decline with the increase of the storage time.In a certain storage time,the virus titer changed with the decrease of storage temperature,and the optimal storage temperature was-20℃and-80℃.Therefore,the appropriate freeze-thaw cycles,storage temperature and storage time have certain reference functions for preserving the two subtypes of Foot-and-Mouth disease virus,and it provides a reference for laboratory application of virus.