【摘要】 目的:构建免疫磁珠检测水中f2噬菌体的方法,为检测水中病毒及相关研究提供基础数据和参考。方法:将兔抗f2噬菌体IgG与微磁珠化学偶联构建免疫磁珠,吸附水中f2噬菌体,然后洗脱免疫磁珠结合的噬菌体,空斑实验检测f2噬菌体的效价。改变洗脱液pH,观察不同pH对免疫磁珠捕获病毒洗脱效果的影响。结果:通过化学方法将抗体包被到磁珠上效果良好,免疫磁珠对f2噬菌体的吸附效率约50%以上,但洗脱效率存在差异,约30%的f2噬菌体可从免疫磁珠上洗脱下来;不同pH值洗脱液洗脱效果有差异,在pH值接近2.7时效果较好。结论:本实验通过化学偶联法成功构建免疫磁珠,并获得了较好的捕获病毒效果,但洗脱解离效率偏低,因此洗脱液及洗脱条件的选择是本研究提高病毒回收率关键因素之一。更多还原

【Abstract】 Objective: To construct the immunomagnetic beads (IMB) to adsorb and separate coliphage f2 in water for establishing a fast and economic method of viral determination.Methods:Immunomagnetic beads were made through coating the antibody to magnetic beads.Bacteriophage f2 in water samples was adsorbed by IMB.Then,eluted the f2 phages from immunomagnetic beads for Plaque assay and observed the elution effects at different pH after being absorbed.Results:Good effect of coating the antibody to magnetic beads has been achieved by using chemical method,so has the adsorption of f2 phages by IMB.But the elution efficiency with which less than 30% bacteriophage f2 was eluted was not desirable;solutions with various pH had different elution effect and the maximum elution values could be obtained at about pH 2.7.Conclusion:In this experiment,construction of IMB by means of chemical coating has a good effect.However,choosing of the eluent plays a key role in improving elution efficiency of virus from IMB.更多还原