Characterization And Application Of Monoclonal Antibody Against Fusion Protein Of Newcastle Disease Virus

Newcastle disease is defined as an infectious disease of birds caused by a virus of avian paramyxovirus serotype1 (AMPV-1). Since 1997, the goose flocks in South and East of China have suffered severe outbreaks of Newcastle disease virus (NDV) infection. Although measures have been taken to control the disease, it still spreads progressively. In this paper we analyzed mAbs against fusion protein of NDV with different NDV strains isolated from different poultry species and developed a IFA method for detection GNDV infection.CEF infected with NDV F48E8 were prepared for indirect immunoflurescent assay (IFA). The primary hybridoma which secreting monoclonal antibody (mAb) against fusion protein of NDV were subcloned and screened by IFA. Twelve hybridoma cell lines which could secrete the monoclonal antibodies to fusion protein of NDV were obtained, named NDV-F 1B4,NDV-F 4B11,NDV-F5F3,NDV-F 4C1,NDV-F 2H10,NDV-F 6F8,NDV-F 4H10,NDV-F 1D10,NDV-F2F12,NDV-F4D9,NDV-F5C6,NDV-F6C4. The IFA titers of these mAbs were also determined, which were between 1:1600 to 1:12800. Western blot results showed that twelve monoclonal antibodies could recognize 55KD-fusion protein of NDV. Their Enzyme-linked immunosorbent assay (ELISA) reactivity were low or no reaction. 3 of 12 mAbs showed neutralization activity in CEF infected with F48E8 NDV.All twelve mAbs reaction with different NDV strains isolated from different poultry species (such as goose, pigeon, chicken) were analysis by IFA, Western blot and ELISA. The results showed that these mAbs have different reaction with different NDVs in IFA and ELISA. However all of the mAbs showed good reaction with all NDV in Western blot.Based on the reaction mAb recognized all NDVs in IFA, a IFA method for detecting infection of GNDV were established. In the test, the CEF were infected with 100TCID50 GNDV and detected every 3 hours intervals post infection. The result demonstrated that GNDV infection could be detected post infection 18hrs. This mothod is more convenient, quickly and accurately than chicken embryo inoculation and RT-PCR.