PEG-6000纯化流感病毒鸡胚尿囊液的研究  

Study on purification of influenza virus in allantoic fluid of embryonic egg with PEG-6000

作　　者：戴宗祥[1] 耿兴良 周健[1] 马磊[1] 郭晨[1] 周竟贤 高清霞[1] 张新文[1] 姜述德[1] 廖国阳[1] 

机构地区：[1]中国医学科学院/北京协和医学院/医学生物学研究所/云南省重大传染病疫苗研发重点实验室,云南昆明650118

出　　处：《国际检验医学杂志》2014年第1期1-2,5,共3页International Journal of Laboratory Medicine

基　　金：国家国际合作项目基金(2011DFR30420);云南省自然科学基金(2010CD134)

摘　　要：目的探讨PEG沉淀法纯化鸡胚培养流感病毒尿囊液的工艺,对PEG-6000的浓度、沉淀时间、离心力进行优化,为流感病毒抗原和抗体制备提供了便捷的手段。方法向流感病毒鸡胚尿囊液添加PEG后进行高速离心收集沉淀,检测纯化病毒液效价、病毒形态,以及卵清蛋白含量。结果 PEG浓度为4%,沉淀时间2h,离心力8 000g条件下,病毒能完全沉淀,杂蛋白卵清蛋白含量低,病毒颗粒完整,浓缩液中病毒血凝效价为1∶10 240。以纯化病毒免疫山羊3次后,血清抗体滴度为1∶1 024且无交叉。结论本研究初步证明优化后的PEG沉淀法对鸡胚培养流感病毒尿囊液中的病毒浓缩、纯化达到了很好的效果。Objective In this paper, we focus on the procedure of purifying influenza virus and optimizing the concentration of PEG-6000, the concentrating time and centrifugal force. In order to provide a fast way to obtain the influenza virus antigen and anti- body. Methods We centrifuged the influenza virus in allantoic fluid of embryonic egg after adding PEG-6000 to it,then collect the precipitant,and test the contented of ovalbumin ,viral particle integrity,and the titers of virus. Results It showed that when the ex- perimental conditions was setted as 4% for the PEG concentration, 2 hours for the settling time and 8 000 g for the centrifugal force, the virus could be completely precipitated. In this situation, the miscellaneous protein ovalbumin content was low, and the vi- rus was integrate. Finally the hemagglutination titer was 1 ： 10 240,the antiserum titer was 1 = 1 024,and there was no cross. Con- clusion Optimized PEG precipitation method could be with fine effects for the concentration and purification of influenza virus from allantoie fluid of chick embryo.

关 键 词：流感病毒 实验室技术和方法 抗原抗体复合物 

分 类 号：R373.13[医药卫生—病原生物学]