短波紫外线灭活血小板中病毒的实验研究

An experimental study on the virus inactivation in platelet suspension by using short-wave ultraviolet

目的建立短波紫外线(UVC)对血小板悬液中伪狂犬病毒和辛德毕斯病毒灭活的实验方法,探讨UVC法对血小板计数的影响。方法以伪狂犬病毒和辛德毕斯病毒为试验病毒,Vero细胞为培养细胞,用病毒感染细胞,制备病毒增殖液;用SSP+盐溶液替代血浆重悬血小板;含不同浓度核黄素的血小板SSP+悬液分别以不同辐照剂量的UVC照射,通过细胞感染试验评价灭活效果,筛选最佳灭活条件。用血液细胞分析仪测定UVC处理前后血小板的计数。结果短波紫外线对血小板病毒灭活作用显著,单面辐照强度为4.32 mw/cm2,照射30 s能使血小板SSP+悬液中伪狂犬病毒和辛德毕斯病毒滴度均下降超过4个对数级;照射1 min,伪狂犬病毒全部灭活(下降5.38个对数级),辛德毕斯病毒全部灭活(下降4.59个对数级),UVC处理后血小板计数明显下降,P<0.05;UVC联合核黄素,在相同的照射时间下,添加核黄素组与单独使用UVC组对病毒滴度降低不明显,差异无统计学意义。在辐照剂量相同的条件下,添加核黄素组血小板计数降低较无核黄素组降低缓慢,两两比较差异有统计学意义(P<0.05)。结论 UVC法可有效灭活血小板中伪狂犬病毒和辛德毕斯病毒;SSP+盐溶液替代血浆重悬血小板进行UVC照射,能大大缩短病毒灭活时间,提升灭活效果;UVC联合添加核黄素对病毒灭活没有明显的协同作用,但提示对血小板有一定的保护作用。

Objective To establish a method that can inactivate Pseudorabies and Sindbis virus in platelets by using short wave ultraviolet light C（ UVC） as well as to investigate the effect of UVC on platelet counts. Methods Pseudorabies and Sindbis virus were cultured with Vero cells and platelets were suspended in SSP ＋ solution. Then,the platelet-SSP ＋ suspensions with various concentrations of riboflavin were exposed to different UVC dosage. Finally,the titer of virus was confirmed by the cells infected test. The optimal conditions for inactivation were selected. Additionally,platelet counts were determined by blood cell analyzer after UVC treatment. Results Virus in platelets can be efficiently inactivated by short wave ultraviolet. When using single-sided irradiation intensity at4. 32 mw / cm2 for 30s,the levels of Pseudorabies and Sindbis virus were reduced over 4 log grades. The levels of Pseudorabies virus decreased more than 5. 38 log and Sindbis virus decreased more than 4. 59 log by irradiation for 1 min. Moreover,platelet counts were markedly reduced after UVC treatment（ P〈0. 01）. At the same irradiation dose,the decrease of virus titer in the two groups had no significant difference but the platelet counts of riboflavin group was higher than that of the control group（ P〈0. 05）. Conclusion UVC can inactivate Pseudorabies and Sindbis virus in platelets effectively. Viral inactivation time could be shortened and the effect would be promoted when platelets were resuspended in SSP ＋ solution rather than in plasma. The combination of UVC treatment with riboflavin had no synergistic effect on virus inactivation but had a protective effect on platelet counts.