核黄素光化学法灭活Sindbis病毒的研究

Research of the inactivation efficiency of riboflavin photochemical on Sindbis virus

目的探究核黄素光化学反应对Sindbis病毒的灭活效果。方法将10 mmol/L的核黄素0.050 mL加入到4.95 mL的Sindbis病毒(XJ-160株)悬液中,经440 nm波段的可见光(40 J/cm2)双侧照射后,接种至幼仓鼠肾细胞(BHK-21)中培养,观察细胞病变情况(CPE),检测病毒滴度,用PCR检测病毒核酸的变化,并在透射电镜下观察病毒形态。结果经终浓度为100μmol/L核黄素结合440 nm可见光作用,可将滴度为6.5 log TCID50Sindbis病毒灭活至≤0.5 logTCID50;PCR法未能扩增出病毒核酸片段;透射电镜显示病毒颗粒塌陷。结论核黄素光化学法能有效灭活Sindbis病毒,其主要作用靶点为核酸。

Objective To study the inactivation efficiency of riboflavin photochemical on Sindbis virus in this research,and explore its mechanisms.Methods The 0.050 mL riboflavin with a concentration of 10 mmol/L was added into 4.95 mL sindbis suspension,and then exposed to visible light(440 nm) at 40 J/cm2.The effect and the mechanism of Sindbis virus was analyzed by BHK cells cultivation,transmission electron technology and PCR.Results When concentration of riboflavin was 100 μmol/L,and illumination of visible light(440 nm),the titter of Sindbis virus was reduced from 6.5 logTCID50 to≤0.5 logTCID50.The segment of nucleic acid of Sindbis virus couldn′t be amplified with PCR,and transmission electron photology showed that the identification of Sindbis virions was collapsed.Conclusion Riboflavin photochemical can effectively inactivate Sindbis virus and damage their nuclear acid.