Sabin株脊髓灰质炎病毒经透析处理后病毒回收效果评价

Evaluation on recovery of poliovirus Sabin strain after dialysis

目的采用透析法对Sabin株3个型别的脊髓灰质炎病毒收获液进行透析后的病毒回收效果评价。方法高病毒滴度时,对Sabin株Ⅰ、Ⅱ、Ⅲ型病毒收获液分别进行透析,比较每个型别病毒收获液透析前后的病毒感染性滴度回收率,及透析后的甲醛清除率;低病毒滴度时,选取101、100、10-1 CCID_(50)/mL 3个滴度的Sabin株Ⅰ、Ⅱ、Ⅲ型病毒液,分别进行透析,将相同型别及滴度的透析前后样品接种至Hep-2细胞上进行3次传代,比较透析前后细胞病变比率及病毒滴度。结果高病毒滴度时,各型别病毒收获液透析后与透析前相比,样品体积和感染性滴度均未发生较大变化(P>0.05),透析后甲醛清除率达99%以上。101 CCID_(50)/mL低病毒滴度时,各型别病毒3次传代均能100%引起细胞病变;100 CCID_(50)/mL低病毒滴度时,Ⅰ、Ⅱ、Ⅲ型病毒第1次传代引起细胞病变比率分别为66%、33%和33%,第2、3次传代均能100%引起细胞病变;10-1 CCID_(50)/mL低病毒滴度时,3个型别病毒3次传代均不能引起细胞病变。所有细胞病变后检测病毒滴度均在6.0 lgCCID_(50)/mL以上。结论 Sabin株3个型别的病毒收获液经透析后,活病毒的回收率较高,基本无损失,透析法可作为该病毒的纯化方法。

Objective To evaluate the recovery of polio virus Sabin strain of three types after dialysis.Methods Polio virus Sabin strain of types Ⅰ,Ⅱ and Ⅲ,at high titers,were dialyzed.The recoveries of titers of each type before and after dialysis,as well as the clearance rates of formaldehyde after dialysis,were compared.However,the polio virus at titers of 101,100 and 10-1 CCID50/mL,and the samples of the same type and titer before and after dialysis were inoculated to Hep-2 cells and subcultured for three passages.The cytopathic effect（CPE）rates and virus titers before and after dialysis were compared.Results The volume and titer of samples of various types at high titers after dialysis showed no significant difference with those before dialysis（P〈0.05）,while the clearance rate of formaldehyde after dialysis was more than 99%.All the samples of various types at a titer of 101 CCID50/mL caused CPE after subculture for three passages.The CPE rates caused by samples of typesⅠ,Ⅱ and Ⅲ,each at a titer of 100 CCID50/mL,were 66%,33% and 33% respectively after subculture for one passage,all of which were 100% after subculture for two and three passages.However,the samples of various types at a titer of 10-1 CCID50/mL caused no CPE after subculture for three passages.All the virus titers of samples after CPE were more than 6.0 lgCCID50/mL.Conclusion The recovery of three types of polio virus Sabin strain of three types were high after dialysis,with little loss,indicating that dialysis might be used for purification of the virus.