医学毕业维生素c对病毒灭活中血浆蛋白活性的保护效应.docxVIP

XX大学 毕业论文 维生素C对病毒灭活中血浆蛋白活性的保护效应 2014年6月25日 作者：李燕，李明远，蒋仁举，贾文祥 【摘要】 本研究探测在亚甲蓝(methylene blue, MB)光化学法处理单份血 浆过程屮，加入维生素C (vitamineC, VitC)是否影响病毒的灭活效果，是否 对血浆蛋白活性成分具有保护作用。用水泡性口炎病毒(vesicular stomatitis virus, VSV)作为指示病毒，在人血浆中加入不同浓度VitC和终浓度为1 |imol/L的亚 甲蓝，应用40 000 lx荧光强度照射并于不同时间取样检测。以细胞病变效应评 价对VSV的灭活效果，用RT-PCR检测病毒核酸的变化，并采用Clauss法、一 期法和微量免疫电泳等方法对亚甲蓝光化学法处理前后的血浆蛋白含量和活性 进行分析。结果发现，VSV血浆在加入240屮“ol/L Vit C并经MB■光照60分钟 后，病毒滴度下降8 lgTCID50/ml； RT-PCR法也检测不到病毒核酸；血.浆屮 的纤维蛋白原和凝血因了训的回收率分别为83.55%和81.67%,与不加Vit C进 行亚甲蓝光化学法处理结果相比有显著提高(P0.05)；微量免疫电泳显示，血 浆屮的大部分蛋白成分含量没有明显改变，免疫原性也未受到明显影响。结论： 血浆中加入一定量的Vit C不仅不影响亚甲蓝光化学法对病毒的灭活效果，而且 冇效地保护了血浆蛋白活性成分，因此Vit C可作为亚甲蓝光化学法处理血浆时 的保护剂，能有效地提高单份新鲜冷冻血浆的质量。 【关键词】病毒灭活 Protective Effect of Vitamin C on Protien Activity in Plasma during Virus Inactivation Abstract 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 ymol/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 effbet 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 屮nol/L Vit C and treated by MB-light irradiation for 60 min , the titer of VSV decreased by more than 8 lg TICD50/ml.Mean while, target segment amplification of VSV was also negative.The recovery rates of fibrinogen and coagulation factor VIII (FVffl:C ) were 83.55% and 81.67% respectively, which had significant difference compar