【摘要】 利用表达鸡传染性支气管炎病毒 Gray株 N蛋白的基因工程重组菌 ,表达 IBV - N蛋白 ,并利用表达载体上带有组氨酸标记的特点 ,应用金属螯合填料将其纯化。用纯化的 IBV- N蛋白作为包被抗原 ,成功地建立了一种检测血清中 IBV抗体的间接 EL ISA方法。确定抗原最佳包被浓度为 0 .5 8μg/ ml;被检血清的最佳稀释度为 1∶ 10 0 ;酶标二抗的工作浓度为 1∶ 10 0 0。确定了血清样品阴、阳性临界 OD值为 0 .14。与 IDEXX公司 IBV抗体检测试剂盒比较 ,结果表明自建 EL ISA试剂盒具有较高的敏感性和特异性更多还原

【Abstract】 The recombinant nucleoprotein of the Gray strain of infectious bronchitis virus (IBV) expressed in bacteria with a histidine tag at the amino terminus has been used as antigen for developing an assay to detect IBV-specific antibody. This fusion protein was produced readily in bacteria and easily purified with a nickel column on which bound to the histidine tag.Conditions were optimized for using these preparations for an IBV-specific ELISA. It was shown that the optimal concentration of recombinant N protein for coating of plate was 0.58 μg/ml, the dilution of serum sample was 1∶100, the working concentration of HRP-labeled rabbit anti-chicken IgG was 1∶1 000, Following the determination of conditions of ELISA, the threshold value of ELISA was 0.14. 76 serum samples from chicken were detected for the antibody to IBV by using the developed ELISA and the IDEXX IBV antibody test kit simultaneously. It was found that the specificity and sensitivity of the developed ELISA were very well.更多还原