IB-ELISA诊断试剂的研究

目的　为鸡群日常监测传染性支气管炎病毒 (IBV)抗体提供一个方便、特异、快速的检测方法。方法　利用 9日龄SPF鸡胚尿囊腔接种培养IBV ,以及鸡胚成纤维细胞传代培养IBV ,经纯化后 ,测定蛋白含量 ,作为包被抗原 ,利用过碘酸盐法标记兔抗鸡IgG作为酶结合物 ,用TMB作为显色剂 ,建立起酶联免疫吸附试验 (ELISA)技术 ,检测血清样品中传染性支气管炎抗体。结果　上述两种方法制备的抗原皆可用于此试验 ,与NDV、MDV、IBDV、AIV、APIV、REV、ILTV、IC、MG、MS等抗血清无交叉反应 ,与IDEXXIB ELISA比较 ,结果吻合较好。结论　本研究制备的IB ELISA诊断试剂 ,可用于日常监测传染性支气管炎病毒抗体

Study on IB-ELISA Diagnostic Reagent

Purpose To establish a convenient,specific and rapid detective technique for routine measuring antibodies to infectious bronchitis virus.Method IBV was propagated in the allantoic cavity of 9-day-old specific pathogen free(SPF),as well as in the material of chicken embryo fibroblasts(CEF)by serial passage.Followed purification,the protein quantity was measured,which was used as coated antigen.Conjugation was labeled rabbit anti-chichen IgG by using periodate method,TMB and OPD as substrate,to set up enzyme-linked immunosorbent assay(ELISA)for testing infectious bronchitis virus antibodies.Results The above two obtained IBV could be used as coating antigen.No crossing reaction was observed with antibodies to Newcastle disease virus(NDV),Marek's disease virus(MDV),infectious bursal disease virus(IBDV),infectious laryngotracheitis virus(ILTV),Avian influenza virus(AIV),Avian parainfluenza virus(APIV),Reticuloendotheliosis virus(REV),Infectious Coryza(IC),Mycoplasma gallisepticum(MG),Mycoplasma synoviae(MS).The coincidence of result is good by comparing with the imported reagent of IB-ELISA produced by IDEXX.Conclusion:The IB-ELISA diagnostic reagent can be used to detect IBV-antibody.