本文選題：流感病毒 + 實驗室技術和方法��； 參考：《國際檢驗醫(yī)學雜志》2014年01期

【摘要】：目的探討PEG沉淀法純化雞胚培養(yǎng)流感病毒尿囊液的工藝,對PEG-6000的濃度、沉淀時間、離心力進行優(yōu)化,為流感病毒抗原和抗體制備提供了便捷的手段。方法向流感病毒雞胚尿囊液添加PEG后進行高速離心收集沉淀,檢測純化病毒液效價、病毒形態(tài),以及卵清蛋白含量。結果 PEG濃度為4%,沉淀時間2h,離心力8 000g條件下,病毒能完全沉淀,雜蛋白卵清蛋白含量低,病毒顆粒完整,濃縮液中病毒血凝效價為1∶10 240。以純化病毒免疫山羊3次后,血清抗體滴度為1∶1 024且無交叉。結論本研究初步證明優(yōu)化后的PEG沉淀法對雞胚培養(yǎng)流感病毒尿囊液中的病毒濃縮、純化達到了很好的效果。
[Abstract]:Objective to study the purification process of chicken embryo culture influenza virus allantoic fluid by PEG precipitation method, optimize the concentration, precipitation time and centrifugal force of PEG-6000, and provide a convenient method for the preparation of influenza virus antigen and antibody. Methods PEG was added to the chicken embryo allantoic fluid of influenza virus. The purified virus was collected and precipitated by high speed centrifugation. The titer of purified virus, the morphology of the virus and the content of ovalbumin were detected. Results when the concentration of PEG was 4 and the precipitation time was 2 h, the centrifugal force was 8 000 g, the virus could precipitate completely, the content of hybrid protein ovalbumin was low, the virus particles were intact, and the hemagglutination titer of virus in concentrated solution was 1:10 240. After three times of immunization with purified virus, the titer of serum antibody was 1:1 024 and there was no cross-crossing. Conclusion the optimized PEG precipitation method has a good effect on virus concentration and purification in chicken embryo culture influenza virus allantoic fluid.
【作者單位】： 中國醫(yī)學科學院/北京協(xié)和醫(yī)學院/醫(yī)學生物學研究所/云南省重大傳染病疫苗研發(fā)重點實驗室;
【基金】：國家國際合作項目基金(2011DFR30420) 云南省自然科學基金(2010CD134)
【分類號】：R373.13

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本文編號：1802554