【摘要】 以在E coli高效表达的犬冠状病毒核蛋白为抗原,用辣根过氧化物酶(HRP)标记的兔抗犬IgG为二抗,建立了检测犬冠状病毒抗体的间接ELISA方法。经方阵滴定确定出最佳反应条件为1μg 孔纯化的E coli表达的重组N蛋白抗原包被酶标板,用10%的兔血清进行封闭,以正常E coli裂解上清液稀释待检血清。结果表明:应用重组N蛋白作为诊断用抗原具有特异性强、稳定性高、成本较低等特点。更多还原

【Abstract】 An indirect ELISA for detecting antibodies against canine coronavirus was developed using the recombinant nucleoprotein(N) expressed highly in E.coli as antigen and IgG labeled by HRP as secondary. The wells of ELISA plates were coated with 1ug abstracted N protein in 0.05 mol/L carbonate buffer (pH 9.6) and incubated for 2 hours at 37℃,then overnight at 4℃. After washing, 100 μL 10% normal serum of rabbit was added as blocking reagent and incubated for 2 hours at 37℃. Before being added, the serum samples were preincubated with the supernatant of the normal abstracts of E.coli, the non-specific antibodies in the serum samples could be eliminated and it was not necessary to highly purify the antigen. The results showed that the assay is of high specificity, stability and low cost.更多还原