EV71抗原ELISA定量检测方法的建立Development of A Quantitative ELISA Method for EV71 Antigen
贾卉;蔡芳;高强;张建三;井申荣;
摘要(Abstract)：
目的建立EV71抗原双抗体夹心ELISA定量检测方法,用于EV71灭活疫苗生产过程中抗原含量的检测。方法采用EV71全病毒免疫,制备抗EV71单克隆抗体及多克隆抗体,以抗EV71多克隆抗体作为包被抗体,经HRP标记的抗EV71单克隆抗体作为酶标抗体,建立双抗体夹心ELISA法,检测样品中EV71抗原含量,并对该方法进行验证及应用。结果所建立的方法线性范围为5～80U/ml,R2=0.994,线性关系良好,定量限度为5U/ml;检测不同浓度的EV71抗原内部参考品,回收率为88%～119.0%;变异系数均小于15%;与甲肝病毒收获液、乙脑病毒灭活液、H5N1流感病毒裂解疫苗原液、小牛血清、人白蛋白、Vero细胞、MEM培养基及CoxA16收获液均无交叉反应;检测EV71灭活疫苗原液纯化过程样品的抗原含量,能有效反映抗原纯化趋势;检测铝吸附疫苗成品解离后抗原含量,准确性较高。结论已建立了EV71抗原双抗体夹心ELISA定量检测方法,该方法线性好,特异性强,灵敏度高,准确性及重复性良好,可用于EV71疫苗生产过程中及终产品的抗原定量检测。
关键词(KeyWords)： EV71抗原;双抗体夹心ELISA;验证
基金项目(Foundation):
作者(Authors): 贾卉;蔡芳;高强;张建三;井申荣;
DOI: 10.13200/j.cjb.2010.01.93.jiah.023
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