Li Yan, Li Ming-Yuan, Jiang Ren-Ju, Jia Wen-Xiang
Departmnet of Microbiology, West China College of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2006 Apr;14(2):392-6.
To determine whether addition of vitamin C (Vit C) to single-unit plasma could influence the efficacy of inactivating viruses and could maintain the activity of plasma proteins by methylene blue (MB)-light treatment. Vesicular stomatitis virus (VSV) Indiana strain was used as the indicating virus. Human plasma containing VSV was added with different concentrations of Vit C and final concentration 1 micromol/L MB and irradiated by fluorescence at an intensity of 40,000 lx, samples were collected at different times for detection. Cytopathic effect was used to test the effect of virus inactivation. A segment of the nucleic acid encoding capsid protein of VSV was amplified with RT-PCR. Some methods, such as the Clauss method, the one-stage method, microimmunoelectrophoresis, were used to investigate the changes of plasma components. The results showed that when the VSV plasma was added with 240 micromol/L Vit C and treated by MB-light irradiation for 60 min, the titer of VSV decreased by more than 8 lg TICD50/ml. Meanwhile, target segment amplification of VSV was also negative. The recovery rates of fibrinogen and coagulation factor VIII (FVIII: C) were 83.55% and 81.67% respectively, which had significant difference comparing with the routine MB-fluorescent light treatment. Most of plasma proteins were not affected significantly. No change in immunogenicity of these proteins was observed by using microimmunoelectrophoresis. It is concluded that virus inactivation is not influenced and plasma proteins are effectively protected by Vit C. Vit C can be used as a protector and is beneficial to improving the quality of plasma subjected to MB- photodynamic treatment.
为确定在单单位血浆中添加维生素C（Vit C）是否会影响病毒灭活效果，并通过亚甲蓝（MB）光处理维持血浆蛋白活性。使用水疱性口炎病毒（VSV）印第安纳株作为指示病毒。向含有VSV的人血浆中添加不同浓度的Vit C和终浓度为1 μmol/L的MB，并以40,000 lx的强度进行荧光照射，在不同时间收集样本进行检测。采用细胞病变效应检测病毒灭活效果。用RT-PCR扩增VSV衣壳蛋白编码核酸片段。采用克劳斯法、一步法、微量免疫电泳等方法研究血浆成分变化。结果显示，当VSV血浆中添加240 μmol/L Vit C并经MB光照射60分钟后，VSV滴度下降超过8 lg TICD50/ml。同时，VSV的靶片段扩增也为阴性。纤维蛋白原和凝血因子VIII（FVIII:C）的回收率分别为83.55%和81.67%，与常规MB荧光光处理相比有显著差异。大多数血浆蛋白未受到明显影响。通过微量免疫电泳观察这些蛋白的免疫原性无变化。结论是，Vit C不影响病毒灭活，且能有效保护血浆蛋白。Vit C可作为保护剂，有利于提高经MB光动力处理的血浆质量。
