【摘要】 为建立猪流行性乙型脑炎病毒(JEV)的检测方法,本研究以pr M-E结构蛋白特异性单克隆抗体(MAb)5B10和5E7分别作为捕获抗体和辣根过氧化酶标记的检测抗体,对反应条件进行优化,建立了抗原捕获ELISA(AC-ELISA)方法。该方法与其它常见猪病病原无交叉反应,特异性强。该方法可以检出1.25×105 pfu/m L的病毒和48.44 ng/m L pr M-E重组蛋白,最佳线性检测范围为0.05μg/m L~1.00μg/m L,线性相关系数(R2)为0.996。组装的试剂盒批内和批间重复性试验的变异系数均小于10%,具有良好的重复性。试剂盒4℃保存12个月稳定。该研究为JEV亚单位疫苗的研制与生产提供了技术支持。更多还原

【Abstract】 To develop a method for detecting porcine Japanese encephalitis virus(JEV), an antigen capture ELISA(AC-ELISA) was established with monoclonal antibody(MAb) 5B10 as capture antibody and horseradish peroxidase conjugated MAb 5E7 as detection antibody. The AC-ELISA was specificity which had no cross-reactivity with other related pathogens of swine diseases. The detection limit was 1.25 ×105pfu/m L for JEV and 48.44 ng/m L for viral pr M-E protein expressed by mammalian cell lines. The linear quantitative range was 0.05 μg/m L to 1.00 μg/m L, and the liner correlation(R2) was 0.996. The coefficients of variation intra-assay and inter-assay were less than 10%. The AC-ELISA kit was validity for 12 months stored at 4℃. The established AC-ELISA kit provided technical support to the development and production of JEV subunit vaccine.更多还原