鸡传染性支气管炎ELISA抗体检测试剂盒的研制

Study on IBV ELISA Kit for Detection of Infectious Bronchitis Virus Specific Antibodies

采用加蔗糖垫离心纯化鸡传染性支气管炎病毒(IBV)制备抗原 ,用提纯的IBV抗原包被微量板,建立了检测鸡传染性支气管炎 (IB)抗体的ELISA试剂盒。抗原、被检血清和酶标结合物的最佳工作浓度分别为10ug/ml、1∶200和1∶3200。与IDEXX试剂盒相比 ,其敏感性、重复性、特异性均接近国外同类产品水平。对SPF鸡血清、实验免疫与攻毒的SPF鸡血清进行检测 ,结果表明所建立的ELISA特异性为95.6% ,与IDEXX试剂盒符合率为95.6%。用于检测抗IBV特异性IgG抗体发现在免疫接种IB弱毒苗后 ,第4天即可检测到IgG抗体 ,峰值在第3周。试验鸡在通过滴鼻、点眼途径人工感染IBV强毒后 ,第5天抗体滴度明显上升。我们认为 ,该法是目前我国SPF鸡质量监测、养鸡生产中进行IB监测较好的方法。

An ELISA kit for detecting specific IgG against Infectious Bronchitis.Virus(IBV) was purified by sucrose cushion centrifuge established . The optimum concentration of coating antigen was 10 ug/ml,and the sera dilution 1:200 and the conjugate dilution 1:3200. The results of the new IBV ELISA kit showed that this assay was useful for diagnosing IBV infection. The sensitivity,repetition and specificity of the method are as good as that of the similar foreign products. By detecting the sera from SPF, immunized and challenged chickens, the specificity of the IBV-IgG ELISA was 95.6%,which corresponed 95.6% with that of IDEXX IBV-ELISA kit. The method could detect the IgG to IBV in 4 days after chicken vaccinated with IBV strain H52 and the titer of IgG reached highest in 3 weeks. Antibody titer developed remarkablely after infected with virulent IBV-M41 strain via the oculonasal and eye routes .